RNA interference mediated inhibition of histone deacetylase (HDAC) gene expression using short interfering nucleic acid (siNA)

FIELD OF THE INVENTION

The present invention relates to compounds, compositions, and methods for the study, diagnosis, and treatment of traits, diseases and conditions that respond to the modulation of histone deacetylase (HDAC) gene expression and/or activity. The present invention is also directed to compounds, compositions, and methods relating to traits, diseases and conditions that respond to the modulation of expression and/or activity of genes involved in HDAC gene expression pathways or other cellular processes that mediate the maintenance or development of such traits, diseases and conditions. Specifically, the invention relates to small nucleic acid molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA) molecules capable of mediating or that mediate RNA interference (RNAi) against gene expression. Such small nucleic acid molecules are useful, for example, in providing compositions for treatment of traits, diseases and conditions that can respond to modulation of HDAC gene expression in a subject or organism, such as cancer and other proliferative diseases or conditions that are associated with HDAC gene expression or activity.

BACKGROUND OF THE INVENTION

The following is a discussion of relevant art pertaining to RNAi. The discussion is provided only for understanding of the invention that follows. The summary is not an admission that any of the work described below is prior art to the claimed invention.

RNA interference refers to the process of sequence-specific post-transcriptional gene silencing in animals mediated by short interfering RNAs (siRNAs) (Zamore et al., 2000, Cell, 101, 25-33; Fire et al., 1998, Nature, 391, 806; Hamilton et al., 1999, Science, 286, 950-951; Lin et al., 1999, Nature, 402, 128-129; Sharp, 1999, Genes & Dev., 13:139-141; and Strauss, 1999, Science, 286, 886). The corresponding process in plants (Heifetz et al., International PCT Publication No. WO 99/61631) is commonly referred to as post-transcriptional gene silencing or RNA silencing and is also referred to as quelling in fungi. The process of post-transcriptional gene silencing is thought to be an evolutionarily-conserved cellular defense mechanism used to prevent the expression of foreign genes and is commonly shared by diverse flora and phyla (Fire et al., 1999, Trends Genet., 15, 358). Such protection from foreign gene expression may have evolved in response to the production of double-stranded RNAs (dsRNAs) derived from viral infection or from the random integration of transposon elements into a host genome via a cellular response that specifically destroys homologous single-stranded RNA or viral genomic RNA. The presence of dsRNA in cells triggers the RNAi response through a mechanism that has yet to be fully characterized. This mechanism appears to be different from other known mechanisms involving double stranded RNA-specific ribonucleases, such as the interferon response that results from dsRNA-mediated activation of protein kinase PKR and 2′,5′-oligoadenylate synthetase resulting in non-specific cleavage of mRNA by ribonuclease L (see for example U.S. Pat. Nos. 6,107,094; 5,898,031; Clemens et al., 1997, J. Interferon & Cytokine Res., 17, 503-524; Adah et al., 2001, Curr. Med. Chem., 8, 1189).

The presence of long dsRNAs in cells stimulates the activity of a ribonuclease III enzyme referred to as dicer (Bass, 2000, Cell, 101, 235; Zamore et al., 2000, Cell, 101, 25-33; Hammond et al., 2000, Nature, 404, 293). Dicer is involved in the processing of the dsRNA into short pieces of dsRNA known as short interfering RNAs (siRNAs) (Zamore et al., 2000, Cell, 101, 25-33; Bass, 2000, Cell, 101, 235; Berstein et al., 2001, Nature, 409, 363). Short interfering RNAs derived from dicer activity are typically about 21 to about 23 nucleotides in length and comprise about 19 base pair duplexes (Zamore et al., 2000, Cell, 101, 25-33; Elbashir et al., 2001, Genes Dev., 15, 188). Dicer has also been implicated in the excision of 21- and 22-nucleotide small temporal RNAs (stRNAs) from precursor RNA of conserved structure that are implicated in translational control (Hutvagner et al., 2001, Science, 293, 834). The RNAi response also features an endonuclease complex, commonly referred to as an RNA-induced silencing complex (RISC), which mediates cleavage of single-stranded RNA having sequence complementary to the antisense strand of the siRNA duplex. Cleavage of the target RNA takes place in the middle of the region complementary to the antisense strand of the siRNA duplex (Elbashir et al., 2001, Genes Dev., 15, 188).

RNAi has been studied in a variety of systems. Fire et al., 1998, Nature, 391, 806, were the first to observe RNAi in C. elegans. Bahramian and Zarbl, 1999, Molecular and Cellular Biology, 19, 274-283 and Wianny and Goetz, 1999, Nature Cell Biol., 2, 70, describe RNAi mediated by dsRNA in mammalian systems. Hammond et al., 2000, Nature, 404, 293, describe RNAi in Drosophila cells transfected with dsRNA. Elbashir et al., 2001, Nature, 411, 494 and Tuschl et al., International PCT Publication No. WO 01/75164, describe RNAi induced by introduction of duplexes of synthetic 21-nucleotide RNAs in cultured mammalian cells including human embryonic kidney and HeLa cells. Recent work in Drosophila embryonic lysates (Elbashir et al., 2001, EMBO J., 20, 6877 and Tuschl et al., International PCT Publication No. WO 01/75164) has revealed certain requirements for siRNA length, structure, chemical composition, and sequence that are essential to mediate efficient RNAi activity. These studies have shown that 21-nucleotide siRNA duplexes are most active when containing 3′-terminal dinucleotide overhangs. Furthermore, complete substitution of one or both siRNA strands with 2′-deoxy (2′-H) or 2′-O-methyl nucleotides abolishes RNAi activity, whereas substitution of the 3′-terminal siRNA overhang nucleotides with 2′-deoxy nucleotides (2′-H) was shown to be tolerated. Single mismatch sequences in the center of the siRNA duplex were also shown to abolish RNAi activity. In addition, these studies also indicate that the position of the cleavage site in the target RNA is defined by the 5′-end of the siRNA guide sequence rather than the 3′-end of the guide sequence (Elbashir et al., 2001, EMBO J., 20, 6877). Other studies have indicated that a 5′-phosphate on the target-complementary strand of a siRNA duplex is required for siRNA activity and that ATP is utilized to maintain the 5′-phosphate moiety on the siRNA (Nykanen et al., 2001, Cell, 107, 309).

Studies have shown that replacing the 3′-terminal nucleotide overhanging segments of a 21-mer siRNA duplex having two-nucleotide 3′-overhangs with deoxyribonucleotides does not have an adverse effect on RNAi activity. Replacing up to four nucleotides on each end of the siRNA with deoxyribonucleotides has been reported to be well tolerated, whereas complete substitution with deoxyribonucleotides results in no RNAi activity (Elbashir et al., 2001, EMBO J., 20, 6877 and Tuschl et al., International PCT Publication No. WO 01/75164). In addition, Elbashir et al., supra, also report that substitution of siRNA with 2′-O-methyl nucleotides completely abolishes RNAi activity. Li et al., International PCT Publication No. WO 00/44914, and Beach et al., International PCT Publication No. WO 01/68836 preliminarily suggest that siRNA may include modifications to either the phosphate-sugar backbone or the nucleoside to include at least one of a nitrogen or sulfur heteroatom, however, neither application postulates to what extent such modifications would be tolerated in siRNA molecules, nor provides any further guidance or examples of such modified siRNA. Kreutzer et al., Canadian Patent Application No. 2,359,180, also describe certain chemical modifications for use in dsRNA constructs in order to counteract activation of double-stranded RNA-dependent protein kinase PKR, specifically 2′-amino or 2′-O-methyl nucleotides, and nucleotides containing a 2′-O or 4′-C methylene bridge. However, Kreutzer et al. similarly fails to provide examples or guidance as to what extent these modifications would be tolerated in dsRNA molecules.

Parrish et al., 2000, Molecular Cell, 6, 1077-1087, tested certain chemical modifications targeting the unc-22 gene in C. elegans using long (>25 nt) siRNA transcripts. The authors describe the introduction of thiophosphate residues into these siRNA transcripts by incorporating thiophosphate nucleotide analogs with T7 and T3 RNA polymerase and observed that RNAs with two phosphorothioate modified bases also had substantial decreases in effectiveness as RNAi. Further, Parrish et al. reported that phosphorothioate modification of more than two residues greatly destabilized the RNAs in vitro such that interference activities could not be assayed. Id. at 1081. The authors also tested certain modifications at the 2′-position of the nucleotide sugar in the long siRNA transcripts and found that substituting deoxynucleotides for ribonucleotides produced a substantial decrease in interference activity, especially in the case of Uridine to Thymidine and/or Cytidine to deoxy-Cytidine substitutions. Id. In addition, the authors tested certain base modifications, including substituting, in sense and antisense strands of the siRNA, 4-thiouracil, 5-bromouracil, 5-iodouracil, and 3-(aminoallyl)uracil for uracil, and inosine for guanosine. Whereas 4-thiouracil and 5-bromouracil substitution appeared to be tolerated, Parrish reported that inosine produced a substantial decrease in interference activity when incorporated in either strand. Parrish also reported that incorporation of 5-iodouracil and 3-(aminoallyl)uracil in the antisense strand resulted in a substantial decrease in RNAi activity as well.

The use of longer dsRNA has been described. For example, Beach et al., International PCT Publication No. WO 01/68836, describes specific methods for attenuating gene expression using endogenously-derived dsRNA. Tuschl et al., International PCT Publication No. WO 01/75164, describe a Drosophila in vitro RNAi system and the use of specific siRNA molecules for certain functional genomic and certain therapeutic applications; although Tuschl, 2001, Chem. Biochem., 2, 239-245, doubts that RNAi can be used to cure genetic diseases or viral infection due to the danger of activating interferon response. Li et al., International PCT Publication No. WO 00/44914, describe the use of specific long (141 bp-488 bp) enzymatically synthesized or vector expressed dsRNAs for attenuating the expression of certain target genes. Zernicka-Goetz et al., International PCT Publication No. WO 01/36646, describe certain methods for inhibiting the expression of particular genes in mammalian cells using certain long (550 bp-714 bp), enzymatically synthesized or vector expressed dsRNA molecules. Fire et al., International PCT Publication No. WO 99/32619, describe particular methods for introducing certain long dsRNA molecules into cells for use in inhibiting gene expression in nematodes. Plaetinck et al., International PCT Publication No. WO 00/01846, describe certain methods for identifying specific genes responsible for conferring a particular phenotype in a cell using specific long dsRNA molecules. Mello et al., International PCT Publication No. WO 01/29058, describe the identification of specific genes involved in dsRNA-mediated RNAi. Pachuck et al., International PCT Publication No. WO 00/63364, describe certain long (at least 200 nucleotide) dsRNA constructs. Deschamps Depaillette et al., International PCT Publication No. WO 99/07409, describe specific compositions consisting of particular dsRNA molecules combined with certain anti-viral agents. Waterhouse et al., International PCT Publication No. 99/53050 and 1998, PNAS, 95, 13959-13964, describe certain methods for decreasing the phenotypic expression of a nucleic acid in plant cells using certain dsRNAs. Driscoll et al., International PCT Publication No. WO 01/49844, describe specific DNA expression constructs for use in facilitating gene silencing in targeted organisms.

Others have reported on various RNAi and gene-silencing systems. For example, Parrish et al., 2000, Molecular Cell, 6, 1077-1087, describe specific chemically-modified dsRNA constructs targeting the unc-22 gene of C. elegans. Grossniklaus, International PCT Publication No. WO 01/38551, describes certain methods for regulating polycomb gene expression in plants using certain dsRNAs. Churikov et al., International PCT Publication No. WO 01/42443, describe certain methods for modifying genetic characteristics of an organism using certain dsRNAs. Cogoni et al, International PCT Publication No. WO 01/53475, describe certain methods for isolating a Neurospora silencing gene and uses thereof. Reed et al., International PCT Publication No. WO 01/68836, describe certain methods for gene silencing in plants. Honer et al., International PCT Publication No. WO 01/70944, describe certain methods of drug screening using transgenic nematodes as Parkinson's Disease models using certain dsRNAs. Deak et al., International PCT Publication No. WO 01/72774, describe certain Drosophila-derived gene products that may be related to RNAi in Drosophila. Arndt et al., International PCT Publication No. WO 01/92513 describe certain methods for mediating gene suppression by using factors that enhance RNAi. Tuschl et al., International PCT Publication No. WO 02/44321, describe certain synthetic siRNA constructs. Pachuk et al., International PCT Publication No. WO 00/63364, and Satishchandran et al., International PCT Publication No. WO 01/04313, describe certain methods and compositions for inhibiting the function of certain polynucleotide sequences using certain long (over 250 bp), vector expressed dsRNAs. Echeverri et al., International PCT Publication No. WO 02/38805, describe certain C. elegans genes identified via RNAi. Kreutzer et al., International PCT Publications Nos. WO 02/055692, WO 02/055693, and EP 1144623 B1 describes certain methods for inhibiting gene expression using dsRNA. Graham et al., International PCT Publications Nos. WO 99/49029 and WO 01/70949, and AU 4037501 describe certain vector expressed siRNA molecules. Fire et al., U.S. Pat. No. 6,506,559, describe certain methods for inhibiting gene expression in vitro using certain long dsRNA (299 bp-1033 bp) constructs that mediate RNAi. Martinez et al., 2002, Cell, 110, 563-574, describe certain single stranded siRNA constructs, including certain 5′-phosphorylated single stranded siRNAs that mediate RNA interference in Hela cells. Harborth et al., 2003, Antisense & Nucleic Acid Drug Development, 13, 83-105, describe certain chemically and structurally modified siRNA molecules. Chiu and Rana, 2003, RNA, 9, 1034-1048, describe certain chemically and structurally modified siRNA molecules. Woolf et al., International PCT Publication Nos. WO 03/064626 and WO 03/064625 describe certain chemically modified dsRNA constructs. Hornung et al., 2005, Nature Medicine, 11, 263-270, describe the sequence-specific potent induction of IFN-alpha by short interfering RNA in plasmacytoid dendritic cells through TLR7. Judge et al., 2005, Nature Biotechnology, Published online: 20 Mar. 2005, describe the sequence-dependent stimulation of the mammalian innate immune response by synthetic siRNA. Yuki et al., International PCT Publication Nos. WO 05/049821 and WO 04/048566, describe certain methods for designing short interfering RNA sequences and certain short interfering RNA sequences with optimized activity. Saigo et al., US Patent Application Publication No. US20040539332, describe certain methods of designing oligo- or polynucleotide sequences, including short interfering RNA sequences, for achieving RNA interference. Tei et al., International PCT Publication No. WO 03/044188, describe certain methods for inhibiting expression of a target gene, which comprises transfecting a cell, tissue, or individual organism with a double-stranded polynucleotide comprising DNA and RNA having a substantially identical nucleotide sequence with at least a partial nucleotide sequence of the target gene. Curtin and Glaser, 2003, Curr. Med. Chem., 10, 2372-92, describe certain siRNAs targeting HDACs. Filocamo et al., International PCT Publication No. WO 05/071079, describe certain siRNA molecules targeting HDAC 11.

SUMMARY OF THE INVENTION

This invention relates to compounds, compositions, and methods useful for modulating histone deacetylase (HCAC) gene expression using short interfering nucleic acid (siNA) molecules. This invention also relates to compounds, compositions, and methods useful for modulating the expression and activity of other genes involved in pathways of HDAC gene expression and/or activity by RNA interference (RNAi) using small nucleic acid molecules. In particular, the instant invention features small nucleic acid molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA) molecules and methods used to modulate the expression of HDAC genes.

A siNA of the invention can be unmodified or chemically-modified. A siNA of the instant invention can be chemically synthesized, expressed from a vector or enzymatically synthesized. The instant invention also features various chemically-modified synthetic short interfering nucleic acid (siNA) molecules capable of modulating HDAC gene expression or activity in cells by RNA interference (RNAi). The use of chemically-modified siNA improves various properties of native siNA molecules through increased resistance to nuclease degradation in vivo and/or through improved cellular uptake. Further, contrary to earlier published studies, siNA having multiple chemical modifications retains its RNAi activity. The siNA molecules of the instant invention provide useful reagents and methods for a variety of therapeutic, cosmetic, veterinary, diagnostic, target validation, genomic discovery, genetic engineering, and pharmacogenomic applications.

In one embodiment, the invention features one or more siNA molecules and methods that independently or in combination modulate the expression of HDAC genes encoding proteins, such as HDAC proteins that are associated with the maintenance and/or development of cancer or proliferative diseases or conditions in a subject or organism, including genes encoding sequences comprising those sequences referred to by GenBank Accession Nos. shown in Table I, referred to herein generally as HDAC. The description below of the various aspects and embodiments of the invention is provided with reference to exemplary HDAC genes (e.g., HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7). Such genes are involved in histone deacetylase activity and associaated epigenetic transcriptional silencing activity via maintenance of heterochromatin (see for example Acharya et al., 2005, Molecular Pharmacology Fast Forward, June 14, 1-49). However, the various aspects and embodiments are also directed to other histone deacetylase genes, such as HDAC homolog genes and transcript variants and polymorphisms (e.g., single nucleotide polymorphism, (SNPs)) associated with certain HDAC genes. As such, the various aspects and embodiments are also directed to other genes that are involved in HDAC mediated pathways of signal transduction or gene expression that are involved, for example, in the maintenance and/or development of conditions or disease states such as cancer and proliferative disease in a subject or organism. These additional genes can be analyzed for target sites using the methods described for HDAC genes herein. Thus, the modulation of other genes and the effects of such modulation of the other genes can be performed, determined, and measured as described herein.

In one embodiment, the invention features a double stranded nucleic acid molecule, such as an siNA molecule, where one of the strands comprises nucleotide sequence having complementarity to a predetermined HDAC nucleotide sequence in a target HDAC nucleic acid molecule, or a portion thereof. In one embodiment, the predetermined HDAC nucleotide sequence is a HDAC nucleotide target sequence described herein. In another embodiment, the predetermined HDAC nucleotide sequence is a HDAC target sequence as is known in the art.

In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a target HDAC RNA, wherein said siNA molecule comprises about 15 to about 28 base pairs.

In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a target HDAC RNA via RNA interference (RNAi), wherein the double stranded siNA molecule comprises a first and a second strand, each strand of the siNA molecule is about 15 to about 30 nucleotides in length, the first strand of the siNA molecule comprises nucleotide sequence having sufficient complementarity to the target HDAC RNA for the siNA molecule to direct cleavage of the target HDAC RNA via RNA interference, and the second strand of said siNA molecule comprises nucleotide sequence that is complementary to the first strand.

In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a target HDAC RNA via RNA interference (RNAi), wherein the double stranded siNA molecule comprises a first and a second strand, each strand of the siNA molecule is about 18 to about 23 nucleotides in length, the first strand of the siNA molecule comprises nucleotide sequence having sufficient complementarity to the target HDAC RNA for the siNA molecule to direct cleavage of the target HDAC RNA via RNA interference, and the second strand of said siNA molecule comprises nucleotide sequence that is complementary to the first strand.

In one embodiment, the invention features a chemically synthesized double stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a target HDAC RNA via RNA interference (RNAi), wherein each strand of the siNA molecule is about 15 to about 30 nucleotides in length; and one strand of the siNA molecule comprises nucleotide sequence having sufficient complementarity to the target HDAC RNA for the siNA molecule to direct cleavage of the target HDAC RNA via RNA interference.

In one embodiment, the invention features a chemically synthesized double stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a target HDAC RNA via RNA interference (RNAi), wherein each strand of the siNA molecule is about 18 to about 23 nucleotides in length; and one strand of the siNA molecule comprises nucleotide sequence having sufficient complementarity to the target HDAC RNA for the siNA molecule to direct cleavage of the target RNA via RNA interference.

In one embodiment, the invention features a siNA molecule that down-regulates expression of a target HDAC gene or that directs cleavage of a target HDAC RNA, for example, wherein the target HDAC gene or RNA comprises protein encoding sequence. In one embodiment, the invention features a siNA molecule that down-regulates expression of a target HDAC gene or that directs cleavage of a target HDAC RNA, for example, wherein the target HDAC gene or RNA comprises non-coding sequence or regulatory elements involved in target HDAC gene expression (e.g., non-coding RNA).

In one embodiment, a siNA of the invention is used to inhibit the expression of target HDAC genes or a target HDAC gene family, wherein the HDAC genes or HDAC gene family sequences share sequence homology. Such homologous sequences can be identified as is known in the art, for example using sequence alignments. siNA molecules can be designed to target such homologous HDAC sequences, for example using perfectly complementary sequences or by incorporating non-canonical base pairs, for example mismatches and/or wobble base pairs, that can provide additional target sequences. In instances where mismatches are identified, non-canonical base pairs (for example, mismatches and/or wobble bases) can be used to generate siNA molecules that target more than one gene sequence. In a non-limiting example, non-canonical base pairs such as UU and CC base pairs are used to generate siNA molecules that are capable of targeting sequences for differing polynucleotide targets that share sequence homology. As such, one advantage of using siNAs of the invention is that a single siNA can be designed to include nucleic acid sequence that is complementary to the nucleotide sequence that is conserved between the homologous genes. In this approach, a single siNA can be used to inhibit expression of more than one gene instead of using more than one siNA molecule to target the different genes.

In one embodiment, the invention features siNA molecules that target conserved HDAC nucleotide sequences. The conserved HDAC sequences can be conserved across class I HDAC targets (e.g., any of HDAC 1, 2, 3 and/or 8), class II HDAC targets (e.g., any of HDAC 4, 5, 6, 7, 9a, 9b, and/or 10), class III targets (SIR T1, 2, 3, 4, 5, 6, and/or 7), or any combination thereof (e.g., any of HDAC 1, 2, 3, 4, 5, 6, 7, 8, 9a, 9b, 10, and/or 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7).

In one embodiment, the invention features a siNA molecule having RNAi activity against target HDAC RNA (e.g., coding or non-coding RNA), wherein the siNA molecule comprises a sequence complementary to any HDAC RNA sequence, such as those sequences having HDAC GenBank Accession Nos. shown in Table I, or in U.S. Ser. No. 10/923,536 and U.S. Ser. No. 10/923,536, both incorporated by reference herein. In another embodiment, the invention features a siNA molecule having RNAi activity against target HDAC RNA, wherein the siNA molecule comprises a sequence complementary to an RNA having HDAC variant encoding sequence, for example other mutant HDAC genes known in the art to be associated with the maintenance and/or development of diseases, traits, disorders, and/or conditions described herein (e.g., cancer and proliferative diseases) or otherwise known in the art. Chemical modifications as shown in Table IV or otherwise described herein can be applied to any siNA construct of the invention. In another embodiment, a siNA molecule of the invention includes a nucleotide sequence that can interact with nucleotide sequence of a target HDAC gene and thereby mediate silencing of target HDAC gene expression, for example, wherein the siNA mediates regulation of target HDAC gene expression by cellular processes that modulate the chromatin structure or methylation patterns of the target gene and prevent transcription of the target HDAC gene.

In one embodiment, siNA molecules of the invention are used to down regulate or inhibit the expression of HDAC proteins arising from haplotype polymorphisms that are associated with a trait, disease or condition in a subject or organism, such as cancer or proliferative diseases and conditions. Analysis of HDAC genes, or HDAC protein or RNA levels can be used to identify subjects with such polymorphisms or those subjects who are at risk of developing traits, conditions, or diseases described herein. These subjects are amenable to treatment, for example, treatment with siNA molecules of the invention and any other composition useful in treating diseases related to target gene expression. As such, analysis of HDAC protein or RNA levels can be used to determine treatment type and the course of therapy in treating a subject. Monitoring of HDAC protein or RNA levels can be used to predict treatment outcome and to determine the efficacy of compounds and compositions that modulate the level and/or activity of certain HDAC proteins associated with a trait, disorder, condition, or disease (e.g., cancer and/or proliferative diseases and conditions).

In one embodiment of the invention a siNA molecule comprises an antisense strand comprising a nucleotide sequence that is complementary to a target HDAC nucleotide sequence or a portion thereof. The siNA further comprises a sense strand, wherein said sense strand comprises a nucleotide sequence of a target HDAC gene or a portion thereof.

In another embodiment, a siNA molecule comprises an antisense region comprising a nucleotide sequence that is complementary to a nucleotide sequence encoding a target HDAC protein or a portion thereof. The siNA molecule further comprises a sense region, wherein said sense region comprises a nucleotide sequence of a target HDAC gene or a portion thereof.

In another embodiment, the invention features a siNA molecule comprising nucleotide sequence, for example, nucleotide sequence in the antisense region of the siNA molecule that is complementary to a nucleotide sequence or portion of sequence of a target HDAC gene. In another embodiment, the invention features a siNA molecule comprising a region, for example, the antisense region of the siNA construct, complementary to a sequence comprising a target HDAC gene sequence or a portion thereof.

In yet another embodiment, the invention features a siNA molecule comprising a sequence, for example, the antisense sequence of the siNA construct, complementary to a sequence or portion of sequence comprising sequence represented by GenBank Accession Nos. shown in Table I or in U.S. Ser. No. 10/923,536 and U.S. Ser. No. 10/923,536, both incorporated by reference herein. Chemical modifications in Table IV and otherwise described herein can be applied to any siNA construct of the invention.

In one embodiment of the invention a siNA molecule comprises an antisense strand having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, wherein the antisense strand is complementary to a target HDAC RNA sequence or a portion thereof, and wherein said siNA further comprises a sense strand having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, and wherein said sense strand and said antisense strand are distinct nucleotide sequences where at least about 15 nucleotides in each strand are complementary to the other strand.

In another embodiment of the invention a siNA molecule of the invention comprises an antisense region having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, wherein the antisense region is complementary to a target HDAC DNA sequence, and wherein said siNA further comprises a sense region having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, wherein said sense region and said antisense region are comprised in a linear molecule where the sense region comprises at least about 15 nucleotides that are complementary to the antisense region.

In one embodiment, a siNA molecule of the invention has RNAi activity that modulates expression of HDAC RNA encoded by one or more HDAC genes. Because various HDAC genes can share some degree of sequence homology with each other, siNA molecules can be designed to target a class of HDAC genes (e.g., class I, class II, and/or class III HDAC genes) or alternately specific genes (e.g., any of HDAC 1, 2, 3, 4, 5, 6, 7, 8, 9a, 9b, 10, and/or 11, and/or SIR T1, 2, 3, 4, 5, 6 and/or 7 or polymorphic variants thereof) by selecting sequences that are either shared amongst different HDAC gene targets or alternatively that are unique for a specific HDAC gene target. Therefore, in one embodiment, the siNA molecule can be designed to target conserved regions of target HDAC RNA sequences having homology among several gene variants so as to target a class of HDAC genes with one siNA molecule. Accordingly, in one embodiment, the siNA molecule of the invention modulates the expression of one or both HDAC gene alleles in a subject. In another embodiment, the siNA molecule can be designed to target a sequence that is unique to a specific target HDAC RNA sequence (e.g., a single allele or single nucleotide polymorphism (SNP)) due to the high degree of specificity that the siNA molecule requires to mediate RNAi activity.

In one embodiment, nucleic acid molecules of the invention that act as mediators of the RNA interference gene silencing response are double-stranded nucleic acid molecules. In another embodiment, the siNA molecules of the invention consist of duplex nucleic acid molecules containing about 15 to about 30 base pairs between oligonucleotides comprising about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides. In yet another embodiment, siNA molecules of the invention comprise duplex nucleic acid molecules with overhanging ends of about 1 to about 3 (e.g., about 1, 2, or 3) nucleotides, for example, about 21-nucleotide duplexes with about 19 base pairs and 3′-terminal mononucleotide, dinucleotide, or trinucleotide overhangs. In yet another embodiment, siNA molecules of the invention comprise duplex nucleic acid molecules with blunt ends, where both ends are blunt, or alternatively, where one of the ends is blunt.

In one embodiment, the invention features one or more chemically-modified siNA constructs having specificity for target HDAC nucleic acid molecules, such as HDAC DNA, or HDAC RNA encoding a HDAC protein or non-coding RNA associated with the expression of target HDAC genes.

In one embodiment, the invention features a RNA based siNA molecule (e.g., a siNA comprising 2′-OH nucleotides) having specificity for nucleic acid molecules that includes one or more chemical modifications described herein. Non-limiting examples of such chemical modifications include without limitation phosphorothioate internucleotide linkages, 2′-deoxyribonucleotides, 2′-O-methyl ribonucleotides, 2′-deoxy-2′-fluoro ribonucleotides, 4′-thio ribonucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides (see for example U.S. Ser. No. 10/981,966 filed Nov. 5, 2004, incorporated by reference herein), “universal base” nucleotides, “acyclic” nucleotides, 5-C-methyl nucleotides, and terminal glyceryl and/or inverted deoxy abasic residue incorporation. These chemical modifications, when used in various siNA constructs, (e.g., RNA based siNA constructs), are shown to preserve RNAi activity in cells while at the same time, dramatically increasing the serum stability of these compounds. Furthermore, contrary to the data published by Parrish et al., supra, applicant demonstrates that multiple (greater than one) phosphorothioate substitutions are well-tolerated and confer substantial increases in serum stability for modified siNA constructs.

In one embodiment, a siNA molecule of the invention comprises modified nucleotides while maintaining the ability to mediate RNAi. The modified nucleotides can be used to improve in vitro or in vivo characteristics such as stability, activity, toxicity, immune response, and/or bioavailability. For example, a siNA molecule of the invention can comprise modified nucleotides as a percentage of the total number of nucleotides present in the siNA molecule. As such, a siNA molecule of the invention can generally comprise about 5% to about 100% modified nucleotides (e.g., about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% modified nucleotides). For example, in one embodiment, between about 5% to about 100% (e.g., about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% modified nucleotides) of the nucleotide positions in a siNA molecule of the invention comprise a nucleic acid sugar modification, such as a 2′-sugar modification, e.g., 2′-O-methyl nucleotides, 2′-deoxy-2′-fluoro nucleotides, 2′-O-methoxyethyl nucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides, or 2′-deoxy nucleotides. In another embodiment, between about 5% to about 100% (e.g., about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% modified nucleotides) of the nucleotide positions in a siNA molecule of the invention comprise a nucleic acid base modification, such as inosine, purine, pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2,4,6-trimethoxy benzene, 3-methyl uracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidines (e.g., 5-methylcytidine), 5-alkyluridines (e.g., ribothymidine), 5-halouridine (e.g., 5-bromouridine) or 6-azapyrimidines or 6-alkylpyrimidines (e.g. 6-methyluridine), or propyne modifications. In another embodiment, between about 5% to about 100% (e.g., about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% modified nucleotides) of the nucleotide positions in a siNA molecule of the invention comprise a nucleic acid backbone modification, such as a backbone modification having Formula I herein. In another embodiment, between about 5% to about 100% (e.g., about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% modified nucleotides) of the nucleotide positions in a siNA molecule of the invention comprise a nucleic acid sugar, base, or backbone modification or any combination thereof (e.g., any combination of nucleic acid sugar, base, backbone or non-nucleotide modifications herein). The actual percentage of modified nucleotides present in a given siNA molecule will depend on the total number of nucleotides present in the siNA. If the siNA molecule is single stranded, the percent modification can be based upon the total number of nucleotides present in the single stranded siNA molecules. Likewise, if the siNA molecule is double stranded, the percent modification can be based upon the total number of nucleotides present in the sense strand, antisense strand, or both the sense and antisense strands.

A siNA molecule of the invention can comprise modified nucleotides at various locations within the siNA molecule. In one embodiment, a double stranded siNA molecule of the invention comprises modified nucleotides at internal base paired positions within the siNA duplex. For example, internal positions can comprise positions from about 3 to about 19 nucleotides from the 5′-end of either sense or antisense strand or region of a 21 nucleotide siNA duplex having 19 base pairs and two nucleotide 3′-overhangs. In another embodiment, a double stranded siNA molecule of the invention comprises modified nucleotides at non-base paired or overhang regions of the siNA molecule. For example, overhang positions can comprise positions from about 20 to about 21 nucleotides from the 5′-end of either sense or antisense strand or region of a 21 nucleotide siNA duplex having 19 base pairs and two nucleotide 3′-overhangs. In another embodiment, a double stranded siNA molecule of the invention comprises modified nucleotides at terminal positions of the siNA molecule. For example, such terminal regions include the 3′-position, 5′-position, for both 3′ and 5′-positions of the sense and/or antisense strand or region of the siNA molecule. In another embodiment, a double stranded siNA molecule of the invention comprises modified nucleotides at base-paired or internal positions, non-base paired or overhang regions, and/or terminal regions, or any combination thereof.

One aspect of the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a target HDAC gene or that directs cleavage of a target HDAC RNA. In one embodiment, the double stranded siNA molecule comprises one or more chemical modifications and each strand of the double-stranded siNA is about 21 nucleotides long. In one embodiment, the double-stranded siNA molecule does not contain any ribonucleotides. In another embodiment, the double-stranded siNA molecule comprises one or more ribonucleotides. In one embodiment, each strand of the double-stranded siNA molecule independently comprises about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, wherein each strand comprises about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides that are complementary to the nucleotides of the other strand. In one embodiment, one of the strands of the double-stranded siNA molecule comprises a nucleotide sequence that is complementary to a nucleotide sequence or a portion thereof of the target HDAC gene, and the second strand of the double-stranded siNA molecule comprises a nucleotide sequence substantially similar to the nucleotide sequence of the target HDAC gene or a portion thereof.

In another embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a target HDAC gene or that directs cleavage of a target HDAC RNA, comprising an antisense region, wherein the antisense region comprises a nucleotide sequence that is complementary to a nucleotide sequence of the target gene or a portion thereof, and a sense region, wherein the sense region comprises a nucleotide sequence substantially similar to the nucleotide sequence of the target HDAC gene or a portion thereof. In one embodiment, the antisense region and the sense region independently comprise about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, wherein the antisense region comprises about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides that are complementary to nucleotides of the sense region.

In one embodiment, a siNA molecule of the invention comprises blunt ends, i.e., ends that do not include any overhanging nucleotides. For example, a siNA molecule comprising modifications described herein (e.g., comprising nucleotides having Formulae I-VII or siNA constructs comprising “Stab 00“−”Stab 34” or “Stab 3F“−”Stab 34F” (Table IV) or any combination thereof (see Table IV)) and/or any length described herein can comprise blunt ends or ends with no overhanging nucleotides.

In one embodiment, any siNA molecule of the invention can comprise one or more blunt ends, i.e. where a blunt end does not have any overhanging nucleotides. In one embodiment, the blunt ended siNA molecule has a number of base pairs equal to the number of nucleotides present in each strand of the siNA molecule. In another embodiment, the siNA molecule comprises one blunt end, for example wherein the 5′-end of the antisense strand and the 3′-end of the sense strand do not have any overhanging nucleotides. In another example, the siNA molecule comprises one blunt end, for example wherein the 3′-end of the antisense strand and the 5′-end of the sense strand do not have any overhanging nucleotides. In another example, a siNA molecule comprises two blunt ends, for example wherein the 3′-end of the antisense strand and the 5′-end of the sense strand as well as the 5′-end of the antisense strand and 3′-end of the sense strand do not have any overhanging nucleotides. A blunt ended siNA molecule can comprise, for example, from about 15 to about 30 nucleotides (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides). Other nucleotides present in a blunt ended siNA molecule can comprise, for example, mismatches, bulges, loops, or wobble base pairs to modulate the activity of the siNA molecule to mediate RNA interference.

By “blunt ends” is meant symmetric termini or termini of a double stranded siNA molecule having no overhanging nucleotides. The two strands of a double stranded siNA molecule align with each other without over-hanging nucleotides at the termini. For example, a blunt ended siNA construct comprises terminal nucleotides that are complementary between the sense and antisense regions of the siNA molecule.

In one embodiment, a siNA molecule of the invention is a double-stranded short interfering nucleic acid (siNA), wherein the double stranded nucleic acid molecule comprises about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) base pairs, and wherein one or more (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) of the nucleotide positions in each strand of the siNA molecule comprises a chemical modification. In another embodiment, the siNA contains at least 2, 3, 4, 5, or more different chemical modifications.

In one embodiment, the invention features double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a target HDAC gene or that directs cleavage of a target HDAC RNA, wherein the siNA molecule comprises about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) base pairs, and wherein each strand of the siNA molecule comprises one or more chemical modifications. In one embodiment, each strand of the double stranded siNA molecule comprises at least two (e.g., 2, 3, 4, 5, or more) different chemical modifications, e.g., different nucleotide sugar, base, or backbone modifications. In another embodiment, one of the strands of the double-stranded siNA molecule comprises a nucleotide sequence that is complementary to a nucleotide sequence of a target HDAC gene or a portion thereof, and the second strand of the double-stranded siNA molecule comprises a nucleotide sequence substantially similar to the nucleotide sequence or a portion thereof of the target HDAC gene. In another embodiment, one of the strands of the double-stranded siNA molecule comprises a nucleotide sequence that is complementary to a nucleotide sequence of a target HDAC gene or portion thereof, and the second strand of the double-stranded siNA molecule comprises a nucleotide sequence substantially similar to the nucleotide sequence or portion thereof of the target HDAC gene. In another embodiment, each strand of the siNA molecule comprises about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, and each strand comprises at least about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides that are complementary to the nucleotides of the other strand. The target HDAC gene can comprise, for example, sequences referred to herein or incorporated herein by reference.

In one embodiment, each strand of a double stranded siNA molecule of the invention comprises a different pattern of chemical modifications, such as any “Stab 00”−“Stab 34” or “Stab 3F“−”Stab 34F” (Table IV) modification patterns herein or any combination thereof (see Table IV). Non-limiting examples of sense and antisense strands of such siNA molecules having various modification patterns are shown in Table III.

In one embodiment, a siNA molecule of the invention comprises no ribonucleotides. In another embodiment, a siNA molecule of the invention comprises ribonucleotides.

In one embodiment, a siNA molecule of the invention comprises an antisense region comprising a nucleotide sequence that is complementary to a nucleotide sequence of a target HDAC gene or a portion thereof, and the siNA further comprises a sense region comprising a nucleotide sequence substantially similar to the nucleotide sequence of the target HDAC gene or a portion thereof. In another embodiment, the antisense region and the sense region each comprise about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides and the antisense region comprises at least about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides that are complementary to nucleotides of the sense region. In one embodiment, each strand of the double stranded siNA molecule comprises at least two (e.g., 2, 3, 4, 5, or more) different chemical modifications, e.g., different nucleotide sugar, base, or backbone modifications. The target HDAC gene can comprise, for example, sequences referred to herein or incorporated by reference herein. In another embodiment, the siNA is a double stranded nucleic acid molecule, where each of the two strands of the siNA molecule independently comprise about 15 to about 40 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 23, 33, 34, 35, 36, 37, 38, 39, or 40) nucleotides, and where one of the strands of the siNA molecule comprises at least about 15 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 or more) nucleotides that are complementary to the nucleic acid sequence of the target gene or a portion thereof.

In one embodiment, a siNA molecule of the invention comprises a sense region and an antisense region, wherein the antisense region comprises a nucleotide sequence that is complementary to a nucleotide sequence of RNA encoded by a target HDAC gene, or a portion thereof, and the sense region comprises a nucleotide sequence that is complementary to the antisense region. In one embodiment, the siNA molecule is assembled from two separate oligonucleotide fragments, wherein one fragment comprises the sense region and the second fragment comprises the antisense region of the siNA molecule. In another embodiment, the sense region is connected to the antisense region via a linker molecule. In another embodiment, the sense region is connected to the antisense region via a linker molecule, such as a nucleotide or non-nucleotide linker. In one embodiment, each strand of the double stranded siNA molecule comprises at least two (e.g., 2, 3, 4, 5, or more) different chemical modifications, e.g., different nucleotide sugar, base, or backbone modifications. The target HDAC gene can comprise, for example, sequences referred to herein, incorporated by reference herein, or otherwise known in the art.

In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a target HDAC gene or that directs cleavage of a target HDAC RNA, comprising a sense region and an antisense region, wherein the antisense region comprises a nucleotide sequence that is complementary to a nucleotide sequence of RNA encoded by the target HDAC gene or a portion thereof and the sense region comprises a nucleotide sequence that is complementary to the antisense region, and wherein the siNA molecule has one or more modified pyrimidine and/or purine nucleotides. In one embodiment, each strand of the double stranded siNA molecule comprises at least two (e.g., 2, 3, 4, 5, or more) different chemical modifications, e.g., different nucleotide sugar, base, or backbone modifications. In one embodiment, the pyrimidine nucleotides in the sense region are 2′-O-methyl pyrimidine nucleotides or 2′-deoxy-2′-fluoro pyrimidine nucleotides and the purine nucleotides present in the sense region are 2′-deoxy purine nucleotides. In another embodiment, the pyrimidine nucleotides in the sense region are 2′-deoxy-2′-fluoro pyrimidine nucleotides and the purine nucleotides present in the sense region are 2′-O-methyl purine nucleotides. In another embodiment, the pyrimidine nucleotides in the sense region are 2′-deoxy-2′-fluoro pyrimidine nucleotides and the purine nucleotides present in the sense region are 2′-deoxy purine nucleotides. In one embodiment, the pyrimidine nucleotides in the antisense region are 2′-deoxy-2′-fluoro pyrimidine nucleotides and the purine nucleotides present in the antisense region are 2′-O-methyl or 2′-deoxy purine nucleotides. In another embodiment of any of the above-described siNA molecules, any nucleotides present in a non-complementary region of the sense strand (e.g. overhang region) are 2′-deoxy nucleotides.

In one embodiment, the invention features a siNA molecule comprising at least one modified nucleotide, wherein the modified nucleotide is a 2′-deoxy-2′-fluoro nucleotide, 2′-O-trifluoromethyl nucleotide, 2′-O-ethyl-trifluoromethoxy nucleotide, or 2′-O-difluoromethoxy-ethoxy nucleotide or any other modified nucleoside/nucleotide described herein and in U.S. Ser. No. 10/981,966, filed Nov. 5, 2004, incorporated by reference herein. In one embodiment, the invention features a siNA molecule comprising at least two (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) modified nucleotides, wherein the modified nucleotide is selected from the group consisting of 2′-deoxy-2′-fluoro nucleotide, 2′-O-trifluoromethyl nucleotide, 2′-O-ethyl-trifluoromethoxy nucleotide, or 2′-O-difluoromethoxy-ethoxy nucleotide or any other modified nucleoside/nucleotide described herein and in U.S. Ser. No. 10/981,966, filed Nov. 5, 2004, incorporated by reference herein. The modified nucleotide/nucleoside can be the same or different. The siNA can be, for example, about 15 to about 40 nucleotides in length. In one embodiment, all pyrimidine nucleotides present in the siNA are 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy, 4′-thio pyrimidine nucleotides. In one embodiment, the modified nucleotides in the siNA include at least one 2′-deoxy-2′-fluoro cytidine or 2′-deoxy-2′-fluoro uridine nucleotide. In another embodiment, the modified nucleotides in the siNA include at least one 2′-fluoro cytidine and at least one 2′-deoxy-2′-fluoro uridine nucleotides. In one embodiment, all uridine nucleotides present in the siNA are 2′-deoxy-2′-fluoro uridine nucleotides. In one embodiment, all cytidine nucleotides present in the siNA are 2′-deoxy-2′-fluoro cytidine nucleotides. In one embodiment, all adenosine nucleotides present in the siNA are 2′-deoxy-2′-fluoro adenosine nucleotides. In one embodiment, all guanosine nucleotides present in the siNA are 2′-deoxy-2′-fluoro guanosine nucleotides. The siNA can further comprise at least one modified internucleotidic linkage, such as phosphorothioate linkage. In one embodiment, the 2′-deoxy-2′-fluoronucleotides are present at specifically selected locations in the siNA that are sensitive to cleavage by ribonucleases, such as locations having pyrimidine nucleotides.

In one embodiment, the invention features a method of increasing the stability of a siNA molecule against cleavage by ribonucleases comprising introducing at least one modified nucleotide into the siNA molecule, wherein the modified nucleotide is a 2′-deoxy-2′-fluoro nucleotide. In one embodiment, all pyrimidine nucleotides present in the siNA are 2′-deoxy-2′-fluoro pyrimidine nucleotides. In one embodiment, the modified nucleotides in the siNA include at least one 2′-deoxy-2′-fluoro cytidine or 2′-deoxy-2′-fluoro uridine nucleotide. In another embodiment, the modified nucleotides in the siNA include at least one 2′-fluoro cytidine and at least one 2′-deoxy-2′-fluoro uridine nucleotides. In one embodiment, all uridine nucleotides present in the siNA are 2′-deoxy-2′-fluoro uridine nucleotides. In one embodiment, all cytidine nucleotides present in the siNA are 2′-deoxy-2′-fluoro cytidine nucleotides. In one embodiment, all adenosine nucleotides present in the siNA are 2′-deoxy-2′-fluoro adenosine nucleotides. In one embodiment, all guanosine nucleotides present in the siNA are 2′-deoxy-2′-fluoro guanosine nucleotides. The siNA can further comprise at least one modified internucleotidic linkage, such as a phosphorothioate linkage. In one embodiment, the 2′-deoxy-2′-fluoronucleotides are present at specifically selected locations in the siNA that are sensitive to cleavage by ribonucleases, such as locations having pyrimidine nucleotides.

In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a target HDAC gene or that directs cleavage of a target HDAC RNA, comprising a sense region and an antisense region, wherein the antisense region comprises a nucleotide sequence that is complementary to a nucleotide sequence of RNA encoded by the target HDAC gene or a portion thereof and the sense region comprises a nucleotide sequence that is complementary to the antisense region, and wherein the purine nucleotides present in the antisense region comprise 2′-deoxy-purine nucleotides. In an alternative embodiment, the purine nucleotides present in the antisense region comprise 2′-O-methyl purine nucleotides. In either of the above embodiments, the antisense region can comprise a phosphorothioate internucleotide linkage at the 3′ end of the antisense region. Alternatively, in either of the above embodiments, the antisense region can comprise a glyceryl modification at the 3′ end of the antisense region. In another embodiment of any of the above-described siNA molecules, any nucleotides present in a non-complementary region of the antisense strand (e.g. overhang region) are 2′-deoxy nucleotides.

In one embodiment, the antisense region of a siNA molecule of the invention comprises sequence complementary to a portion of an endogenous transcript having sequence unique to a particular disease or trait related allele in a subject or organism, such as sequence comprising a single nucleotide polymorphism (SNP) associated with the disease or trait specific allele. As such, the antisense region of a siNA molecule of the invention can comprise sequence complementary to sequences that are unique to a particular allele to provide specificity in mediating selective RNAi against the disease, condition, or trait related allele.

In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a target HDAC gene or that directs cleavage of a target HDAC RNA, wherein the siNA molecule is assembled from two separate oligonucleotide fragments wherein one fragment comprises the sense region and the second fragment comprises the antisense region of the siNA molecule. In one embodiment, each strand of the double stranded siNA molecule is about 21 nucleotides long and where about 19 nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule, wherein at least two 3′ terminal nucleotides of each fragment of the siNA molecule are not base-paired to the nucleotides of the other fragment of the siNA molecule. In another embodiment, the siNA molecule is a double stranded nucleic acid molecule, where each strand is about 19 nucleotide long and where the nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule to form at least about 15 (e.g., 15, 16, 17, 18, or 19) base pairs, wherein one or both ends of the siNA molecule are blunt ends. In one embodiment, each of the two 3′ terminal nucleotides of each fragment of the siNA molecule is a 2′-deoxy-pyrimidine nucleotide, such as a 2′-deoxy-thymidine. In another embodiment, all nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule. In another embodiment, the siNA molecule is a double stranded nucleic acid molecule of about 19 to about 25 base pairs having a sense region and an antisense region, where about 19 nucleotides of the antisense region are base-paired to the nucleotide sequence or a portion thereof of the RNA encoded by the target gene. In another embodiment, about 21 nucleotides of the antisense region are base-paired to the nucleotide sequence or a portion thereof of the RNA encoded by the target gene. In any of the above embodiments, the 5′-end of the fragment comprising said antisense region can optionally include a phosphate group.

In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that inhibits the expression of a target HDAC RNA sequence, wherein the siNA molecule does not contain any ribonucleotides and wherein each strand of the double-stranded siNA molecule is about 15 to about 30 nucleotides. In one embodiment, the siNA molecule is 21 nucleotides in length. Examples of non-ribonucleotide containing siNA constructs are combinations of stabilization chemistries shown in Table IV in any combination of Sense/Antisense chemistries, such as Stab 7/8, Stab 7/11, Stab 8/8, Stab 18/8, Stab 18/11, Stab 12/13, Stab 7/13, Stab 18/13, Stab 7/19, Stab 8/19, Stab 18/19, Stab 7/20, Stab 8/20, Stab 18/20, Stab 7/32, Stab 8/32, or Stab 18/32 (e.g., any siNA having Stab 7, 8, 11, 12, 13, 14, 15, 17, 18, 19, 20, or 32 sense or antisense strands or any combination thereof). Herein, numeric Stab chemistries can include both 2′-fluoro and 2′-OCF3 versions of the chemistries shown in Table IV. For example, “Stab 7/8” refers to both Stab 7/8 and Stab 7F/8F etc. In one embodiment, the invention features a chemically synthesized double stranded RNA molecule that directs cleavage of a target HDAC RNA via RNA interference, wherein each strand of said RNA molecule is about 15 to about 30 nucleotides in length; one strand of the RNA molecule comprises nucleotide sequence having sufficient complementarity to the target HDAC RNA for the RNA molecule to direct cleavage of the target HDAC RNA via RNA interference; and wherein at least one strand of the RNA molecule optionally comprises one or more chemically modified nucleotides described herein, such as without limitation deoxynucleotides, 2′-O-methyl nucleotides, 2′-deoxy-2′-fluoro nucleotides, 2′-O-methoxyethyl nucleotides, 4′-thio nucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides, etc. The chemically modified nucleotides can be the same or different.

In one embodiment, a target HDAC RNA of the invention comprises sequence encoding a HDAC protein.

In one embodiment, target HDAC RNA of the invention comprises non-coding HDAC RNA sequence (e.g., miRNA, snRNA siRNA etc.).

In one embodiment, the invention features a medicament comprising a siNA molecule of the invention.

In one embodiment, the invention features an active ingredient comprising a siNA molecule of the invention.

In one embodiment, the invention features the use of a double-stranded short interfering nucleic acid (siNA) molecule to inhibit, down-regulate, or reduce expression of a target HDAC gene, wherein the siNA molecule comprises one or more chemical modifications that can be the same or different and each strand of the double-stranded siNA is independently about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 or more) nucleotides long. In one embodiment, the siNA molecule of the invention is a double stranded nucleic acid molecule comprising one or more chemical modifications, where each of the two fragments of the siNA molecule independently comprise about 15 to about 40 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 23, 33, 34, 35, 36, 37, 38, 39, or 40) nucleotides and where one of the strands comprises at least 15 nucleotides that are complementary to nucleotide sequence of HDAC target encoding RNA or a portion thereof. In a non-limiting example, each of the two fragments of the siNA molecule comprise about 21 nucleotides. In another embodiment, the siNA molecule is a double stranded nucleic acid molecule comprising one or more chemical modifications, where each strand is about 21 nucleotide long and where about 19 nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule, wherein at least two 3′ terminal nucleotides of each fragment of the siNA molecule are not base-paired to the nucleotides of the other fragment of the siNA molecule. In another embodiment, the siNA molecule is a double stranded nucleic acid molecule comprising one or more chemical modifications, where each strand is about 19 nucleotide long and where the nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule to form at least about 15 (e.g., 15, 16, 17, 18, or 19) base pairs, wherein one or both ends of the siNA molecule are blunt ends. In one embodiment, each of the two 3′ terminal nucleotides of each fragment of the siNA molecule is a 2′-deoxy-pyrimidine nucleotide, such as a 2′-deoxy-thymidine. In another embodiment, all nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule. In another embodiment, the siNA molecule is a double stranded nucleic acid molecule of about 19 to about 25 base pairs having a sense region and an antisense region and comprising one or more chemical modifications, where about 19 nucleotides of the antisense region are base-paired to the nucleotide sequence or a portion thereof of the RNA encoded by the target HDAC gene. In another embodiment, about 21 nucleotides of the antisense region are base-paired to the nucleotide sequence or a portion thereof of the RNA encoded by the target HDAC gene. In any of the above embodiments, the 5′-end of the fragment comprising said antisense region can optionally include a phosphate group.

In one embodiment, the invention features the use of a double-stranded short interfering nucleic acid (siNA) molecule that inhibits, down-regulates, or reduces expression of a target HDAC gene, wherein one of the strands of the double-stranded siNA molecule is an antisense strand which comprises nucleotide sequence that is complementary to nucleotide sequence of target HDAC RNA or a portion thereof, the other strand is a sense strand which comprises nucleotide sequence that is complementary to a nucleotide sequence of the antisense strand. In one embodiment, each strand has at least two (e.g., 2, 3, 4, 5, or more) chemical modifications, which can be the same or different, such as nucleotide, sugar, base, or backbone modifications. In one embodiment, a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification. In one embodiment, a majority of the purine nucleotides present in the double-stranded siNA molecule comprises a sugar modification.

In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that inhibits, down-regulates, or reduces expression of a target HDAC gene, wherein one of the strands of the double-stranded siNA molecule is an antisense strand which comprises nucleotide sequence that is complementary to nucleotide sequence of target HDAC RNA that encodes a protein or portion thereof, the other strand is a sense strand which comprises nucleotide sequence that is complementary to a nucleotide sequence of the antisense strand and wherein a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification. In one embodiment, each strand of the siNA molecule comprises about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more) nucleotides, wherein each strand comprises at least about 15 nucleotides that are complementary to the nucleotides of the other strand. In one embodiment, the siNA molecule is assembled from two oligonucleotide fragments, wherein one fragment comprises the nucleotide sequence of the antisense strand of the siNA molecule and a second fragment comprises nucleotide sequence of the sense region of the siNA molecule. In one embodiment, the sense strand is connected to the antisense strand via a linker molecule, such as a polynucleotide linker or a non-nucleotide linker. In a further embodiment, the pyrimidine nucleotides present in the sense strand are 2′-deoxy-2′fluoro pyrimidine nucleotides and the purine nucleotides present in the sense region are 2′-deoxy purine nucleotides. In another embodiment, the pyrimidine nucleotides present in the sense strand are 2′-deoxy-2′fluoro pyrimidine nucleotides and the purine nucleotides present in the sense region are 2′-O-methyl purine nucleotides. In still another embodiment, the pyrimidine nucleotides present in the antisense strand are 2′-deoxy-2′-fluoro pyrimidine nucleotides and any purine nucleotides present in the antisense strand are 2′-deoxy purine nucleotides. In another embodiment, the antisense strand comprises one or more 2′-deoxy-2′-fluoro pyrimidine nucleotides and one or more 2′-O-methyl purine nucleotides. In another embodiment, the pyrimidine nucleotides present in the antisense strand are 2′-deoxy-2′-fluoro pyrimidine nucleotides and any purine nucleotides present in the antisense strand are 2′-O-methyl purine nucleotides. In a further embodiment the sense strand comprises a 3′-end and a 5′-end, wherein a terminal cap moiety (e.g., an inverted deoxy abasic moiety or inverted deoxy nucleotide moiety such as inverted thymidine) is present at the 5′-end, the 3′-end, or both of the 5′ and 3′ ends of the sense strand. In another embodiment, the antisense strand comprises a phosphorothioate internucleotide linkage at the 3′ end of the antisense strand. In another embodiment, the antisense strand comprises a glyceryl modification at the 3′ end. In another embodiment, the 5′-end of the antisense strand optionally includes a phosphate group.

In any of the above-described embodiments of a double-stranded short interfering nucleic acid (siNA) molecule that inhibits expression of a target HDAC gene, wherein a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification, each of the two strands of the siNA molecule can comprise about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more) nucleotides. In one embodiment, about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more) nucleotides of each strand of the siNA molecule are base-paired to the complementary nucleotides of the other strand of the siNA molecule. In another embodiment, about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more) nucleotides of each strand of the siNA molecule are base-paired to the complementary nucleotides of the other strand of the siNA molecule, wherein at least two 3′ terminal nucleotides of each strand of the siNA molecule are not base-paired to the nucleotides of the other strand of the siNA molecule. In another embodiment, each of the two 3′ terminal nucleotides of each fragment of the siNA molecule is a 2′-deoxy-pyrimidine, such as 2′-deoxy-thymidine. In one embodiment, each strand of the siNA molecule is base-paired to the complementary nucleotides of the other strand of the siNA molecule. In one embodiment, about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides of the antisense strand are base-paired to the nucleotide sequence of the target RNA or a portion thereof. In one embodiment, about 18 to about 25 (e.g., about 18, 19, 20, 21, 22, 23, 24, or 25) nucleotides of the antisense strand are base-paired to the nucleotide sequence of the target RNA or a portion thereof.

In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that inhibits expression of a target HDAC gene, wherein one of the strands of the double-stranded siNA molecule is an antisense strand which comprises nucleotide sequence that is complementary to nucleotide sequence of target HDAC RNA or a portion thereof, the other strand is a sense strand which comprises nucleotide sequence that is complementary to a nucleotide sequence of the antisense strand. In one embodiment, each strand has at least two (e.g., 2, 3, 4, 5, or more) different chemical modifications, such as nucleotide sugar, base, or backbone modifications. In one embodiment, a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification. In one embodiment, a majority of the purine nucleotides present in the double-stranded siNA molecule comprises a sugar modification. In one embodiment, the 5′-end of the antisense strand optionally includes a phosphate group.

In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that inhibits expression of a target HDAC gene, wherein one of the strands of the double-stranded siNA molecule is an antisense strand which comprises nucleotide sequence that is complementary to nucleotide sequence of target HDAC RNA or a portion thereof, the other strand is a sense strand which comprises nucleotide sequence that is complementary to a nucleotide sequence of the antisense strand and wherein a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification, and wherein the nucleotide sequence or a portion thereof of the antisense strand is complementary to a nucleotide sequence of the untranslated region or a portion thereof of the target RNA.

In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that inhibits expression of a target HDAC gene, wherein one of the strands of the double-stranded siNA molecule is an antisense strand which comprises nucleotide sequence that is complementary to nucleotide sequence of target HDAC RNA or a portion thereof, wherein the other strand is a sense strand which comprises nucleotide sequence that is complementary to a nucleotide sequence of the antisense strand, wherein a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification, and wherein the nucleotide sequence of the antisense strand is complementary to a nucleotide sequence of the target HDAC RNA or a portion thereof that is present in the target HDAC RNA.

In one embodiment, the invention features a composition comprising a siNA molecule of the invention in a pharmaceutically acceptable carrier or diluent.

In a non-limiting example, the introduction of chemically-modified nucleotides into nucleic acid molecules provides a powerful tool in overcoming potential limitations of in vivo stability and bioavailability inherent to native RNA molecules that are delivered exogenously. For example, the use of chemically-modified nucleic acid molecules can enable a lower dose of a particular nucleic acid molecule for a given therapeutic effect since chemically-modified nucleic acid molecules tend to have a longer half-life in serum. Furthermore, certain chemical modifications can improve the bioavailability of nucleic acid molecules by targeting particular cells or tissues and/or improving cellular uptake of the nucleic acid molecule. Therefore, even if the activity of a chemically-modified nucleic acid molecule is reduced as compared to a native nucleic acid molecule, for example, when compared to an all-RNA nucleic acid molecule, the overall activity of the modified nucleic acid molecule can be greater than that of the native molecule due to improved stability and/or delivery of the molecule. Unlike native unmodified siNA, chemically-modified siNA can also minimize the possibility of activating interferon activity or immunostimulation in humans.

In any of the embodiments of siNA molecules described herein, the antisense region of a siNA molecule of the invention can comprise a phosphorothioate internucleotide linkage at the 3′-end of said antisense region. In any of the embodiments of siNA molecules described herein, the antisense region can comprise about one to about five phosphorothioate internucleotide linkages at the 5′-end of said antisense region. In any of the embodiments of siNA molecules described herein, the 3′-terminal nucleotide overhangs of a siNA molecule of the invention can comprise ribonucleotides or deoxyribonucleotides that are chemically-modified at a nucleic acid sugar, base, or backbone. In any of the embodiments of siNA molecules described herein, the 3′-terminal nucleotide overhangs can comprise one or more universal base ribonucleotides. In any of the embodiments of siNA molecules described herein, the 3′-terminal nucleotide overhangs can comprise one or more acyclic nucleotides.

One embodiment of the invention provides an expression vector comprising a nucleic acid sequence encoding at least one siNA molecule of the invention in a manner that allows expression of the nucleic acid molecule. Another embodiment of the invention provides a mammalian cell comprising such an expression vector. The mammalian cell can be a human cell. The siNA molecule of the expression vector can comprise a sense region and an antisense region. The antisense region can comprise sequence complementary to a RNA or DNA sequence encoding a HDCA target and the sense region can comprise sequence complementary to the antisense region. The siNA molecule can comprise two distinct strands having complementary sense and antisense regions. The siNA molecule can comprise a single strand having complementary sense and antisense regions.

The chemically-modified internucleotide linkages having Formula I, for example, wherein any Z, W, X, and/or Y independently comprises a sulphur atom, can be present in one or both oligonucleotide strands of the siNA duplex, for example, in the sense strand, the antisense strand, or both strands. The siNA molecules of the invention can comprise one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) chemically-modified internucleotide linkages having Formula I at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the sense strand, the antisense strand, or both strands. For example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) chemically-modified internucleotide linkages having Formula I at the 5′-end of the sense strand, the antisense strand, or both strands. In another non-limiting example, an exemplary siNA molecule of the invention can comprise one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) pyrimidine nucleotides with chemically-modified internucleotide linkages having Formula I in the sense strand, the antisense strand, or both strands. In yet another non-limiting example, an exemplary siNA molecule of the invention can comprise one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) purine nucleotides with chemically-modified internucleotide linkages having Formula I in the sense strand, the antisense strand, or both strands. In another embodiment, a siNA molecule of the invention having internucleotide linkage(s) of Formula I also comprises a chemically-modified nucleotide or non-nucleotide having any of Formulae I-VII.

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule capable of mediating RNA interference (RNAi) inside a cell or reconstituted in vitro system, wherein the chemical modification comprises one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) nucleotides or non-nucleotides having Formula II:
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wherein each R3, R4, R5, R6, R7, R8, R10, R1 and R12 is independently H, OH, alkyl, substituted alkyl, alkaryl or aralkyl, F, Cl, Br, CN, CF3, OCF3, OCN, O-alkyl, S-alkyl, N-alkyl, O-alkenyl, S-alkenyl, N-alkenyl, SO-alkyl, alkyl-OSH, alkyl-OH, O-alkyl-OH, O-alkyl-SH, S-alkyl-OH, S-alkyl-SH, alkyl-S-alkyl, alkyl-O-alkyl, ONO2, NO2, N3, NH2, aminoalkyl, aminoacid, aminoacyl, ONH2, O-aminoalkyl, O-aminoacid, O-aminoacyl, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalklylamino, substituted silyl, or group having Formula I or II; R9 is O, S, CH2, S═O, CHF, or CF2, and B is a nucleosidic base such as adenine, guanine, uracil, cytosine, thymine, 2-aminoadenosine, 5-methylcytosine, 2,6-diaminopurine, or any other non-naturally occurring base that can be complementary or non-complementary to target RNA or a non-nucleosidic base such as phenyl, naphthyl, 3-nitropyrrole, 5-nitroindole, nebularine, pyridone, pyridinone, or any other non-naturally occurring universal base that can be complementary or non-complementary to target RNA. In one embodiment, R3 and/or R7 comprises a conjugate moiety and a linker (e.g., a nucleotide or non-nucleotide linker as described herein or otherwise known in the art). Non-limiting examples of conjugate moieties include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine.

The chemically-modified nucleotide or non-nucleotide of Formula II can be present in one or both oligonucleotide strands of the siNA duplex, for example in the sense strand, the antisense strand, or both strands. The siNA molecules of the invention can comprise one or more chemically-modified nucleotides or non-nucleotides of Formula II at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the sense strand, the antisense strand, or both strands. For example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) chemically-modified nucleotides or non-nucleotides of Formula II at the 5′-end of the sense strand, the antisense strand, or both strands. In anther non-limiting example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) chemically-modified nucleotides or non-nucleotides of Formula II at the 3′-end of the sense strand, the antisense strand, or both strands.

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule capable of mediating RNA interference (RNAi) inside a cell or reconstituted in vitro system, wherein the chemical modification comprises one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) nucleotides or non-nucleotides having Formula III:
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wherein each R3, R4, R5, R6, R7, R8, R10, R11 and R12 is independently H, OH, alkyl, substituted alkyl, alkaryl or aralkyl, F, Cl, Br, CN, CF3, OCF3, OCN, O-alkyl, S-alkyl, N-alkyl, O-alkenyl, S-alkenyl, N-alkenyl, SO-alkyl, alkyl-OSH, alkyl-OH, O-alkyl-OH, O-alkyl-SH, S-alkyl-OH, S-alkyl-SH, alkyl-S-alkyl, alkyl-O-alkyl, ONO2, NO2, N3, NH2, aminoalkyl, aminoacid, aminoacyl, ONH2, O-aminoalkyl, O-aminoacid, O-aminoacyl, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalklylamino, substituted silyl, or group having Formula I or II; R9 is O, S, CH2, S═O, CHF, or CF2, and B is a nucleosidic base such as adenine, guanine, uracil, cytosine, thymine, 2-aminoadenosine, 5-methylcytosine, 2,6-diaminopurine, or any other non-naturally occurring base that can be employed to be complementary or non-complementary to target RNA or a non-nucleosidic base such as phenyl, naphthyl, 3-nitropyrrole, 5-nitroindole, nebularine, pyridone, pyridinone, or any other non-naturally occurring universal base that can be complementary or non-complementary to target RNA. In one embodiment, R3 and/or R7 comprises a conjugate moiety and a linker (e.g., a nucleotide or non-nucleotide linker as described herein or otherwise known in the art). Non-limiting examples of conjugate moieties include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine.

The chemically-modified nucleotide or non-nucleotide of Formula III can be present in one or both oligonucleotide strands of the siNA duplex, for example, in the sense strand, the antisense strand, or both strands. The siNA molecules of the invention can comprise one or more chemically-modified nucleotides or non-nucleotides of Formula III at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the sense strand, the antisense strand, or both strands. For example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) chemically-modified nucleotide(s) or non-nucleotide(s) of Formula III at the 5′-end of the sense strand, the antisense strand, or both strands. In anther non-limiting example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) chemically-modified nucleotide or non-nucleotide of Formula III at the 3′-end of the sense strand, the antisense strand, or both strands.

In another embodiment, a siNA molecule of the invention comprises a nucleotide having Formula II or III, wherein the nucleotide having Formula II or III is in an inverted configuration. For example, the nucleotide having Formula II or III is connected to the siNA construct in a 3′-3′,3′-2′,2′-3′, or 5′-5′ configuration, such as at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of one or both siNA strands.

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule capable of mediating RNA interference (RNAi) inside a cell or reconstituted in vitro system, wherein the chemical modification comprises a 5′-terminal phosphate group having Formula IV:
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wherein each X and Y is independently O, S, N, alkyl, substituted alkyl, or alkylhalo; wherein each Z and W is independently O, S, N, alkyl, substituted alkyl, O-alkyl, S-alkyl, alkaryl, aralkyl, alkylhalo, or acetyl; and wherein W, X, Y and Z are not all O.

In one embodiment, the invention features a siNA molecule having a 5′-terminal phosphate group having Formula IV on the target-complementary strand, for example, a strand complementary to a target RNA, wherein the siNA molecule comprises an all RNA siNA molecule. In another embodiment, the invention features a siNA molecule having a 5′-terminal phosphate group having Formula IV on the target-complementary strand wherein the siNA molecule also comprises about 1 to about 3 (e.g., about 1, 2, or 3) nucleotide 3′-terminal nucleotide overhangs having about 1 to about 4 (e.g., about 1, 2, 3, or 4) deoxyribonucleotides on the 3′-end of one or both strands. In another embodiment, a 5′-terminal phosphate group having Formula IV is present on the target-complementary strand of a siNA molecule of the invention, for example a siNA molecule having chemical modifications having any of Formulae I-VII.

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule capable of mediating RNA interference (RNAi) inside a cell or reconstituted in vitro system, wherein the chemical modification comprises one or more phosphorothioate internucleotide linkages. For example, in a non-limiting example, the invention features a chemically-modified short interfering nucleic acid (siNA) having about 1, 2, 3, 4, 5, 6, 7, 8 or more phosphorothioate internucleotide linkages in one siNA strand. In yet another embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) individually having about 1, 2, 3, 4, 5, 6, 7, 8 or more phosphorothioate internucleotide linkages in both siNA strands. The phosphorothioate internucleotide linkages can be present in one or both oligonucleotide strands of the siNA duplex, for example in the sense strand, the antisense strand, or both strands. The siNA molecules of the invention can comprise one or more phosphorothioate internucleotide linkages at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand, the antisense strand, or both strands. For example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) consecutive phosphorothioate internucleotide linkages at the 5′-end of the sense strand, the antisense strand, or both strands. In another non-limiting example, an exemplary siNA molecule of the invention can comprise one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) pyrimidine phosphorothioate internucleotide linkages in the sense strand, the antisense strand, or both strands. In yet another non-limiting example, an exemplary siNA molecule of the invention can comprise one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) purine phosphorothioate internucleotide linkages in the sense strand, the antisense strand, or both strands.

Each strand of the double stranded siNA molecule can have one or more chemical modifications such that each strand comprises a different pattern of chemical modifications. Several non-limiting examples of modification schemes that could give rise to different patterns of modifications are provided herein.

In one embodiment, the invention features a siNA molecule, wherein the sense strand comprises one or more, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy and/or about one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and wherein the antisense strand comprises about 1 to about 10 or more, specifically about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In another embodiment, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more, pyrimidine nucleotides of the sense and/or antisense siNA strand are chemically-modified with 2′-deoxy, 2′-O-methyl, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or 2′-deoxy-2′-fluoro nucleotides, with or without one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more, phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends, being present in the same or different strand.

In another embodiment, the invention features a siNA molecule, wherein the sense strand comprises about 1 to about 5, specifically about 1, 2, 3, 4, or 5 phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and wherein the antisense strand comprises about 1 to about 5 or more, specifically about 1, 2, 3, 4, 5, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In another embodiment, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more, pyrimidine nucleotides of the sense and/or antisense siNA strand are chemically-modified with 2′-deoxy, 2′-O-methyl, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or 2′-deoxy-2′-fluoro nucleotides, with or without about 1 to about 5 or more, for example about 1, 2, 3, 4, 5, or more phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends, being present in the same or different strand.

In one embodiment, the invention features a siNA molecule, wherein the antisense strand comprises one or more, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more phosphorothioate internucleotide linkages, and/or about one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and wherein the antisense strand comprises about 1 to about 10 or more, specifically about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In another embodiment, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more pyrimidine nucleotides of the sense and/or antisense siNA strand are chemically-modified with 2′-deoxy, 2′-O-methyl, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or 2′-deoxy-2′-fluoro nucleotides, with or without one or more, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends, being present in the same or different strand.

In another embodiment, the invention features a siNA molecule, wherein the antisense strand comprises about 1 to about 5 or more, specifically about 1, 2, 3, 4, 5 or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and wherein the antisense strand comprises about 1 to about 5 or more, specifically about 1, 2, 3, 4, 5 or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In another embodiment, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more pyrimidine nucleotides of the sense and/or antisense siNA strand are chemically-modified with 2′-deoxy, 2′-O-methyl, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or 2′-deoxy-2′-fluoro nucleotides, with or without about 1 to about 5, for example about 1, 2, 3, 4, 5 or more phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends, being present in the same or different strand.

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule having about 1 to about 5 or more (specifically about 1, 2, 3, 4, 5 or more) phosphorothioate internucleotide linkages in each strand of the siNA molecule.

In another embodiment, the invention features a siNA molecule comprising 2′-5′ internucleotide linkages. The 2′-5′ internucleotide linkage(s) can be at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of one or both siNA sequence strands. In addition, the 2′-5′ internucleotide linkage(s) can be present at various other positions within one or both siNA sequence strands, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more including every internucleotide linkage of a pyrimidine nucleotide in one or both strands of the siNA molecule can comprise a 2′-5′ internucleotide linkage, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more including every internucleotide linkage of a purine nucleotide in one or both strands of the siNA molecule can comprise a 2′-5′ internucleotide linkage.

In another embodiment, a chemically-modified siNA molecule of the invention comprises a duplex having two strands, one or both of which can be chemically-modified, wherein each strand is independently about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length, wherein the duplex has about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) base pairs, and wherein the chemical modification comprises a structure having any of Formulae I-VII. For example, an exemplary chemically-modified siNA molecule of the invention comprises a duplex having two strands, one or both of which can be chemically-modified with a chemical modification having any of Formulae I-VII or any combination thereof, wherein each strand consists of about 21 nucleotides, each having a 2-nucleotide 3′-terminal nucleotide overhang, and wherein the duplex has about 19 base pairs. In another embodiment, a siNA molecule of the invention comprises a single stranded hairpin structure, wherein the siNA is about 36 to about 70 (e.g., about 36, 40, 45, 50, 55, 60, 65, or 70) nucleotides in length having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) base pairs, and wherein the siNA can include a chemical modification comprising a structure having any of Formulae I-VII or any combination thereof. For example, an exemplary chemically-modified siNA molecule of the invention comprises a linear oligonucleotide having about 42 to about 50 (e.g., about 42, 43, 44, 45, 46, 47, 48, 49, or 50) nucleotides that is chemically-modified with a chemical modification having any of Formulae I-VII or any combination thereof, wherein the linear oligonucleotide forms a hairpin structure having about 19 to about 21 (e.g., 19, 20, or 21) base pairs and a 2-nucleotide 3′-terminal nucleotide overhang. In another embodiment, a linear hairpin siNA molecule of the invention contains a stem loop motif, wherein the loop portion of the siNA molecule is biodegradable. For example, a linear hairpin siNA molecule of the invention is designed such that degradation of the loop portion of the siNA molecule in vivo can generate a double-stranded siNA molecule with 3′-terminal overhangs, such as 3′-terminal nucleotide overhangs comprising about 2 nucleotides.

In another embodiment, a siNA molecule of the invention comprises a hairpin structure, wherein the siNA is about 25 to about 50 (e.g., about 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50) nucleotides in length having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) base pairs, and wherein the siNA can include one or more chemical modifications comprising a structure having any of Formulae I-VII or any combination thereof. For example, an exemplary chemically-modified siNA molecule of the invention comprises a linear oligonucleotide having about 25 to about 35 (e.g., about 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) nucleotides that is chemically-modified with one or more chemical modifications having any of Formulae I-VII or any combination thereof, wherein the linear oligonucleotide forms a hairpin structure having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) base pairs and a 5′-terminal phosphate group that can be chemically modified as described herein (for example a 5′-terminal phosphate group having Formula IV). In another embodiment, a linear hairpin siNA molecule of the invention contains a stem loop motif, wherein the loop portion of the siNA molecule is biodegradable. In one embodiment, a linear hairpin siNA molecule of the invention comprises a loop portion comprising a non-nucleotide linker.

In another embodiment, a siNA molecule of the invention comprises an asymmetric hairpin structure, wherein the siNA is about 25 to about 50 (e.g., about 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50) nucleotides in length having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) base pairs, and wherein the siNA can include one or more chemical modifications comprising a structure having any of Formulae I-VII or any combination thereof. For example, an exemplary chemically-modified siNA molecule of the invention comprises a linear oligonucleotide having about 25 to about 35 (e.g., about 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) nucleotides that is chemically-modified with one or more chemical modifications having any of Formulae I-VII or any combination thereof, wherein the linear oligonucleotide forms an asymmetric hairpin structure having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) base pairs and a 5′-terminal phosphate group that can be chemically modified as described herein (for example a 5′-terminal phosphate group having Formula IV). In one embodiment, an asymmetric hairpin siNA molecule of the invention contains a stem loop motif, wherein the loop portion of the siNA molecule is biodegradable. In another embodiment, an asymmetric hairpin siNA molecule of the invention comprises a loop portion comprising a non-nucleotide linker.

In another embodiment, a siNA molecule of the invention comprises an asymmetric double stranded structure having separate polynucleotide strands comprising sense and antisense regions, wherein the antisense region is about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length, wherein the sense region is about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) nucleotides in length, wherein the sense region and the antisense region have at least 3 complementary nucleotides, and wherein the siNA can include one or more chemical modifications comprising a structure having any of Formulae I-VII or any combination thereof. For example, an exemplary chemically-modified siNA molecule of the invention comprises an asymmetric double stranded structure having separate polynucleotide strands comprising sense and antisense regions, wherein the antisense region is about 18 to about 23 (e.g., about 18, 19, 20, 21, 22, or 23) nucleotides in length and wherein the sense region is about 3 to about 15 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15) nucleotides in length, wherein the sense region the antisense region have at least 3 complementary nucleotides, and wherein the siNA can include one or more chemical modifications comprising a structure having any of Formulae I-VII or any combination thereof. In another embodiment, the asymmetric double stranded siNA molecule can also have a 5′-terminal phosphate group that can be chemically modified as described herein (for example a 5′-terminal phosphate group having Formula IV).

In another embodiment, a siNA molecule of the invention comprises a circular nucleic acid molecule, wherein the siNA is about 38 to about 70 (e.g., about 38, 40, 45, 50, 55, 60, 65, or 70) nucleotides in length having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) base pairs, and wherein the siNA can include a chemical modification, which comprises a structure having any of Formulae I-VII or any combination thereof. For example, an exemplary chemically-modified siNA molecule of the invention comprises a circular oligonucleotide having about 42 to about 50 (e.g., about 42, 43, 44, 45, 46, 47, 48, 49, or 50) nucleotides that is chemically-modified with a chemical modification having any of Formulae I-VII or any combination thereof, wherein the circular oligonucleotide forms a dumbbell shaped structure having about 19 base pairs and 2 loops.

In another embodiment, a circular siNA molecule of the invention contains two loop motifs, wherein one or both loop portions of the siNA molecule is biodegradable. For example, a circular siNA molecule of the invention is designed such that degradation of the loop portions of the siNA molecule in vivo can generate a double-stranded siNA molecule with 3′-terminal overhangs, such as 3′-terminal nucleotide overhangs comprising about 2 nucleotides.

In one embodiment, a siNA molecule of the invention comprises at least one (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) abasic moiety, for example a compound having Formula V:
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wherein each R3, R4, R5, R6, R7, R8, R10, R11, R12, and R13 is independently H, OH, alkyl, substituted alkyl, alkaryl or aralkyl, F, Cl, Br, CN, CF3, OCF3, OCN, O-alkyl, S-alkyl, N-alkyl, O-alkenyl, S-alkenyl, N-alkenyl, SO-alkyl, alkyl-OSH, alkyl-OH, O-alkyl-OH, O-alkyl-SH, S-alkyl-OH, S-alkyl-SH, alkyl-S-alkyl, alkyl-O-alkyl, ONO2, NO2, N3, NH2, aminoalkyl, aminoacid, aminoacyl, ONH2, O-aminoalkyl, O-aminoacid, O-aminoacyl, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalklylamino, substituted silyl, or group having Formula I or II; R9 is O, S, CH2, S═O, CHF, or CF2. In one embodiment, R3 and/or R7 comprises a conjugate moiety and a linker (e.g., a nucleotide or non-nucleotide linker as described herein or otherwise known in the art). Non-limiting examples of conjugate moieties include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine.

In one embodiment, a siNA molecule of the invention comprises at least one (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) inverted abasic moiety, for example a compound having Formula VI:
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wherein each R3, R4, R5, R6, R7, R8, R10, R11, R12, and R13 is independently H, OH, alkyl, substituted alkyl, alkaryl or aralkyl, F, Cl, Br, CN, CF3, OCF3, OCN, O-alkyl, S-alkyl, N-alkyl, O-alkenyl, S-alkenyl, N-alkenyl, SO-alkyl, alkyl-OSH, alkyl-OH, O-alkyl-OH, O-alkyl-SH, S-alkyl-OH, S-alkyl-SH, alkyl-S-alkyl, alkyl-O-alkyl, ONO2, NO2, N3, NH2, aminoalkyl, aminoacid, aminoacyl, ONH2, O-aminoalkyl, O-aminoacid, O-aminoacyl, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalklylamino, substituted silyl, or group having Formula I or II; R9 is O, S, CH2, S═O, CHF, or CF2, and either R2, R3, R8 or R13 serve as points of attachment to the siNA molecule of the invention. In one embodiment, R3 and/or R7 comprises a conjugate moiety and a linker (e.g., a nucleotide or non-nucleotide linker as described herein or otherwise known in the art). Non-limiting examples of conjugate moieties include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine.

In another embodiment, a siNA molecule of the invention comprises at least one (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) substituted polyalkyl moieties, for example a compound having Formula VII:
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wherein each n is independently an integer from 1 to 12, each R1, R2 and R3 is independently H, OH, alkyl, substituted alkyl, alkaryl or aralkyl, F, Cl, Br, CN, CF3, OCF3, OCN, O-alkyl, S-alkyl, N-alkyl, O-alkenyl, S-alkenyl, N-alkenyl, SO-alkyl, alkyl-OSH, alkyl-OH, O-alkyl-OH, O-alkyl-SH, S-alkyl-OH, S-alkyl-SH, alkyl-S-alkyl, alkyl-O-alkyl, ONO2, NO2, N3, NH2, aminoalkyl, aminoacid, aminoacyl, ONH2, O-aminoalkyl, O-aminoacid, O-aminoacyl, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalklylamino, substituted silyl, or a group having Formula I, and R1, R2 or R3 serves as points of attachment to the siNA molecule of the invention. In one embodiment, R3 and/or R1 comprises a conjugate moiety and a linker (e.g., a nucleotide or non-nucleotide linker as described herein or otherwise known in the art). Non-limiting examples of conjugate moieties include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine.

By “ZIP code” sequences is meant, any peptide or protein sequence that is involved in cellular topogenic signaling mediated transport (see for example Ray et al., 2004, Science, 306(1501): 1505)

Each nucleotide within the double stranded siNA molecule can independently have a chemical modification comprising the structure of any of Formulae I-VIII. Thus, in one embodiment, one or more nucleotide positions of a siNA molecule of the invention comprises a chemical modification having structure of any of Formulae I-VII or any other modification herein. In one embodiment, each nucleotide position of a siNA molecule of the invention comprises a chemical modification having structure of any of Formulae I-VII or any other modification herein.

In one embodiment, one or more nucleotide positions of one or both strands of a double stranded siNA molecule of the invention comprises a chemical modification having structure of any of Formulae I-VII or any other modification herein. In one embodiment, each nucleotide position of one or both strands of a double stranded siNA molecule of the invention comprises a chemical modification having structure of any of Formulae I-VII or any other modification herein.

In another embodiment, the invention features a compound having Formula VII, wherein R1 and R2 are hydroxyl (OH) groups, n=1, and R3 comprises O and is the point of attachment to the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of one or both strands of a double-stranded siNA molecule of the invention or to a single-stranded siNA molecule of the invention. This modification is referred to herein as “glyceryl” (for example modification 6 in FIG. 10).

In another embodiment, a chemically modified nucleoside or non-nucleoside (e.g. a moiety having any of Formula V, VI or VII) of the invention is at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of a siNA molecule of the invention. For example, chemically modified nucleoside or non-nucleoside (e.g., a moiety having Formula V, VI or VII) can be present at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the antisense strand, the sense strand, or both antisense and sense strands of the siNA molecule. In one embodiment, the chemically modified nucleoside or non-nucleoside (e.g., a moiety having Formula V, VI or VII) is present at the 5′-end and 3′-end of the sense strand and the 3′-end of the antisense strand of a double stranded siNA molecule of the invention. In one embodiment, the chemically modified nucleoside or non-nucleoside (e.g., a moiety having Formula V, VI or VII) is present at the terminal position of the 5′-end and 3′-end of the sense strand and the 3′-end of the antisense strand of a double stranded siNA molecule of the invention. In one embodiment, the chemically modified nucleoside or non-nucleoside (e.g., a moiety having Formula V, VI or VII) is present at the two terminal positions of the 5′-end and 3′-end of the sense strand and the 3′-end of the antisense strand of a double stranded siNA molecule of the invention. In one embodiment, the chemically modified nucleoside or non-nucleoside (e.g., a moiety having Formula V, VI or VII) is present at the penultimate position of the 5′-end and 3′-end of the sense strand and the 3′-end of the antisense strand of a double stranded siNA molecule of the invention. In addition, a moiety having Formula VII can be present at the 3′-end or the 5′-end of a hairpin siNA molecule as described herein.

In another embodiment, a siNA molecule of the invention comprises an abasic residue having Formula V or VI, wherein the abasic residue having Formula VI or VI is connected to the siNA construct in a 3′-3′,3′-2′,2′-3′, or 5′-5′ configuration, such as at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of one or both siNA strands.

In one embodiment, a siNA molecule of the invention comprises one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) locked nucleic acid (LNA) nucleotides, for example, at the 5′-end, the 3′-end, both of the 5′ and 3′-ends, or any combination thereof, of the siNA molecule.

In one embodiment, a siNA molecule of the invention comprises one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) 4′-thio nucleotides, for example, at the 5′-end, the 3′-end, both of the 5′ and 3′-ends, or any combination thereof, of the siNA molecule.

In another embodiment, a siNA molecule of the invention comprises one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) acyclic nucleotides, for example, at the 5′-end, the 3′-end, both of the 5′ and 3′-ends, or any combination thereof, of the siNA molecule.

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising a sense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the sense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any (e.g., one or more or all) purine nucleotides present in the sense region are 2′-deoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-deoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-deoxy purine nucleotides), wherein any nucleotides comprising a 3′-terminal nucleotide overhang that are present in said sense region are 2′-deoxy nucleotides.

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising a sense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the sense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any (e.g., one or more or all) purine nucleotides present in the sense region are 2′-O-methyl purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides).

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising a sense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the sense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), wherein any (e.g., one or more or all) purine nucleotides present in the sense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides), and wherein any nucleotides comprising a 3′-terminal nucleotide overhang that are present in said sense region are 2′-deoxy nucleotides.

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising an antisense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the antisense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g. wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any (e.g., one or more or all) purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides).

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising an antisense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the antisense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), wherein any (e.g., one or more or all) purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides), and wherein any nucleotides comprising a 3′-terminal nucleotide overhang that are present in said antisense region are 2′-deoxy nucleotides.

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising an antisense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the antisense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any (e.g., one or more or all) purine nucleotides present in the antisense region are 2′-deoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-deoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-deoxy purine nucleotides).

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising an antisense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the antisense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any (e.g., one or more or all) purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides).

In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention capable of mediating RNA interference (RNAi) inside a cell or reconstituted in vitro system comprising a sense region, wherein one or more pyrimidine nucleotides present in the sense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and one or more purine nucleotides present in the sense region are 2′-deoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-deoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-deoxy purine nucleotides), and an antisense region, wherein one or more pyrimidine nucleotides present in the antisense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and one or more purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides). The sense region and/or the antisense region can have a terminal cap modification, such as any modification described herein or shown in FIG. 10, that is optionally present at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the sense and/or antisense sequence. The sense and/or antisense region can optionally further comprise a 3′-terminal nucleotide overhang having about 1 to about 4 (e.g., about 1, 2, 3, or 4) 2′-deoxynucleotides. The overhang nucleotides can further comprise one or more (e.g., about 1, 2, 3, 4 or more) phosphorothioate, phosphonoacetate, and/or thiophosphonoacetate internucleotide linkages. Non-limiting examples of these chemically-modified siNAs are shown in FIGS. 4 and 5 and Tables III and IV herein. In any of these described embodiments, the purine nucleotides present in the sense region are alternatively 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides) and one or more purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides). Also, in any of these embodiments, one or more purine nucleotides present in the sense region are alternatively purine ribonucleotides (e.g., wherein all purine nucleotides are purine ribonucleotides or alternately a plurality of purine nucleotides are purine ribonucleotides) and any purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides). Additionally, in any of these embodiments, one or more purine nucleotides present in the sense region and/or present in the antisense region are alternatively selected from the group consisting of 2′-deoxy nucleotides, locked nucleic acid (LNA) nucleotides, 2′-methoxyethyl nucleotides, 4′-thionucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides and 2′-O-methyl nucleotides (e.g., wherein all purine nucleotides are selected from the group consisting of 2′-deoxy nucleotides, locked nucleic acid (LNA) nucleotides, 2′-methoxyethyl nucleotides, 4′-thionucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides and 2′-O-methyl nucleotides or alternately a plurality of purine nucleotides are selected from the group consisting of 2′-deoxy nucleotides, locked nucleic acid (LNA) nucleotides, 2′-methoxyethyl nucleotides, 4′-thionucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides and 2′-O-methyl nucleotides).

In another embodiment, any modified nucleotides present in the siNA molecules of the invention, preferably in the antisense strand of the siNA molecules of the invention, but also optionally in the sense and/or both antisense and sense strands, comprise modified nucleotides having properties or characteristics similar to naturally occurring ribonucleotides. For example, the invention features siNA molecules including modified nucleotides having a Northern conformation (e.g., Northern pseudorotation cycle, see for example Saenger, Principles of Nucleic Acid Structure, Springer-Verlag ed., 1984). Such nucleotides having a Northern conformation are generally considered to be “ribo-like” as they have a C3′-endo sugar pucker conformation. As such, chemically modified nucleotides present in the siNA molecules of the invention, preferably in the antisense strand of the siNA molecules of the invention, but also optionally in the sense and/or both antisense and sense strands, are resistant to nuclease degradation while at the same time maintaining the capacity to mediate RNAi. Non-limiting examples of nucleotides having a northern configuration include locked nucleic acid (LNA) nucleotides (e.g., 2′-O, 4′-C-methylene-(D-ribofuranosyl) nucleotides); 2′-methoxyethoxy (MOE) nucleotides; 2′-methyl-thio-ethyl, 2′-deoxy-2′-fluoro nucleotides, 2′-deoxy-2′-chloro nucleotides, 2′-azido nucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides, 4′-thio nucleotides and 2′-O-methyl nucleotides.

In one embodiment, the sense strand of a double stranded siNA molecule of the invention comprises a terminal cap moiety, (see for example FIG. 10) such as an inverted deoxyabaisc moiety, at the 3′-end, 5′-end, or both 3′ and 5′-ends of the sense strand.

In one embodiment, the invention features a chemically-modified short interfering nucleic acid molecule (siNA) capable of mediating RNA interference (RNAi) inside a cell or reconstituted in vitro system, wherein the chemical modification comprises a conjugate covalently attached to the chemically-modified siNA molecule. Non-limiting examples of conjugates contemplated by the invention include conjugates and ligands described in Vargeese et al., U.S. Ser. No. 10/427,160, filed Apr. 30, 2003, incorporated by reference herein in its entirety, including the drawings. In another embodiment, the conjugate is covalently attached to the chemically-modified siNA molecule via a biodegradable linker. In one embodiment, the conjugate molecule is attached at the 3′-end of either the sense strand, the antisense strand, or both strands of the chemically-modified siNA molecule. In another embodiment, the conjugate molecule is attached at the 5′-end of either the sense strand, the antisense strand, or both strands of the chemically-modified siNA molecule. In yet another embodiment, the conjugate molecule is attached both the 3′-end and 5′-end of either the sense strand, the antisense strand, or both strands of the chemically-modified siNA molecule, or any combination thereof. In one embodiment, a conjugate molecule of the invention comprises a molecule that facilitates delivery of a chemically-modified siNA molecule into a biological system, such as a cell. In another embodiment, the conjugate molecule attached to the chemically-modified siNA molecule is a ligand for a cellular receptor, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine. Examples of specific conjugate molecules contemplated by the instant invention that can be attached to chemically-modified siNA molecules are described in Vargeese et al., U.S. Ser. No. 10/201,394, filed Jul. 22, 2002 incorporated by reference herein. The type of conjugates used and the extent of conjugation of siNA molecules of the invention can be evaluated for improved pharmacokinetic profiles, bioavailability, and/or stability of siNA constructs while at the same time maintaining the ability of the siNA to mediate RNAi activity. As such, one skilled in the art can screen siNA constructs that are modified with various conjugates to determine whether the siNA conjugate complex possesses improved properties while maintaining the ability to mediate RNAi, for example in animal models as are generally known in the art.

In one embodiment, the invention features a short interfering nucleic acid (siNA) molecule of the invention, wherein the siNA further comprises a nucleotide, non-nucleotide, or mixed nucleotide/non-nucleotide linker that joins the sense region of the siNA to the antisense region of the siNA. In one embodiment, a nucleotide, non-nucleotide, or mixed nucleotide/non-nucleotide linker is used, for example, to attach a conjugate moiety to the siNA. In one embodiment, a nucleotide linker of the invention can be a linker of >2 nucleotides in length, for example about 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides in length. In another embodiment, the nucleotide linker can be a nucleic acid aptamer. By “aptamer” or “nucleic acid aptamer” as used herein is meant a nucleic acid molecule that binds specifically to a target molecule wherein the nucleic acid molecule has sequence that comprises a sequence recognized by the target molecule in its natural setting. Alternately, an aptamer can be a nucleic acid molecule that binds to a target molecule where the target molecule does not naturally bind to a nucleic acid. The target molecule can be any molecule of interest. For example, the aptamer can be used to bind to a ligand-binding domain of a protein, thereby preventing interaction of the naturally occurring ligand with the protein. This is a non-limiting example and those in the art will recognize that other embodiments can be readily generated using techniques generally known in the art. (See, for example, Gold et al., 1995, Annu. Rev. Biochem., 64, 763; Brody and Gold, 2000, J. Biotechnol., 74, 5; Sun, 2000, Curr. Opin. Mol. Ther., 2, 100; Kusser, 2000, J. Biotechnol., 74, 27; Hermann and Patel, 2000, Science, 287, 820; and Jayasena, 1999, Clinical Chemistry, 45, 1628.)

In yet another embodiment, a non-nucleotide linker of the invention comprises abasic nucleotide, polyether, polyamine, polyamide, peptide, carbohydrate, lipid, polyhydrocarbon, or other polymeric compounds (e.g. polyethylene glycols such as those having between 2 and 100 ethylene glycol units). Specific examples include those described by Seela and Kaiser, Nucleic Acids Res. 1990, 18:6353 and Nucleic Acids Res. 1987, 15:3113; Cload and Schepartz, J. Am. Chem. Soc. 1991, 113:6324; Richardson and Schepartz, J. Am. Chem. Soc. 1991, 113:5109; Ma et al., Nucleic Acids Res. 1993, 21:2585 and Biochemistry 1993, 32:1751; Durand et al., Nucleic Acids Res. 1990, 18:6353; McCurdy et al., Nucleosides & Nucleotides 1991, 10:287; Jschke et al., Tetrahedron Lett. 1993, 34:301; Ono et al., Biochemistry 1991, 30:9914; Arnold et al., International Publication No. WO 89/02439; Usman et al., International Publication No. WO 95/06731; Dudycz et al., International Publication No. WO 95/11910 and Ferentz and Verdine, J. Am. Chem. Soc. 1991, 113:4000, all hereby incorporated by reference herein. A “non-nucleotide” further means any group or compound that can be incorporated into a nucleic acid chain in the place of one or more nucleotide units, including either sugar and/or phosphate substitutions, and allows the remaining bases to exhibit their enzymatic activity. The group or compound can be abasic in that it does not contain a commonly recognized nucleotide base, such as adenosine, guanine, cytosine, uracil or thymine, for example at the C1 position of the sugar.

In one embodiment, the invention features a short interfering nucleic acid (siNA) molecule capable of mediating RNA interference (RNAi) inside a cell or reconstituted in vitro system, wherein one or both strands of the siNA molecule that are assembled from two separate oligonucleotides do not comprise any ribonucleotides. For example, a siNA molecule can be assembled from a single oligonculeotide where the sense and antisense regions of the siNA comprise separate oligonucleotides that do not have any ribonucleotides (e.g., nucleotides having a 2′-OH group) present in the oligonucleotides. In another example, a siNA molecule can be assembled from a single oligonculeotide where the sense and antisense regions of the siNA are linked or circularized by a nucleotide or non-nucleotide linker as described herein, wherein the oligonucleotide does not have any ribonucleotides (e.g., nucleotides having a 2′-OH group) present in the oligonucleotide. Applicant has surprisingly found that the presense of ribonucleotides (e.g., nucleotides having a 2′-hydroxyl group) within the siNA molecule is not required or essential to support RNAi activity. As such, in one embodiment, all positions within the siNA can include chemically modified nucleotides and/or non-nucleotides such as nucleotides and or non-nucleotides having Formula I, II, III, IV, V, VI, or VII or any combination thereof to the extent that the ability of the siNA molecule to support RNAi activity in a cell is maintained.

In one embodiment, a siNA molecule of the invention is a single stranded siNA molecule that mediates RNAi activity in a cell or reconstituted in vitro system comprising a single stranded polynucleotide having complementarity to a target HDAC nucleic acid sequence, wherein one or more pyrimidine nucleotides present in the siNA are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides), and a terminal cap modification, such as any modification described herein or shown in FIG. 10, that is optionally present at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the antisense sequence. The siNA optionally further comprises about 1 to about 4 or more (e.g., about 1, 2, 3, 4 or more) terminal 2′-deoxynucleotides at the 3′-end of the siNA molecule, wherein the terminal nucleotides can further comprise one or more (e.g., 1, 2, 3, 4 or more) phosphorothioate, phosphonoacetate, and/or thiophosphonoacetate internucleotide linkages, and wherein the siNA optionally further comprises a terminal phosphate group, such as a 5′-terminal phosphate group. In any of these embodiments, any purine nucleotides present in the antisense region are alternatively 2′-deoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-deoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-deoxy purine nucleotides). Also, in any of these embodiments, any purine nucleotides present in the siNA (i.e., purine nucleotides present in the sense and/or antisense region) can alternatively be locked nucleic acid (LNA) nucleotides (e.g., wherein all purine nucleotides are LNA nucleotides or alternately a plurality of purine nucleotides are LNA nucleotides). Also, in any of these embodiments, any purine nucleotides present in the siNA are alternatively 2′-methoxyethyl purine nucleotides (e.g., wherein all purine nucleotides are 2′-methoxyethyl purine nucleotides or alternately a plurality of purine nucleotides are 2′-methoxyethyl purine nucleotides). In another embodiment, any modified nucleotides present in the single stranded siNA molecules of the invention comprise modified nucleotides having properties or characteristics similar to naturally occurring ribonucleotides. For example, the invention features siNA molecules including modified nucleotides having a Northern conformation (e.g., Northern pseudorotation cycle, see for example Saenger, Principles of Nucleic Acid Structure, Springer-Verlag ed., 1984). As such, chemically modified nucleotides present in the single stranded siNA molecules of the invention are preferably resistant to nuclease degradation while at the same time maintaining the capacity to mediate RNAi.

In one embodiment, a siNA molecule of the invention comprises chemically modified nucleotides or non-nucleotides (e.g., having any modification described herein, including any of Formulae I-VII, such as 2′-deoxy, 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy or 2′-O-methyl nucleotides) at alternating positions within one or more strands or regions of the siNA molecule. For example, such chemical modifications can be introduced at every other position of a RNA based siNA molecule, starting at either the first or second nucleotide from the 3′-end or 5′-end of the siNA. In a non-limiting example, a double stranded siNA molecule of the invention in which each strand of the siNA is 21 nucleotides in length is featured wherein positions 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 and 21 of each strand are chemically modified (e.g., with compounds having any of Formulae I-VII, such as such as 2′-deoxy, 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy or 2′-O-methyl nucleotides). In another non-limiting example, a double stranded siNA molecule of the invention in which each strand of the siNA is 21 nucleotides in length is featured wherein positions 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 of each strand are chemically modified (e.g., with compounds having any of Formulae I-VII, such as such as 2′-deoxy, 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy or 2′-O-methyl nucleotides). In one embodiment, one strand of the double stranded siNA molecule comprises chemical modifications at positions 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 and chemical modifications at positions 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 and 21. Such siNA molecules can further comprise terminal cap moieties and/or backbone modifications as described herein.

In one embodiment, the invention features a method for modulating the expression of a target HDAC gene within a cell comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified or unmodified, wherein one of the siNA strands comprises a sequence complementary to RNA of the target HDAC gene; and (b) introducing the siNA molecule into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in the cell.

In one embodiment, the invention features a method for modulating the expression of a target HDAC gene within a cell comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified or unmodified, wherein one of the siNA strands comprises a sequence complementary to RNA of the target HDAC gene and wherein the sense strand sequence of the siNA comprises a sequence identical or substantially similar to the sequence of the target HDAC RNA; and (b) introducing the siNA molecule into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in the cell.

In another embodiment, the invention features a method for modulating the expression of more than one target HDAC gene within a cell comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified or unmodified, wherein one of the siNA strands comprises a sequence complementary to RNA of the target HDAC genes; and (b) introducing the siNA molecules into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC genes in the cell.

In another embodiment, the invention features a method for modulating the expression of two or more target HDAC genes within a cell comprising: (a) synthesizing one or more siNA molecules of the invention, which can be chemically-modified or unmodified, wherein the siNA strands comprise sequences complementary to RNA of the target HDAC genes and wherein the sense strand sequences of the siNAs comprise sequences identical or substantially similar to the sequences of the target HDAC RNAs; and (b) introducing the siNA molecules into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC genes in the cell.

In another embodiment, the invention features a method for modulating the expression of more than one target HDAC gene within a cell comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified or unmodified, wherein one of the siNA strands comprises a sequence complementary to RNA of the target HDAC gene and wherein the sense strand sequence of the siNA comprises a sequence identical or substantially similar to the sequences of the target HDAC RNAs; and (b) introducing the siNA molecule into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC genes in the cell.

In another embodiment, the invention features a method for modulating the expression of a target HDAC gene within a cell comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified or unmodified, wherein one of the siNA strands comprises a sequence complementary to RNA of the target HDAC gene, wherein the sense strand sequence of the siNA comprises a sequence identical or substantially similar to the sequences of the target HDAC RNA; and (b) introducing the siNA molecule into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in the cell.

In one embodiment, siNA molecules of the invention are used as reagents in ex vivo applications. For example, siNA reagents are introduced into tissue or cells that are transplanted into a subject for therapeutic effect. The cells and/or tissue can be derived from an organism or subject that later receives the explant, or can be derived from another organism or subject prior to transplantation. The siNA molecules can be used to modulate the expression of one or more genes in the cells or tissue, such that the cells or tissue obtain a desired phenotype or are able to perform a function when transplanted in vivo. In one embodiment, certain target HDAC cells from a patient are extracted. These extracted cells are contacted with siNAs target HDACing a specific nucleotide sequence within the cells under conditions suitable for uptake of the siNAs by these cells (e.g. using delivery reagents such as cationic lipids, liposomes and the like or using techniques such as electroporation to facilitate the delivery of siNAs into cells). The cells are then reintroduced back into the same patient or other patients.

In one embodiment, the invention features a method of modulating the expression of a target HDAC gene in a tissue explant comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein one of the siNA strands comprises a sequence complementary to RNA of the target HDAC gene; and (b) introducing the siNA molecule into a cell of the tissue explant derived from a particular organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in the tissue explant. In another embodiment, the method further comprises introducing the tissue explant back into the organism the tissue was derived from or into another organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in that organism.

In one embodiment, the invention features a method of modulating the expression of a target HDAC gene in a tissue explant comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein one of the siNA strands comprises a sequence complementary to RNA of the target HDAC gene and wherein the sense strand sequence of the siNA comprises a sequence identical or substantially similar to the sequence of the target HDAC RNA; and (b) introducing the siNA molecule into a cell of the tissue explant derived from a particular organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in the tissue explant. In another embodiment, the method further comprises introducing the tissue explant back into the organism the tissue was derived from or into another organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in that organism.

In another embodiment, the invention features a method of modulating the expression of more than one target HDAC gene in a tissue explant comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified, wherein one of the siNA strands comprises a sequence complementary to RNA of the target HDAC genes; and (b) introducing the siNA molecules into a cell of the tissue explant derived from a particular organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC genes in the tissue explant. In another embodiment, the method further comprises introducing the tissue explant back into the organism the tissue was derived from or into another organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC genes in that organism.

In one embodiment, the invention features a method of modulating the expression of a target HDAC gene in a subject or organism comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein one of the siNA strands comprises a sequence complementary to RNA of the target HDAC gene; and (b) introducing the siNA molecule into the subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in the subject or organism. The level of target HDAC protein or RNA can be determined using various methods well-known in the art.

In another embodiment, the invention features a method of modulating the expression of more than one target HDAC gene in a subject or organism comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified, wherein one of the siNA strands comprises a sequence complementary to RNA of the target HDAC genes; and (b) introducing the siNA molecules into the subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC genes in the subject or organism. The level of target HDAC protein or RNA can be determined as is known in the art.

In one embodiment, the invention features a method for modulating the expression of a target HDAC gene within a cell comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the target HDAC gene; and (b) introducing the siNA molecule into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in the cell.

In another embodiment, the invention features a method for modulating the expression of more than one target HDAC gene within a cell comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the target HDAC gene; and (b) contacting the cell in vitro or in vivo with the siNA molecule under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC genes in the cell.

In one embodiment, the invention features a method of modulating the expression of a target HDAC gene in a tissue explant (e.g., a cochlea, skin, heart, liver, spleen, cornea, lung, stomach, kidney, vein, artery, hair, appendage, or limb transplant, or any other organ, tissue or cell as can be transplanted from one organism to another or back to the same organism from which the organ, tissue or cell is derived) comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the target HDAC gene; and (b) contacting a cell of the tissue explant derived from a particular subject or organism with the siNA molecule under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in the tissue explant. In another embodiment, the method further comprises introducing the tissue explant back into the subject or organism the tissue was derived from or into another subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in that subject or organism.

In another embodiment, the invention features a method of modulating the expression of more than one target HDAC gene in a tissue explant (e.g., a cochlear, skin, heart, liver, spleen, cornea, lung, stomach, kidney, vein, artery, hair, appendage, or limb transplant, or any other organ, tissue or cell as can be transplanted from one organism to another or back to the same organism from which the organ, tissue or cell is derived) comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the target HDAC gene; and (b) introducing the siNA molecules into a cell of the tissue explant derived from a particular subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC genes in the tissue explant. In another embodiment, the method further comprises introducing the tissue explant back into the subject or organism the tissue was derived from or into another subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC genes in that subject or organism.

In one embodiment, the invention features a method of modulating the expression of a target HDAC gene in a subject or organism comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the target HDAC gene; and (b) introducing the siNA molecule into the subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in the subject or organism.

In another embodiment, the invention features a method of modulating the expression of more than one target HDAC gene in a subject or organism comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the target HDAC gene; and (b) introducing the siNA molecules into the subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC genes in the subject or organism.

In one embodiment, the invention features a method of modulating the expression of a target HDAC gene in a subject or organism comprising contacting the subject or organism with a siNA molecule of the invention under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC gene in the subject or organism.

In one embodiment, the invention features a method for treating or preventing a disease, disorder, trait or condition related to gene expression in a subject or organism comprising contacting the subject or organism with a siNA molecule of the invention under conditions suitable to modulate the expression of the target HDAC gene in the subject or organism. The reduction of gene expression and thus reduction in the level of the respective protein/RNA relieves, to some extent, the symptoms of the disease, disorder, trait or condition.

In one embodiment, the invention features a method for treating or preventing cancer in a subject or organism comprising contacting the subject or organism with a siNA molecule of the invention under conditions suitable to modulate the expression of the target HDAC gene in the subject or organism whereby the treatment or prevention of cancer can be achieved. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via local administration to relevant tissues or cells, such as cancerous cells and tissues. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via systemic administration (such as via intravenous or subcutaneous administration of siNA) to relevant tissues or cells, such as tissues or cells involved in the maintenance or development of cancer in a subject or organism. The siNA molecule of the invention can be formulated or conjugated as described herein or otherwise known in the art to target HDAC appropriate tisssues or cells in the subject or organism. The siNA molecule can be combined with other therapeutic treatments and modalities as are known in the art for the treatment of or prevention of cancer in a subject or organism.

In one embodiment, the invention features a method for treating or preventing a proliferative disease or condition in a subject or organism comprising contacting the subject or organism with a siNA molecule of the invention under conditions suitable to modulate the expression of the target HDAC gene in the subject or organism whereby the treatment or prevention of the proliferative disease or condition can be achieved. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via local administration to relevant tissues or cells, such as cells and tissues involved in proliferative disease. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via systemic administration (such as via intravenous or subcutaneous administration of siNA) to relevant tissues or cells, such as tissues or cells involved in the maintenance or development of the proliferative disease or condition in a subject or organism. The siNA molecule of the invention can be formulated or conjugated as described herein or otherwise known in the art to target HDAC appropriate tisssues or cells in the subject or organism. The siNA molecule can be combined with other therapeutic treatments and modalities as are known in the art for the treatment of or prevention of proliferative diseases, traits, disorders, or conditions in a subject or organism.

In one embodiment, the invention features a method for treating or preventing an age-related disease, disorder, trait or condition in a subject or organism comprising contacting the subject or organism with a siNA molecule of the invention under conditions suitable to modulate the expression of the target HDAC gene in the subject or organism whereby the treatment or prevention of the age-related disease, disorder, trait or condition can be achieved. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via local administration to relevant tissues or cells, such as cells and tissues involved in the age-related disease, disorder, trait or condition. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via systemic administration (such as via intravenous or subcutaneous administration of siNA) to relevant tissues or cells, such as tissues or cells involved in the maintenance or development of the age-related disease, disorder, trait or condition in a subject or organism. The siNA molecule of the invention can be formulated or conjugated as described herein or otherwise known in the art to target HDAC appropriate tisssues or cells in the subject or organism. The siNA molecule can be combined with other therapeutic treatments and modalities as are known in the art for the treatment of or prevention of age-related diseases, traits, disorders, or conditions in a subject or organism.

In one embodiment, the invention features a method for treating or preventing transplant and/or tissue rejection (allograft rejection) in a subject or organism comprising contacting the subject or organism with a siNA molecule of the invention under conditions suitable to modulate the expression of the target HDAC gene in the subject or organism whereby the treatment or prevention of transplant and/or tissue rejection (allograft rejection) can be achieved. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via local administration to relevant tissues or cells, such as cells and tissues involved in transplant and/or tissue rejection (allograft rejection). In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via systemic administration (such as via intravenous or subcutaneous administration of siNA) to relevant tissues or cells, such as tissues or cells involved in the maintenance or development of transplant and/or tissue rejection (allograft rejection) in a subject or organism. The siNA molecule of the invention can be formulated or conjugated as described herein or otherwise known in the art to target HDAC appropriate tisssues or cells in the subject or organism. The siNA molecule can be combined with other therapeutic treatments and modalities as are known in the art for the treatment of or prevention of transplant and/or tissue rejection (allograft rejection) in a subject or organism.

In one embodiment, the siNA molecule or double stranded nucleic acid molecule of the invention is formulated as a composition described in U.S. Provisional patent application No. 60/678,531 and in related U.S. Provisional patent application No. 60/703,946, both of which are incorporated by reference herein in their entirety.

In one embodiment, the invention features a method for treating or preventing cancer or proliferative disease in a subject, comprising administering to the subject a chemically synthesized double stranded nucleic acid molecule, wherein (a) the double stranded nucleic acid molecule comprises a sense strand and an antisense strand; (b) each strand of the double stranded nucleic acid molecule is 15 to 30 nucleotides in length; (c) at least 15 nucleotides of the sense strand are complementary to the antisense strand; and (d) the antisense strand of the double stranded nucleic acid molecule has complementarity to a HDAC target RNA, wherein the double stranded nucleic acid molecule is administered under conditions suitable for reducing or inhibiting the level of cancer or proliferative disease in the subject compared to a subject not treated with the double stranded nucleic acid molecule.

In one embodiment, the invention features a method for treating or preventing cancer or proliferative disease in a subject, comprising administering to the subject a chemically synthesized double stranded nucleic acid molecule, wherein (a) the double stranded nucleic acid molecule comprises a sense strand and an antisense strand; (b) each strand of the double stranded nucleic acid molecule is 15 to 30 nucleotides in length; (c) at least 15 nucleotides of the sense strand are complementary to the antisense strand; (d) the antisense strand of the double stranded nucleic acid molecule has complementarity to a HDAC target RNA; (e) at least 20% of the internal nucleotides of each strand of the double stranded nucleic acid molecule comprises nucleosides having a chemical modification; and (f) at least two (e.g., 2, 3, 4, 5, or more) of the chemical modifications are different from each other, wherein the double stranded nucleic acid molecule is administered under conditions suitable for reducing or inhibiting the level of cancer or proliferative disease in the subject compared to a subject not treated with the double stranded nucleic acid molecule.

In one embodiment, the invention features a method for treating or preventing cancer or proliferative disease in a subject, comprising administering to the subject a chemically synthesized double stranded nucleic acid molecule, wherein (a) the double stranded nucleic acid molecule comprises a sense strand and an antisense strand; (b) each strand of the double stranded nucleic acid molecule is 15 to 30 nucleotides in length; (c) at least 15 nucleotides of the sense strand are complementary to the antisense strand; (d) the antisense strand of the double stranded nucleic acid molecule has complementarity to a HDAC target RNA; (e) at least 20% of the internal nucleotides of each strand of the double stranded nucleic acid molecule comprises nucleosides having a sugar modification; and (f) at least two (e.g., 2, 3, 4, 5, or more) of the sugar modifications are different from each other, wherein the double stranded nucleic acid molecule is administered under conditions suitable for reducing or inhibiting the level of cancer or proliferative disease in the subject compared to a subject not treated with the double stranded nucleic acid molecule.

In any of the methods of treatment of the invention, the siNA can be administered to the subject as a course of treatment, for example administration at various time intervals, such as once per day over the course of treatment, once every two days over the course of treatment, once every three days over the course of treatment, once every four days over the course of treatment, once every five days over the course of treatment, once every six days over the course of treatment, once per week over the course of treatment, once every other week over the course of treatment, once per month over the course of treatment, etc. In one embodiment, the course of treatment is from about one to about 52 weeks or longer (e.g., indefinitely). In one embodiment, the course of treatment is from about one to about 48 months or longer (e.g., indefinitely).

In any of the methods of treatment of the invention, the siNA can be administered to the subject systemically as described herein or otherwise known in the art, either alone as a monotherapy or in combination with additional therapies as are known in the art. Systemic administration can include, for example, intravenous, subcutaneous, intramuscular, catheterization, nasopharangeal, transdermal, or gastrointestinal administration as is generally known in the art.

In one embodiment, in any of the methods of treatment or prevention of the invention, the siNA can be administered to the subject locally or to local tissues as described herein or otherwise known in the art, either alone as a monotherapy or in combination with additional therapies as are known in the art. Local administration can include, for example, catheterization, implantation, direct injection, dermal/transdermal application, stenting, ear/eye drops, or portal vein administration to relevant tissues, or any other local administration technique, method or procedure, as is generally known in the art.

In another embodiment, the invention features a method of modulating the expression of more than one target HDAC gene in a subject or organism comprising contacting the subject or organism with one or more siNA molecules of the invention under conditions suitable to modulate (e.g., inhibit) the expression of the target HDAC genes in the subject or organism.

The siNA molecules of the invention can be designed to down regulate or inhibit target gene expression through RNAi targeting of a variety of nucleic acid molecules. In one embodiment, the siNA molecules of the invention are used to target various DNA corresponding to a target HDAC gene, for example via heterochromatic silencing. In one embodiment, the siNA molecules of the invention are used to target various RNAs corresponding to a target HDAC gene, for example via RNA target cleavage or translational inhibition. Non-limiting examples of such RNAs include messenger RNA (mRNA), non-coding RNA or regulatory elements, alternate RNA splice variants of target gene(s), post-transcriptionally modified RNA of target gene(s), pre-mRNA of target gene(s), and/or RNA templates. If alternate splicing produces a family of transcripts that are distinguished by usage of appropriate exons, the instant invention can be used to inhibit gene expression through the appropriate exons to specifically inhibit or to distinguish among the functions of HDAC gene family members. For example, a protein that contains an alternatively spliced transmembrane domain can be expressed in both membrane bound and secreted forms. Use of the invention to target the exon containing the transmembrane domain can be used to determine the functional consequences of pharmaceutical targeting of membrane bound as opposed to the secreted form of the protein. Non-limiting examples of applications of the invention relating to targeting these RNA molecules include therapeutic pharmaceutical applications, cosmetic applications, veterinary applications, pharmaceutical discovery applications, molecular diagnostic and gene function applications, and gene mapping, for example using single nucleotide polymorphism mapping with siNA molecules of the invention. Such applications can be implemented using known gene sequences or from partial sequences available from an expressed sequence tag (EST).

In another embodiment, the siNA molecules of the invention are used to target conserved sequences corresponding to a HDAC gene family (e.g., any of class I, class II, and/or class III HDAC genes) or gene families such as HDAC gene families having homologous sequences. As such, siNA molecules targeting multiple HDAC gene or RNA targets can provide increased therapeutic effect. In one embodiment, the invention features the targeting (cleavage or inhibition of expression or function) of more than one target HDAC gene sequence using a single siNA molecule, by targeting the conserved sequences of the targeted HDAC genes.

In addition, siNA can be used to characterize pathways of gene function in a variety of applications. For example, the present invention can be used to inhibit the activity of target gene(s) in a pathway to determine the function of uncharacterized gene(s) in gene function analysis, mRNA function analysis, or translational analysis. The invention can be used to determine potential target gene pathways involved in various diseases and conditions toward pharmaceutical development. The invention can be used to understand pathways of gene expression involved in, for example, the progression and/or maintenance of hearing loss, deafness, tinnitus, movement or balance disorders, and any other diseases, traits, and conditions associated with target gene expression or activity in a subject or organism.

In one embodiment, siNA molecule(s) and/or methods of the invention are used to down regulate the expression of gene(s) that encode RNA referred to by Genbank Accession, for example, target genes encoding RNA sequence(s) referred to herein by Genbank Accession number, for example, Genbank Accession Nos. shown in Table I or U.S. Ser. No. 10/923,536 and U.S. Ser. No. 10/923,536, both incorporated by reference herein.

In one embodiment, the invention features a method comprising: (a) generating a library of siNA constructs having a predetermined complexity; and (b) assaying the siNA constructs of (a) above, under conditions suitable to determine RNAi target sites within the target HDAC RNA sequence. In one embodiment, the siNA molecules of (a) have strands of a fixed length, for example, about 23 nucleotides in length. In another embodiment, the siNA molecules of (a) are of differing length, for example having strands of about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length. In one embodiment, the assay can comprise a reconstituted in vitro siNA assay as described herein. In another embodiment, the assay can comprise a cell culture system in which target HDAC RNA is expressed. In another embodiment, fragments of target HDAC RNA are analyzed for detectable levels of cleavage, for example by gel electrophoresis, northern blot analysis, or RNAse protection assays, to determine the most suitable target site(s) within the target HDAC RNA sequence. The target HDAC RNA sequence can be obtained as is known in the art, for example, by cloning and/or transcription for in vitro systems, and by cellular expression in in vivo systems.

In one embodiment, the invention features a method comprising: (a) generating a randomized library of siNA constructs having a predetermined complexity, such as of 4^N, where N represents the number of base paired nucleotides in each of the siNA construct strands (eg. for a siNA construct having 21 nucleotide sense and antisense strands with 19 base pairs, the complexity would be 4¹⁹); and (b) assaying the siNA constructs of (a) above, under conditions suitable to determine RNAi target sites within the target HDAC RNA sequence. In another embodiment, the siNA molecules of (a) have strands of a fixed length, for example about 23 nucleotides in length. In yet another embodiment, the siNA molecules of (a) are of differing length, for example having strands of about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length. In one embodiment, the assay can comprise a reconstituted in vitro siNA assay as described in Example 6 herein. In another embodiment, the assay can comprise a cell culture system in which target HDAC RNA is expressed. In another embodiment, fragments of target HDAC RNA are analyzed for detectable levels of cleavage, for example, by gel electrophoresis, northern blot analysis, or RNAse protection assays, to determine the most suitable target HDAC site(s) within the target HDAC RNA sequence. The target HDAC RNA sequence can be obtained as is known in the art, for example, by cloning and/or transcription for in vitro systems, and by cellular expression in in vivo systems.

In another embodiment, the invention features a method comprising: (a) analyzing the sequence of a RNA target encoded by a target HDAC gene; (b) synthesizing one or more sets of siNA molecules having sequence complementary to one or more regions of the RNA of (a); and (c) assaying the siNA molecules of (b) under conditions suitable to determine RNAi targets within the target HDAC RNA sequence. In one embodiment, the siNA molecules of (b) have strands of a fixed length, for example about 23 nucleotides in length. In another embodiment, the siNA molecules of (b) are of differing length, for example having strands of about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length. In one embodiment, the assay can comprise a reconstituted in vitro siNA assay as described herein. In another embodiment, the assay can comprise a cell culture system in which target HDAC RNA is expressed. Fragments of target HDAC RNA are analyzed for detectable levels of cleavage, for example by gel electrophoresis, northern blot analysis, or RNAse protection assays, to determine the most suitable target HDAC site(s) within the target HDAC RNA sequence. The target HDAC RNA sequence can be obtained as is known in the art, for example, by cloning and/or transcription for in vitro systems, and by expression in in vivo systems.

By “target site” is meant a sequence within a target RNA (e.g., target HDAC RNA) that is “targeted” for cleavage mediated by a siNA construct which contains sequences within its antisense region that are complementary to the target sequence.

By “detectable level of cleavage” is meant cleavage of target RNA (and formation of cleaved product RNAs) to an extent sufficient to discern cleavage products above the background of RNAs produced by random degradation of the target RNA. Production of cleavage products from 1-5% of the target RNA is sufficient to detect above the background for most methods of detection.

In one embodiment, the invention features a composition comprising a siNA molecule of the invention, which can be chemically-modified, in a pharmaceutically acceptable carrier or diluent. In another embodiment, the invention features a pharmaceutical composition comprising siNA molecules of the invention, which can be chemically-modified, targeting one or more genes in a pharmaceutically acceptable carrier or diluent. In another embodiment, the invention features a method for diagnosing a disease, trait, or condition in a subject comprising administering to the subject a composition of the invention under conditions suitable for the diagnosis of the disease, trait, or condition in the subject. In another embodiment, the invention features a method for treating or preventing a disease, trait, or condition, such as hearing loss, deafness, tinnitus, and/or motion and balance disorders in a subject, comprising administering to the subject a composition of the invention under conditions suitable for the treatment or prevention of the disease, trait, or condition in the subject, alone or in conjunction with one or more other therapeutic compounds.

In another embodiment, the invention features a method for validating a gene target, comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein one of the siNA strands includes a sequence complementary to RNA of a target gene; (b) introducing the siNA molecule into a cell, tissue, subject, or organism under conditions suitable for modulating expression of the target gene in the cell, tissue, subject, or organism; and (c) determining the function of the gene by assaying for any phenotypic change in the cell, tissue, subject, or organism.

In another embodiment, the invention features a method for validating a target comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein one of the siNA strands includes a sequence complementary to RNA of a target gene; (b) introducing the siNA molecule into a biological system under conditions suitable for modulating expression of the target gene in the biological system; and (c) determining the function of the gene by assaying for any phenotypic change in the biological system.

By “biological system” is meant, material, in a purified or unpurified form, from biological sources, including but not limited to human or animal, wherein the system comprises the components required for RNAi activity. The term “biological system” includes, for example, a cell, tissue, subject, or organism, or extract thereof. The term biological system also includes reconstituted RNAi systems that can be used in an in vitro setting.

By “phenotypic change” is meant any detectable change to a cell that occurs in response to contact or treatment with a nucleic acid molecule of the invention (e.g., siNA). Such detectable changes include, but are not limited to, changes in shape, size, proliferation, motility, protein expression or RNA expression or other physical or chemical changes as can be assayed by methods known in the art. The detectable change can also include expression of reporter genes/molecules such as Green Florescent Protein (GFP) or various tags that are used to identify an expressed protein or any other cellular component that can be assayed.

In one embodiment, the invention features a kit containing a siNA molecule of the invention, which can be chemically-modified, that can be used to modulate the expression of a target gene in a biological system, including, for example, in a cell, tissue, subject, or organism. In another embodiment, the invention features a kit containing more than one siNA molecule of the invention, which can be chemically-modified, that can be used to modulate the expression of more than one target gene in a biological system, including, for example, in a cell, tissue, subject, or organism.

In one embodiment, the invention features a cell containing one or more siNA molecules of the invention, which can be chemically-modified. In another embodiment, the cell containing a siNA molecule of the invention is a mammalian cell. In yet another embodiment, the cell containing a siNA molecule of the invention is a human cell.

In one embodiment, the synthesis of a siNA molecule of the invention, which can be chemically-modified, comprises: (a) synthesis of two complementary strands of the siNA molecule; (b) annealing the two complementary strands together under conditions suitable to obtain a double-stranded siNA molecule. In another embodiment, synthesis of the two complementary strands of the siNA molecule is by solid phase oligonucleotide synthesis. In yet another embodiment, synthesis of the two complementary strands of the siNA molecule is by solid phase tandem oligonucleotide synthesis.

In one embodiment, the invention features a method for synthesizing a siNA duplex molecule comprising: (a) synthesizing a first oligonucleotide sequence strand of the siNA molecule, wherein the first oligonucleotide sequence strand comprises a cleavable linker molecule that can be used as a scaffold for the synthesis of the second oligonucleotide sequence strand of the siNA; (b) synthesizing the second oligonucleotide sequence strand of siNA on the scaffold of the first oligonucleotide sequence strand, wherein the second oligonucleotide sequence strand further comprises a chemical moiety than can be used to purify the siNA duplex; (c) cleaving the linker molecule of (a) under conditions suitable for the two siNA oligonucleotide strands to hybridize and form a stable duplex; and (d) purifying the siNA duplex utilizing the chemical moiety of the second oligonucleotide sequence strand. In one embodiment, cleavage of the linker molecule in (c) above takes place during deprotection of the oligonucleotide, for example, under hydrolysis conditions using an alkylamine base such as methylamine. In one embodiment, the method of synthesis comprises solid phase synthesis on a solid support such as controlled pore glass (CPG) or polystyrene, wherein the first sequence of (a) is synthesized on a cleavable linker, such as a succinyl linker, using the solid support as a scaffold. The cleavable linker in (a) used as a scaffold for synthesizing the second strand can comprise similar reactivity as the solid support derivatized linker, such that cleavage of the solid support derivatized linker and the cleavable linker of (a) takes place concomitantly. In another embodiment, the chemical moiety of (b) that can be used to isolate the attached oligonucleotide sequence comprises a trityl group, for example a dimethoxytrityl group, which can be employed in a trityl-on synthesis strategy as described herein. In yet another embodiment, the chemical moiety, such as a dimethoxytrityl group, is removed during purification, for example, using acidic conditions.

In a further embodiment, the method for siNA synthesis is a solution phase synthesis or hybrid phase synthesis wherein both strands of the siNA duplex are synthesized in tandem using a cleavable linker attached to the first sequence which acts a scaffold for synthesis of the second sequence. Cleavage of the linker under conditions suitable for hybridization of the separate siNA sequence strands results in formation of the double-stranded siNA molecule.

In another embodiment, the invention features a method for synthesizing a siNA duplex molecule comprising: (a) synthesizing one oligonucleotide sequence strand of the siNA molecule, wherein the sequence comprises a cleavable linker molecule that can be used as a scaffold for the synthesis of another oligonucleotide sequence; (b) synthesizing a second oligonucleotide sequence having complementarity to the first sequence strand on the scaffold of (a), wherein the second sequence comprises the other strand of the double-stranded siNA molecule and wherein the second sequence further comprises a chemical moiety than can be used to isolate the attached oligonucleotide sequence; (c) purifying the product of (b) utilizing the chemical moiety of the second oligonucleotide sequence strand under conditions suitable for isolating the full-length sequence comprising both siNA oligonucleotide strands connected by the cleavable linker and under conditions suitable for the two siNA oligonucleotide strands to hybridize and form a stable duplex. In one embodiment, cleavage of the linker molecule in (c) above takes place during deprotection of the oligonucleotide, for example, under hydrolysis conditions. In another embodiment, cleavage of the linker molecule in (c) above takes place after deprotection of the oligonucleotide. In another embodiment, the method of synthesis comprises solid phase synthesis on a solid support such as controlled pore glass (CPG) or polystyrene, wherein the first sequence of (a) is synthesized on a cleavable linker, such as a succinyl linker, using the solid support as a scaffold. The cleavable linker in (a) used as a scaffold for synthesizing the second strand can comprise similar reactivity or differing reactivity as the solid support derivatized linker, such that cleavage of the solid support derivatized linker and the cleavable linker of (a) takes place either concomitantly or sequentially. In one embodiment, the chemical moiety of (b) that can be used to isolate the attached oligonucleotide sequence comprises a trityl group, for example a dimethoxytrityl group.

In another embodiment, the invention features a method for making a double-stranded siNA molecule in a single synthetic process comprising: (a) synthesizing an oligonucleotide having a first and a second sequence, wherein the first sequence is complementary to the second sequence, and the first oligonucleotide sequence is linked to the second sequence via a cleavable linker, and wherein a terminal 5′-protecting group, for example, a 5′-O-dimethoxytrityl group (5′-O-DMT) remains on the oligonucleotide having the second sequence; (b) deprotecting the oligonucleotide whereby the deprotection results in the cleavage of the linker joining the two oligonucleotide sequences; and (c) purifying the product of (b) under conditions suitable for isolating the double-stranded siNA molecule, for example using a trityl-on synthesis strategy as described herein.

In another embodiment, the method of synthesis of siNA molecules of the invention comprises the teachings of Scaringe et al., U.S. Pat. Nos. 5,889,136; 6,008,400; and 6,111,086, incorporated by reference herein in their entirety.

In one embodiment, the invention features siNA constructs that mediate RNAi against a target polynucleotide (e.g., HDAC RNA or HDAC DNA target), wherein the siNA construct comprises one or more chemical modifications, for example, one or more chemical modifications having any of Formulae I-VII or any combination thereof that increases the nuclease resistance of the siNA construct.

In another embodiment, the invention features a method for generating siNA molecules with increased nuclease resistance comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having increased nuclease resistance.

In another embodiment, the invention features a method for generating siNA molecules with improved toxicologic profiles (e.g., having attenuated or no immunstimulatory properties) comprising (a) introducing nucleotides having any of Formula I-VII (e.g., siNA motifs referred to in Table IV) or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved toxicologic profiles.

In another embodiment, the invention features a method for generating siNA formulations with improved toxicologic profiles (e.g., having attenuated or no immunstimulatory properties) comprising (a) generating a siNA formulation comprising a siNA molecule of the invention and a delivery vehicle or delivery particle as described herein or as otherwise known in the art, and (b) assaying the siNA formualtion of step (a) under conditions suitable for isolating siNA formulations having improved toxicologic profiles.

In another embodiment, the invention features a method for generating siNA molecules that do not stimulate an interferon response (e.g., no interferon response or attenuated interferon response) in a cell, subject, or organism, comprising (a) introducing nucleotides having any of Formula I-VII (e.g., siNA motifs referred to in Table IV) or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules that do not stimulate an interferon response.

In another embodiment, the invention features a method for generating siNA formulations that do not stimulate an interferon response (e.g., no interferon response or attenuated interferon response) in a cell, subject, or organism, comprising (a) generating a siNA formulation comprising a siNA molecule of the invention and a delivery vehicle or delivery particle as described herein or as otherwise known in the art, and (b) assaying the siNA formualtion of step (a) under conditions suitable for isolating siNA formulations that do not stimulate an interferon response. In one embodiment, the interferon comprises interferon alpha.

In another embodiment, the invention features a method for generating siNA molecules that do not stimulate an inflammatory or proinflammatory cytokine response (e.g., no cytokine response or attenuated cytokine response) in a cell, subject, or organism, comprising (a) introducing nucleotides having any of Formula I-VII (e.g., siNA motifs referred to in Table IV) or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules that do not stimulate a cytokine response. In one embodiment, the cytokine comprises an interleukin such as interleukin-6 (IL-6) and/or tumor necrosis alpha (TNF-α).

In another embodiment, the invention features a method for generating siNA formulations that do not stimulate an inflammatory or proinflammatory cytokine response (e.g., no cytokine response or attenuated cytokine response) in a cell, subject, or organism, comprising (a) generating a siNA formulation comprising a siNA molecule of the invention and a delivery vehicle or delivery particle as described herein or as otherwise known in the art, and (b) assaying the siNA formualtion of step (a) under conditions suitable for isolating siNA formulations that do not stimulate a cytokine response. In one embodiment, the cytokine comprises an interleukin such as interleukin-6 (IL-6) and/or tumor necrosis alpha (TNF-α).

In another embodiment, the invention features a method for generating siNA molecules that do not stimulate Toll-like Receptor (TLR) response (e.g., no TLR response or attenuated TLR response) in a cell, subject, or organism, comprising (a) introducing nucleotides having any of Formula I-VII (e.g., siNA motifs referred to in Table IV) or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules that do not stimulate a TLR response. In one embodiment, the TLR comprises TLR3, TLR7, TLR8 and/or TLR9.

In another embodiment, the invention features a method for generating siNA formulations that do not stimulate a Toll-like Receptor (TLR) response (e.g., no TLR response or attenuated TLR response) in a cell, subject, or organism, comprising (a) generating a siNA formulation comprising a siNA molecule of the invention and a delivery vehicle or delivery particle as described herein or as otherwise known in the art, and (b) assaying the siNA formualtion of step (a) under conditions suitable for isolating siNA formulations that do not stimulate a TLR response. In one embodiment, the TLR comprises TLR3, TLR7, TLR8 and/or TLR9.

In one embodiment, the invention features a chemically synthesized double stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a target RNA via RNA interference (RNAi), wherein: (a) each strand of said siNA molecule is about 18 to about 38 nucleotides in length; (b) one strand of said siNA molecule comprises nucleotide sequence having sufficient complementarity to said target RNA for the siNA molecule to direct cleavage of the target RNA via RNA interference; and (c) wherein the nucleotide positions within said siNA molecule are chemically modified to reduce the immunostimulatory properties of the siNA molecule to a level below that of a corresponding unmodified siRNA molecule. Such siNA molecules are said to have an improved toxicologic profile compared to an unmodified or minimally modified siNA.

By “improved toxicologic profile”, is meant that the chemically modified or formulated siNA construct exhibits decreased toxicity in a cell, subject, or organism compared to an unmodified or unformulated siNA, or siNA molecule having fewer modifications or modifications that are less effective in imparting improved toxicology. In a non-limiting example, siNA molecules and formulations with improved toxicologic profiles are associated with reduced immunostimulatory properties, such as a reduced, decreased or attenuated immunostimulatory response in a cell, subject, or organism compared to an unmodified or unformulated siNA, or siNA molecule having fewer modifications or modifications that are less effective in imparting improved toxicology. Such an improved toxicologic profile is characterized by abrogated or reduced immunostimulation, such as reduction or abrogation of induction of interferons (e.g., interferon alpha), inflammatory cytokines (e.g., interleukins such as IL-6, and/or TNF-alpha), and/or toll like receptors (e.g., TLR-3, TLR-7, TLR-8, and/or TLR-9). In one embodiment, a siNA molecule or formulation with an improved toxicological profile comprises no ribonucleotides. In one embodiment, a siNA molecule or formulation with an improved toxicological profile comprises less than 5 ribonucleotides (e.g., 1, 2, 3, or 4 ribonucleotides). In one embodiment, a siNA molecule or formulation with an improved toxicological profile comprises Stab 7, Stab 8, Stab 11, Stab 12, Stab 13, Stab 16, Stab 17, Stab 18, Stab 19, Stab 20, Stab 23, Stab 24, Stab 25, Stab 26, Stab 27, Stab 28, Stab 29, Stab 30, Stab 31, Stab 32, Stab 33, Stab 34 or any combination thereof (see Table IV). Herein, numeric Stab chemistries include both 2′-fluoro and 2′-OCF3 versions of the chemistries shown in Table IV. For example, “Stab 7/8” refers to both Stab 7/8 and Stab 7F/8F etc. In one embodiment, a siNA molecule or formulation with an improved toxicological profile comprises a siNA molecule of the invention and a formulation as described in United States Patent Application Publication No. 20030077829, incorporated by reference herein in its entirety including the drawings.

In one embodiment, the level of immunostimulatory response associated with a given siNA molecule can be measured as is described herein or as is otherwise known in the art, for example by determining the level of PKR/interferon response, proliferation, B-cell activation, and/or cytokine production in assays to quantitate the immunostimulatory response of particular siNA molecules (see, for example, Leifer et al., 2003, J Immunother. 26, 313-9; and U.S. Pat. No. 5,968,909, incorporated in its entirety by reference). In one embodiment, the reduced immunostimulatory response is between about 10% and about 100% compared to an unmodified or minimally modified siRNA molecule, e.g., about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduced immunostimulatory response. In one embodiment, the immunostimulatory response associated with a siNA molecule can be modulated by the degree of chemical modification. For example, a siNA molecule having between about 10% and about 100%, e.g., about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% of the nucleotide positions in the siNA molecule modified can be selected to have a corresponding degree of immunostimulatory properties as described herein.

In one embodiment, the degree of reduced immunostimulatory response is selected for optimized RNAi activity. For example, retaining a certain degree of immunostimulation can be preferred to treat viral infection, where less than 100% reduction in immunostimulation may be preferred for maximal antiviral activity (e.g., about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% reduction in immunostimulation) whereas the inhibition of expression of an endogenous gene target may be preferred with siNA molecules that posess minimal immunostimulatory properties to prevent non-specific toxicity or off target effects (e.g., about 90% to about 100% reduction in immunostimulation).

In one embodiment, the invention features a chemically synthesized double stranded siNA molecule that directs cleavage of a target HDAC RNA via RNA interference (RNAi), wherein (a) each strand of said siNA molecule is about 18 to about 38 nucleotides in length; (b) one strand of said siNA molecule comprises nucleotide sequence having sufficient complementarity to said target RNA for the siNA molecule to direct cleavage of the target HDAC RNA via RNA interference; and (c) wherein one or more nucleotides of said siNA molecule are chemically modified to reduce the immunostimulatory properties of the siNA molecule to a level below that of a corresponding unmodified siNA molecule. In one embodiment, each starnd comprises at least about 18 nucleotides that are complementary to the nucleotides of the other strand.

In another embodiment, the siNA molecule comprising modified nucleotides to reduce the immunostimulatory properties of the siNA molecule comprises an antisense region having nucleotide sequence that is complemetary to a nucleotide sequence of a target gene or a protion thereof and further comprises a sense region, wherein said sense region comprises a nucleotide sequence substantially similar to the nucleotide sequence of said target gene or protion thereof. In one embodiment thereof, the antisense region and the sense region comprise about 18 to about 38 nucleotides, wherein said antisense region comprises at least about 18 nucleotides that are complementary to nucleotides of the sense region. In one embodiment thereof, the pyrimidine nucleotides in the sense region are 2′-O-methyl pyrimidine nucleotides. In another embodiment thereof, the purine nucleotides in the sense region are 2′-deoxy purine nucleotides. In yet another embodiment thereof, the pyrimidine nucleotides present in the sense region are 2′-deoxy-2′-fluoro pyrimidine nucleotides. In another embodiment thereof, the pyrimidine nucleotides of said antisense region are 2′-deoxy-2′-fluoro pyrimidine nucleotides. In yet another embodiment thereof, the purine nucleotides of said antisense region are 2′-O-methyl purine nucleotides. In still another embodiment thereof, the purine nucleotides present in said antisense region comprise 2′-deoxypurine nucleotides. In another embodiment, the antisense region comprises a phosphorothioate internucleotide linkage at the 3′ end of said antisense region. In another embodiment, the antisense region comprises a glyceryl modification at a 3′ end of said antisense region.

In other embodiments, the siNA molecule comprisisng modified nucleotides to reduce the immunostimulatory properties of the siNA molecule can comprise any of the structural features of siNA molecules described herein. In other embodiments, the siNA molecule comprising modified nucleotides to reduce the immunostimulatory properties of the siNA molecule can comprise any of the chemical modifications of siNA molecules described herein.

In one embodiment, the invention features a method for generating a chemically synthesized double stranded siNA molecule having chemically modified nucleotides to reduce the immunostimulatory properties of the siNA molecule, comprising (a) introducing one or more modified nucleotides in the siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating an siNA molecule having reduced immunostimulatory properties compared to a corresponding siNA molecule having unmodified nucleotides. Each strand of the siNA molecule is about 18 to about 38 nucleotides in length. One strand of the siNA molecule comprises nucleotide sequence having sufficient complementarity to the target RNA for the siNA molecule to direct cleavage of the target HDAC RNA via RNA interference. In one embodiment, the reduced immunostimulatory properties comprise an abrogated or reduced induction of inflammatory or proinflammatory cytokines, such as interleukin-6 (IL-6) or tumor necrosis alpha (TNF-α), in response to the siNA being introduced in a cell, tissue, or organism. In another embodiment, the reduced immunostimulatory properties comprise an abrogated or reduced induction of Toll Like Receptors (TLRs), such as TLR3, TLR7, TLR8 or TLR9, in response to the siNA being introduced in a cell, tissue, or organism. In another embodiment, the reduced immunostimulatory properties comprise an abrogated or reduced induction of interferons, such as interferon alpha, in response to the siNA being introduced in a cell, tissue, or organism.

In another embodiment, the invention features a method for generating siNA molecules with increased binding affinity between the sense and antisense strands of the siNA molecule comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having increased binding affinity between the sense and antisense strands of the siNA molecule.

In one embodiment, the invention features siNA constructs that mediate RNAi against a target HDAC polynucleotide, wherein the siNA construct comprises one or more chemical modifications described herein that modulates the binding affinity between the antisense strand of the siNA construct and a complementary target RNA sequence within a cell.

In one embodiment, the invention features siNA constructs that mediate RNAi against a target HDAC polynucleotide, wherein the siNA construct comprises one or more chemical modifications described herein that modulates the binding affinity between the antisense strand of the siNA construct and a complementary target HDAC DNA sequence within a cell.

In another embodiment, the invention features a method for generating siNA molecules with increased binding affinity between the antisense strand of the siNA molecule and a complementary target HDAC RNA sequence comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having increased binding affinity between the antisense strand of the siNA molecule and a complementary target HDAC RNA sequence.

In another embodiment, the invention features a method for generating siNA molecules with increased binding affinity between the antisense strand of the siNA molecule and a complementary target HDAC DNA sequence comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having increased binding affinity between the antisense strand of the siNA molecule and a complementary target HDAC DNA sequence.

In one embodiment, the invention features siNA constructs that mediate RNAi against a target HDAC polynucleotide, wherein the siNA construct comprises one or more chemical modifications described herein that modulate the polymerase activity of a cellular polymerase capable of generating additional endogenous siNA molecules having sequence homology to the chemically-modified siNA construct.

In another embodiment, the invention features a method for generating siNA molecules capable of mediating increased polymerase activity of a cellular polymerase capable of generating additional endogenous siNA molecules having sequence homology to a chemically-modified siNA molecule comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules capable of mediating increased polymerase activity of a cellular polymerase capable of generating additional endogenous siNA molecules having sequence homology to the chemically-modified siNA molecule.

In one embodiment, the invention features chemically-modified siNA constructs that mediate RNAi against a target HDAC polynucleotide in a cell, wherein the chemical modifications do not significantly effect the interaction of siNA with a target HDAC RNA molecule, DNA molecule and/or proteins or other factors that are essential for RNAi in a manner that would decrease the efficacy of RNAi mediated by such siNA constructs.

In another embodiment, the invention features a method for generating siNA molecules with improved RNAi specificity against polynucleotide HDAC targets comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved RNAi specificity. In one embodiment, improved specificity comprises having reduced off target effects compared to an unmodified siNA molecule. For example, introduction of terminal cap moieties at the 3′-end, 5′-end, or both 3′ and 5′-ends of the sense strand or region of a siNA molecule of the invention can direct the siNA to have improved specificity by preventing the sense strand or sense region from acting as a template for RNAi activity against a corresponding target having complementarity to the sense strand or sense region.

In another embodiment, the invention features a method for generating siNA molecules with improved RNAi activity against a target HDAC polynucleotide comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved RNAi activity.

In yet another embodiment, the invention features a method for generating siNA molecules with improved RNAi activity against a target HDAC RNA comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved RNAi activity against the target HDAC RNA.

In yet another embodiment, the invention features a method for generating siNA molecules with improved RNAi activity against a target HDAC DNA comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved RNAi activity against the target HDAC DNA.

In another embodiment, the invention features a method for generating siNA molecules against a target HDAC polynucleotide with improved cellular uptake comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved cellular uptake.

In one embodiment, the invention features siNA constructs that mediate RNAi against a target HDAC polynucleotide, wherein the siNA construct comprises one or more chemical modifications described herein that increases the bioavailability of the siNA construct, for example, by attaching polymeric conjugates such as polyethyleneglycol or equivalent conjugates that improve the pharmacokinetics of the siNA construct, or by attaching conjugates that target specific tissue types or cell types in vivo. Non-limiting examples of such conjugates are described in Vargeese et al., U.S. Ser. No. 10/201,394 incorporated by reference herein.

In one embodiment, the invention features a method for generating siNA molecules of the invention with improved bioavailability comprising (a) introducing a conjugate into the structure of a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved bioavailability. Such conjugates can include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; cholesterol derivatives, polyamines, such as spermine or spermidine; and others.

In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that comprises a first nucleotide sequence complementary to a target HDAC RNA sequence or a portion thereof, and a second sequence having complementarity to said first sequence, wherein said second sequence is chemically modified in a manner that it can no longer act as a guide sequence for efficiently mediating RNA interference and/or be recognized by cellular proteins that facilitate RNAi. In one embodiment, the first nucleotide sequence of the siNA is chemically modified as described herein. In one embodiment, the first nucleotide sequence of the siNA is not modified (e.g., is all RNA).

In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that comprises a first nucleotide sequence complementary to a target HDAC RNA sequence or a portion thereof, and a second sequence having complementarity to said first sequence, wherein the second sequence is designed or modified in a manner that prevents its entry into the RNAi pathway as a guide sequence or as a sequence that is complementary to a target nucleic acid (e.g., RNA) sequence. In one embodiment, the first nucleotide sequence of the siNA is chemically modified as described herein. In one embodiment, the first nucleotide sequence of the siNA is not modified (e.g., is all RNA). Such design or modifications are expected to enhance the activity of siNA and/or improve the specificity of siNA molecules of the invention. These modifications are also expected to minimize any off-target effects and/or associated toxicity.

In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that comprises a first nucleotide sequence complementary to a target HDAC RNA sequence or a portion thereof, and a second sequence having complementarity to said first sequence, wherein said second sequence comprises a terminal cap moiety at the 5′-end of said second sequence. In one embodiment, the terminal cap moiety comprises an inverted abasic, inverted deoxy abasic, inverted nucleotide moiety, a group shown in FIG. 10, an alkyl or cycloalkyl group, a heterocycle, or any other group that prevents RNAi activity in which the second sequence serves as a guide sequence or template for RNAi.

In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that comprises a first nucleotide sequence complementary to a target HDAC RNA sequence or a portion thereof, and a second sequence having complementarity to said first sequence, wherein said second sequence comprises a terminal cap moiety at the 5′-end and 3′-end of said second sequence. In one embodiment, each terminal cap moiety individually comprises an inverted abasic, inverted deoxy abasic, inverted nucleotide moiety, a group shown in FIG. 10, an alkyl or cycloalkyl group, a heterocycle, or any other group that prevents RNAi activity in which the second sequence serves as a guide sequence or template for RNAi.

In one embodiment, the invention features a method for generating siNA molecules of the invention with improved specificity for down regulating or inhibiting the expression of a target nucleic acid (e.g., a HDAC DNA or HDAC RNA such as a HDAC gene or its corresponding RNA), comprising (a) introducing one or more chemical modifications into the structure of a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved specificity. In another embodiment, the chemical modification used to improve specificity comprises terminal cap modifications at the 5′-end, 3′-end, or both 5′ and 3′-ends of the siNA molecule. The terminal cap modifications can comprise, for example, structures shown in FIG. 10 (e.g. inverted deoxyabasic moieties) or any other chemical modification that renders a portion of the siNA molecule (e.g. the sense strand) incapable of mediating RNA interference against an off target nucleic acid sequence. In a non-limiting example, a siNA molecule is designed such that only the antisense sequence of the siNA molecule can serve as a guide sequence for RISC mediated degradation of a corresponding target RNA sequence. This can be accomplished by rendering the sense sequence of the siNA inactive by introducing chemical modifications to the sense strand that preclude recognition of the sense strand as a guide sequence by RNAi machinery. In one embodiment, such chemical modifications comprise any chemical group at the 5′-end of the sense strand of the siNA, or any other group that serves to render the sense strand inactive as a guide sequence for mediating RNA interference. These modifications, for example, can result in a molecule where the 5′-end of the sense strand no longer has a free 5′-hydroxyl (5′-OH) or a free 5′-phosphate group (e.g., phosphate, diphosphate, triphosphate, cyclic phosphate etc.). Non-limiting examples of such siNA constructs are described herein, such as “Stab 9/10”, “Stab 7/8”, “Stab 7/19”, “Stab 17/22”, “Stab 23/24”, “Stab 24/25”, and “Stab 24/26” (e.g., any siNA having Stab 7, 9, 17, 23, or 24 sense strands) chemistries and variants thereof (see Table IV) wherein the 5′-end and 3′-end of the sense strand of the siNA do not comprise a hydroxyl group or phosphate group. Herein, numeric Stab chemistries include both 2′-fluoro and 2′-OCF3 versions of the chemistries shown in Table IV. For example, “Stab 7/8” refers to both Stab 7/8 and Stab 7F/8F etc.

In one embodiment, the invention features a method for screening siNA molecules that are active in mediating RNA interference against a target HDAC nucleic acid sequence comprising (a) generating a plurality of unmodified siNA molecules, (b) screening the siNA molecules of step (a) under conditions suitable for isolating siNA molecules that are active in mediating RNA interference against the target HDAC nucleic acid sequence, and (c) introducing chemical modifications (e.g. chemical modifications as described herein or as otherwise known in the art) into the active siNA molecules of (b). In one embodiment, the method further comprises re-screening the chemically modified siNA molecules of step (c) under conditions suitable for isolating chemically modified siNA molecules that are active in mediating RNA interference against the target HDAC nucleic acid sequence.

In one embodiment, the invention features a method for screening chemically modified siNA molecules that are active in mediating RNA interference against a target HDAC nucleic acid sequence comprising (a) generating a plurality of chemically modified siNA molecules (e.g. siNA molecules as described herein or as otherwise known in the art), and (b) screening the siNA molecules of step (a) under conditions suitable for isolating chemically modified siNA molecules that are active in mediating RNA interference against the target HDAC nucleic acid sequence.

The term “ligand” refers to any compound or molecule, such as a drug, peptide, hormone, or neurotransmitter, that is capable of interacting with another compound, such as a receptor, either directly or indirectly. The receptor that interacts with a ligand can be present on the surface of a cell or can alternately be an intercellular receptor. Interaction of the ligand with the receptor can result in a biochemical reaction, or can simply be a physical interaction or association.

In another embodiment, the invention features a method for generating siNA molecules of the invention with improved bioavailability comprising (a) introducing an excipient formulation to a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved bioavailability. Such excipients include polymers such as cyclodextrins, lipids, cationic lipids, polyamines, phospholipids, nanoparticles, receptors, ligands, and others.

In another embodiment, the invention features a method for generating siNA molecules of the invention with improved bioavailability comprising (a) introducing nucleotides having any of Formulae I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved bioavailability.

In another embodiment, polyethylene glycol (PEG) can be covalently attached to siNA compounds of the present invention. The attached PEG can be any molecular weight, preferably from about 100 to about 50,000 daltons (Da).

The present invention can be used alone or as a component of a kit having at least one of the reagents necessary to carry out the in vitro or in vivo introduction of RNA to test samples and/or subjects. For example, preferred components of the kit include a siNA molecule of the invention and a vehicle that promotes introduction of the siNA into cells of interest as described herein (e.g., using lipids and other methods of transfection known in the art, see for example Beigelman et al, U.S. Pat. No. 6,395,713). The kit can be used for target validation, such as in determining gene function and/or activity, or in drug optimization, and in drug discovery (see for example Usman et al., U.S. Ser. No. 60/402,996). Such a kit can also include instructions to allow a user of the kit to practice the invention.

The term “short interfering nucleic acid”, “siNA”, “short interfering RNA”, “siRNA”, “short interfering nucleic acid molecule”, “short interfering oligonucleotide molecule”, or “chemically-modified short interfering nucleic acid molecule” as used herein refers to any nucleic acid molecule capable of inhibiting or down regulating gene expression or viral replication, for example by mediating RNA interference “RNAi” or gene silencing in a sequence-specific manner; see for example Zamore et al., 2000, Cell, 101, 25-33; Bass, 2001, Nature, 411, 428-429; Elbashir et al., 2001, Nature, 411, 494-498; and Kreutzer et al., International PCT Publication No. WO 00/44895; Zernicka-Goetz et al., International PCT Publication No. WO 01/36646; Fire, International PCT Publication No. WO 99/32619; Plaetinck et al., International PCT Publication No. WO 00/01846; Mello and Fire, International PCT Publication No. WO 01/29058; Deschamps-Depaillette, International PCT Publication No. WO 99/07409; and Li et al., International PCT Publication No. WO 00/44914; Allshire, 2002, Science, 297, 1818-1819; Volpe et al., 2002, Science, 297, 1833-1837; Jenuwein, 2002, Science, 297, 2215-2218; and Hall et al., 2002, Science, 297, 2232-2237; Hutvagner and Zamore, 2002, Science, 297, 2056-60; McManus et al., 2002, RNA, 8, 842-850; Reinhart et al., 2002, Gene & Dev., 16, 1616-1626; and Reinhart & Bartel, 2002, Science, 297, 1831). Non limiting examples of siNA molecules of the invention are shown in FIGS. 4-6, and Tables II-III herein. For example the siNA can be a double-stranded polynucleotide molecule comprising self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense region having nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. The siNA can be assembled from two separate oligonucleotides, where one strand is the sense strand and the other is the antisense strand, wherein the antisense and sense strands are self-complementary (i.e., each strand comprises nucleotide sequence that is complementary to nucleotide sequence in the other strand; such as where the antisense strand and sense strand form a duplex or double stranded structure, for example wherein the double stranded region is about 15 to about 30, e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 base pairs; the antisense strand comprises nucleotide sequence that is complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense strand comprises nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof (e.g., about 15 to about 25 or more nucleotides of the siNA molecule are complementary to the target nucleic acid or a portion thereof). Alternatively, the siNA is assembled from a single oligonucleotide, where the self-complementary sense and antisense regions of the siNA are linked by means of a nucleic acid based or non-nucleic acid-based linker(s). The siNA can be a polynucleotide with a duplex, asymmetric duplex, hairpin or asymmetric hairpin secondary structure, having self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to nucleotide sequence in a separate target nucleic acid molecule or a portion thereof and the sense region having nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. The siNA can be a circular single-stranded polynucleotide having two or more loop structures and a stem comprising self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense region having nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof, and wherein the circular polynucleotide can be processed either in vivo or in vitro to generate an active siNA molecule capable of mediating RNAi. The siNA can also comprise a single stranded polynucleotide having nucleotide sequence complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof (for example, where such siNA molecule does not require the presence within the siNA molecule of nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof), wherein the single stranded polynucleotide can further comprise a terminal phosphate group, such as a 5′-phosphate (see for example Martinez et al., 2002, Cell., 110, 563-574 and Schwarz et al., 2002, Molecular Cell, 10, 537-568), or 5′,3′-diphosphate. In certain embodiments, the siNA molecule of the invention comprises separate sense and antisense sequences or regions, wherein the sense and antisense regions are covalently linked by nucleotide or non-nucleotide linkers molecules as is known in the art, or are alternately non-covalently linked by ionic interactions, hydrogen bonding, van der waals interactions, hydrophobic interactions, and/or stacking interactions. In certain embodiments, the siNA molecules of the invention comprise nucleotide sequence that is complementary to nucleotide sequence of a target gene. In another embodiment, the siNA molecule of the invention interacts with nucleotide sequence of a target gene in a manner that causes inhibition of expression of the target gene. As used herein, siNA molecules need not be limited to those molecules containing only RNA, but further encompasses chemically-modified nucleotides and non-nucleotides. In certain embodiments, the short interfering nucleic acid molecules of the invention lack 2′-hydroxy (2′-OH) containing nucleotides. Applicant describes in certain embodiments short interfering nucleic acids that do not require the presence of nucleotides having a 2′-hydroxy group for mediating RNAi and as such, short interfering nucleic acid molecules of the invention optionally do not include any ribonucleotides (e.g., nucleotides having a 2′-OH group). Such siNA molecules that do not require the presence of ribonucleotides within the siNA molecule to support RNAi can however have an attached linker or linkers or other attached or associated groups, moieties, or chains containing one or more nucleotides with 2′-OH groups. Optionally, siNA molecules can comprise ribonucleotides at about 5, 10, 20, 30, 40, or 50% of the nucleotide positions. The modified short interfering nucleic acid molecules of the invention can also be referred to as short interfering modified oligonucleotides “siMON.” As used herein, the term siNA is meant to be equivalent to other terms used to describe nucleic acid molecules that are capable of mediating sequence specific RNAi, for example short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), short hairpin RNA (shRNA), short interfering oligonucleotide, short interfering nucleic acid, short interfering modified oligonucleotide, chemically-modified siRNA, post-transcriptional gene silencing RNA (ptgsRNA), and others. In addition, as used herein, the term RNAi is meant to be equivalent to other terms used to describe sequence specific RNA interference, such as post transcriptional gene silencing, translational inhibition, or epigenetics. For example, siNA molecules of the invention can be used to epigenetically silence genes at both the post-transcriptional level or the pre-transcriptional level. In a non-limiting example, epigenetic modulation of gene expression by siNA molecules of the invention can result from siNA mediated modification of chromatin structure or methylation pattern to alter gene expression (see, for example, Verdel et al., 2004, Science, 303, 672-676; Pal-Bhadra et al., 2004, Science, 303, 669-672; Allshire, 2002, Science, 297, 1818-1819; Volpe et al., 2002, Science, 297, 1833-1837; Jenuwein, 2002, Science, 297, 2215-2218; and Hall et al., 2002, Science, 297, 2232-2237). In another non-limiting example, modulation of gene expression by siNA molecules of the invention can result from siNA mediated cleavage of RNA (either coding or non-coding RNA) via RISC, or alternately, translational inhibition as is known in the art.

In one embodiment, a siNA molecule of the invention is a duplex forming oligonucleotide “DFO”, (see for example FIGS. 14-15 and Vaish et al., U.S. Ser. No. 10/727,780 filed Dec. 3, 2003 and International PCT Application No. US04/16390, filed May 24, 2004).

In one embodiment, a siNA molecule of the invention is a multifunctional siNA, (see for example FIGS. 16-21 and Jadhav et al., U.S. Ser. No. 60/543,480 filed Feb. 10, 2004 and International PCT Application No. US04/16390, filed May 24, 2004). In one embodiment, the multifunctional siNA of the invention can comprise sequence targeting, for example, two or more regions of target RNA (see for example target sequences in Tables II and III).

By “asymmetric hairpin” as used herein is meant a linear siNA molecule comprising an antisense region, a loop portion that can comprise nucleotides or non-nucleotides, and a sense region that comprises fewer nucleotides than the antisense region to the extent that the sense region has enough complementary nucleotides to base pair with the antisense region and form a duplex with loop. For example, an asymmetric hairpin siNA molecule of the invention can comprise an antisense region having length sufficient to mediate RNAi in a cell or in vitro system (e.g. about 15 to about 30, or about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides) and a loop region comprising about 4 to about 12 (e.g., about 4, 5, 6, 7, 8, 9, 10, 11, or 12) nucleotides, and a sense region having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) nucleotides that are complementary to the antisense region. The asymmetric hairpin siNA molecule can also comprise a 5′-terminal phosphate group that can be chemically modified. The loop portion of the asymmetric hairpin siNA molecule can comprise nucleotides, non-nucleotides, linker molecules, or conjugate molecules as described herein.

By “asymmetric duplex” as used herein is meant a siNA molecule having two separate strands comprising a sense region and an antisense region, wherein the sense region comprises fewer nucleotides than the antisense region to the extent that the sense region has enough complementary nucleotides to base pair with the antisense region and form a duplex. For example, an asymmetric duplex siNA molecule of the invention can comprise an antisense region having length sufficient to mediate RNAi in a cell or in vitro system (e.g., about 15 to about 30, or about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides) and a sense region having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) nucleotides that are complementary to the antisense region.

By “modulate” is meant that the expression of the gene, or level of a RNA molecule or equivalent RNA molecules encoding one or more proteins or protein subunits, or activity of one or more proteins or protein subunits is up regulated or down regulated, such that expression, level, or activity is greater than or less than that observed in the absence of the modulator. For example, the term “modulate” can mean “inhibit,” but the use of the word “modulate” is not limited to this definition.

By “inhibit”, “down-regulate”, or “reduce”, it is meant that the expression of the gene, or level of RNA molecules or equivalent RNA molecules encoding one or more proteins or protein subunits, or activity of one or more proteins or protein subunits, is reduced below that observed in the absence of the nucleic acid molecules (e.g., siNA) of the invention. In one embodiment, inhibition, down-regulation or reduction with an siNA molecule is below that level observed in the presence of an inactive or attenuated molecule. In another embodiment, inhibition, down-regulation, or reduction with siNA molecules is below that level observed in the presence of, for example, an siNA molecule with scrambled sequence or with mismatches. In another embodiment, inhibition, down-regulation, or reduction of gene expression with a nucleic acid molecule of the instant invention is greater in the presence of the nucleic acid molecule than in its absence. In one embodiment, inhibition, down regulation, or reduction of gene expression is associated with post transcriptional silencing, such as RNAi mediated cleavage of a target nucleic acid molecule (e.g. RNA) or inhibition of translation. In one embodiment, inhibition, down regulation, or reduction of gene expression is associated with pretranscriptional silencing, such as by alterations in DNA methylation patterns and DNA chromatin structure.

By “gene”, or “target gene” or “target DNA”, is meant a nucleic acid that encodes an RNA, for example, nucleic acid sequences including, but not limited to, structural genes encoding a polypeptide. A gene or target gene can also encode a functional RNA (fRNA) or non-coding RNA (ncRNA), such as small temporal RNA (stRNA), micro RNA (miRNA), small nuclear RNA (snRNA), short interfering RNA (siRNA), small nucleolar RNA (snRNA), ribosomal RNA (rRNA), transfer RNA (tRNA) and precursor RNAs thereof. Such non-coding RNAs can serve as target nucleic acid molecules for siNA mediated RNA interference in modulating the activity of fRNA or ncRNA involved in functional or regulatory cellular processes. Abberant fRNA or ncRNA activity leading to disease can therefore be modulated by siNA molecules of the invention. siNA molecules targeting fRNA and ncRNA can also be used to manipulate or alter the genotype or phenotype of a subject, organism or cell, by intervening in cellular processes such as genetic imprinting, transcription, translation, or nucleic acid processing (e.g., transamination, methylation etc.). The target gene can be a gene derived from a cell, an endogenous gene, a transgene, or exogenous genes such as genes of a pathogen, for example a virus, which is present in the cell after infection thereof. The cell containing the target gene can be derived from or contained in any organism, for example a plant, animal, protozoan, virus, bacterium, or fungus. Non-limiting examples of plants include monocots, dicots, or gymnosperms. Non-limiting examples of animals include vertebrates or invertebrates. Non-limiting examples of fungi include molds or yeasts. For a review, see for example Snyder and Gerstein, 2003, Science, 300, 258-260.

By “non-canonical base pair” is meant any non-Watson Crick base pair, such as mismatches and/or wobble base pairs, including flipped mismatches, single hydrogen bond mismatches, trans-type mismatches, triple base interactions, and quadruple base interactions. Non-limiting examples of such non-canonical base pairs include, but are not limited to, AC reverse Hoogsteen, AC wobble, AU reverse Hoogsteen, GU wobble, AA N7 amino, CC 2-carbonyl-amino(H1)-N-3-amino(H2), GA sheared, UC 4-carbonyl-amino, UU imino-carbonyl, AC reverse wobble, AU Hoogsteen, AU reverse Watson Crick, CG reverse Watson Crick, GC N3-amino-amino N3, AA N1-amino symmetric, AA N7-amino symmetric, GA N7-N1 amino-carbonyl, GA+ carbonyl-amino N7-N1, GG N1-carbonyl symmetric, GG N3-amino symmetric, CC carbonyl-amino symmetric, CC N3-amino symmetric, UU 2-carbonyl-imino symmetric, UU 4-carbonyl-imino symmetric, AA amino-N3, AA N1-amino, AC amino 2-carbonyl, AC N3-amino, AC N7-amino, AU amino-4-carbonyl, AU N1-imino, AU N3-imino, AU N7-imino, CC carbonyl-amino, GA amino-N1, GA amino-N7, GA carbonyl-amino, GA N3-amino, GC amino-N3, GC carbonyl-amino, GC N3-amino, GC N7-amino, GG amino-N7, GG carbonyl-imino, GG N7-amino, GU amino-2-carbonyl, GU carbonyl-imino, GU imino-2-carbonyl, GU N7-imino, psiU imino-2-carbonyl, UC 4-carbonyl-amino, UC imino-carbonyl, UU imino-4-carbonyl, AC C2-H-N3, GA carbonyl-C2-H, UU imino-4-carbonyl 2 carbonyl-C5-H, AC amino(A) N3(C)-carbonyl, GC imino amino-carbonyl, Gpsi imino-2-carbonyl amino-2-carbonyl, and GU imino amino-2-carbonyl base pairs.

By “histone deacetylase” or “HDAC” as used herein is meant, any histone deacetylate protein, peptide, or polypeptide having HDAC activity (e.g., HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7) such as encoded by HDAC or Sirtuin Genbank Accession Nos. shown in Table I and in U.S. Ser. No. 10/923,536 and U.S. Ser. No. 10/923,536, both incorporated by reference herein. The term HDAC also refers to nucleic acid sequences encoding any HDAC protein, peptide, or polypeptide having HDAC activity. The term “HDAC” is also meant to include other HDAC encoding sequence, such as other histone deacetylase isoforms, mutant HDAC genes, splice variants of HDAC genes, HDAC gene polymorphisms, and non-coding or regulatory HDAC polynucleotide sequences.

By “target” as used herein is meant, any target protein, peptide, or polypeptide, such as encoded by Genbank Accession Nos. shown in U.S. Ser. No. 10/923,536 and U.S. Ser. No. 10/923,536, both incorporated by reference herein. The term “target” also refers to nucleic acid sequences or target polynucleotide sequence encoding any target protein, peptide, or polypeptide, such as proteins, peptides, or polypeptides encoded by sequences having Genbank Accession Nos. shown in U.S. Ser. No. 10/923,536 and U.S. Ser. No. 10/923,536. The term “target” is also meant to include other sequences, such as differing isoforms, mutant target genes, splice variants of target polynucleotides, target polymorphisms, and non-coding or regulatory polynucleotide sequences.

By “homologous sequence” is meant, a nucleotide sequence that is shared by one or more polynucleotide sequences, such as genes, gene transcripts and/or non-coding polynucleotides. For example, a homologous sequence can be a nucleotide sequence that is shared by two or more genes encoding related but different proteins, such as different members of a gene family, different protein epitopes, different protein isoforms or completely divergent genes, such as a cytokine and its corresponding receptors. A homologous sequence can be a nucleotide sequence that is shared by two or more non-coding polynucleotides, such as noncoding DNA or RNA, regulatory sequences, introns, and sites of transcriptional control or regulation. Homologous sequences can also include conserved sequence regions shared by more than one polynucleotide sequence. Homology does not need to be perfect homology (e.g., 100%), as partially homologous sequences are also contemplated by the instant invention (e.g., 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80% etc.).

By “conserved sequence region” is meant, a nucleotide sequence of one or more regions in a polynucleotide does not vary significantly between generations or from one biological system, subject, or organism to another biological system, subject, or organism. The polynucleotide can include both coding and non-coding DNA and RNA.

By “sense region” is meant a nucleotide sequence of a siNA molecule having complementarity to an antisense region of the siNA molecule. In addition, the sense region of a siNA molecule can comprise a nucleic acid sequence having homology with a target nucleic acid sequence.

By “antisense region” is meant a nucleotide sequence of a siNA molecule having complementarity to a target nucleic acid sequence. In addition, the antisense region of a siNA molecule can optionally comprise a nucleic acid sequence having complementarity to a sense region of the siNA molecule.

By “target nucleic acid” or “target polynucleotide” is meant any nucleic acid sequence whose expression or activity is to be modulated (e.g., HDAC). The target nucleic acid can be DNA or RNA. In one embodiment, a target nucleic acid of the invention is target HDAC RNA or HDAC DNA.

By “complementarity” is meant that a nucleic acid can form hydrogen bond(s) with another nucleic acid sequence by either traditional Watson-Crick or other non-traditional types as described herein. In one embodiment, a double stranded nucleic acid molecule of the invention, such as an siNA molecule, wherein each strand is between 15 and 30 nucleotides in length, comprises between about 10% and about 100% (e.g., about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) complementarity between the two strands of the double stranded nucleic acid molecule. In another embodiment, a double stranded nucleic acid molecule of the invention, such as an siNA molecule, where one strand is the sense strand and the other stand is the antisense strand, wherein each strand is between 15 and 30 nucleotides in length, comprises between at least about 10% and about 100% (e.g., at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) complementarity between the nucleotide sequence in the antisense strand of the double stranded nucleic acid molecule and the nucleotide sequence of its corresponding target nucleic acid molecule, such as a target RNA or target mRNA or viral RNA. In one embodiment, a double stranded nucleic acid molecule of the invention, such as an siNA molecule, where one strand comprises nucleotide sequence that is referred to as the sense region and the other strand comprises a nucleotide sequence that is referred to as the antisense region, wherein each strand is between 15 and 30 nucleotides in length, comprises between about 10% and about 100% (e.g., about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) complementarity between the sense region and the antisense region of the double stranded nucleic acid molecule. In reference to the nucleic molecules of the present invention, the binding free energy for a nucleic acid molecule with its complementary sequence is sufficient to allow the relevant function of the nucleic acid to proceed, e.g., RNAi activity. Determination of binding free energies for nucleic acid molecules is well known in the art (see, e.g., Turner et al., 1987, CSH Symp. Quant. Biol. LII pp. 123-133; Frier et al., 1986, Proc. Nat. Acad. Sci. USA 83:9373-9377; Turner et al., 1987, J. Am. Chem. Soc. 109:3783-3785). A percent complementarity indicates the percentage of contiguous residues in a nucleic acid molecule that can form hydrogen bonds (e.g., Watson-Crick base pairing) with a second nucleic acid sequence (e.g., 5, 6, 7, 8, 9, or 10 nucleotides out of a total of 10 nucleotides in the first oligonucleotide being based paired to a second nucleic acid sequence having 10 nucleotides represents 50%, 60%, 70%, 80%, 90%, and 100% complementary respectively). In one embodiment, a siNA molecule of the invention has perfect complementarity between the sense strand or sense region and the antisense strand or antisense region of the siNA molecule. In one embodiment, a siNA molecule of the invention is perfectly complementary to a corresponding target nucleic acid molecule. “Perfectly complementary” means that all the contiguous residues of a nucleic acid sequence will hydrogen bond with the same number of contiguous residues in a second nucleic acid sequence. In one embodiment, a siNA molecule of the invention comprises about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more) nucleotides that are complementary to one or more target nucleic acid molecules or a portion thereof. In one embodiment, a siNA molecule of the invention has partial complementarity (i.e., less than 100% complementarity) between the sense strand or sense region and the antisense strand or antisense region of the siNA molecule or between the antisense strand or antisense region of the siNA molecule and a corresponding target nucleic acid molecule. For example, partial complementarity can include various mismatches or non-based paired nucleotides (e.g., 1, 2, 3, 4, 5 or more mismatches or non-based paired nucleotides) within the siNA structure which can result in bulges, loops, or overhangs that result between the between the sense strand or sense region and the antisense strand or antisense region of the siNA molecule or between the antisense strand or antisense region of the siNA molecule and a corresponding target nucleic acid molecule.

In one embodiment, a double stranded nucleic acid molecule of the invention, such as siNA molecule, has perfect complementarity between the sense strand or sense region and the antisense strand or antisense region of the nucleic acid molecule. In one embodiment, double stranded nucleic acid molecule of the invention, such as siNA molecule, is perfectly complementary to a corresponding target nucleic acid molecule.

In one embodiment, double stranded nucleic acid molecule of the invention, such as siNA molecule, has partial complementarity (i.e., less than 100% complementarity) between the sense strand or sense region and the antisense strand or antisense region of the double stranded nucleic acid molecule or between the antisense strand or antisense region of the nucleic acid molecule and a corresponding target nucleic acid molecule. For example, partial complementarity can include various mismatches or non-base paired nucleotides (e.g., 1, 2, 3, 4, 5 or more mismatches or non-based paired nucleotides, such as nucleotide bulges) within the double stranded nucleic acid molecule, structure which can result in bulges, loops, or overhangs that result between the sense strand or sense region and the antisense strand or antisense region of the double stranded nucleic acid molecule or between the antisense strand or antisense region of the double stranded nucleic acid molecule and a corresponding target nucleic acid molecule.

In one embodiment, a double stranded nucleic acid molecule of the invention is a microRNA (miRNA). By “mircoRNA” or “miRNA” is meant, a small double stranded RNA that regulates the expression of target messenger RNAs either by mRNA cleavage, translational repression/inhibition or heterochromatic silencing (see for example Ambros, 2004, Nature, 431, 350-355; Bartel, 2004, Cell, 116, 281-297; Cullen, 2004, Virus Research., 102, 3-9; He et al., 2004, Nat. Rev. Genet., 5, 522-531; and Ying et al., 2004, Gene, 342, 25-28). In one embodiment, the microRNA of the invention, has partial complementarity (i.e., less than 100% complementarity) between the sense strand or sense region and the antisense strand or antisense region of the miRNA molecule or between the antisense strand or antisense region of the miRNA and a corresponding target nucleic acid molecule. For example, partial complementarity can include various mismatches or non-base paired nucleotides (e.g., 1, 2, 3, 4, 5 or more mismatches or non-based paired nucleotides, such as nucleotide bulges) within the double stranded nucleic acid molecule, structure which can result in bulges, loops, or overhangs that result between the sense strand or sense region and the antisense strand or antisense region of the miRNA or between the antisense strand or antisense region of the miRNA and a corresponding target nucleic acid molecule.

In one embodiment, siNA molecules of the invention that down regulate or reduce target gene expression are used for preventing or treating diseases, disorders, conditions, or traits in a subject or organism as described herein or otherwise known in the art.

By “proliferative disease” or “cancer” as used herein is meant, any disease, condition, trait, genotype or phenotype characterized by unregulated cell growth or replication as is known in the art; including leukemias, for example, acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphocytic leukemia (ALL), and chronic lymphocytic leukemia, AIDS related cancers such as Kaposi's sarcoma; breast cancers; bone cancers such as Osteosarcoma, Chondrosarcomas, Ewing's sarcoma, Fibrosarcomas, Giant cell tumors, Adamantinomas, and Chordomas; Brain cancers such as Meningiomas, Glioblastomas, Lower-Grade Astrocytomas, Oligodendrocytomas, Pituitary Tumors, Schwannomas, and Metastatic brain cancers; cancers of the head and neck including various lymphomas such as mantle cell lymphoma, non-Hodgkins lymphoma, adenoma, squamous cell carcinoma, laryngeal carcinoma, gallbladder and bile duct cancers, cancers of the retina such as retinoblastoma, cancers of the esophagus, gastric cancers, multiple myeloma, ovarian cancer, uterine cancer, thyroid cancer, testicular cancer, endometrial cancer, melanoma, colorectal cancer, lung cancer, bladder cancer, prostate cancer, lung cancer (including non-small cell lung carcinoma), pancreatic cancer, sarcomas, Wilms' tumor, cervical cancer, head and neck cancer, skin cancers, nasopharyngeal carcinoma, liposarcoma, epithelial carcinoma, renal cell carcinoma, gallbladder adeno carcinoma, parotid adenocarcinoma, endometrial sarcoma, multidrug resistant cancers; and proliferative diseases and conditions, such as neovascularization associated with tumor angiogenesis, and ocular diseases such as macular degeneration (e.g., wet/dry AMD), corneal neovascularization, diabetic retinopathy, neovascular glaucoma, myopic degeneration and other proliferative diseases and conditions such as restenosis and polycystic kidney disease, and any other cancer or proliferative disease, condition, trait, genotype or phenotype that can respond to the modulation of disease related gene expression in a cell or tissue, alone or in combination with other therapies.

By “ocular disease” as used herein is meant, any disease, condition, trait, genotype or phenotype of the eye and related structures as is known in the art, such as Cystoid Macular Edema, Asteroid Hyalosis, Pathological Myopia and Posterior Staphyloma, Toxocariasis (Ocular Larva Migrans), Retinal Vein Occlusion, Posterior Vitreous Detachment, Tractional Retinal Tears, Epiretinal Membrane, Diabetic Retinopathy, Lattice Degeneration, Retinal Vein Occlusion, Retinal Artery Occlusion, Macular Degeneration (e.g., age related macular degeneration such as wet AMD or dry AMD), Toxoplasmosis, Choroidal Melanoma, Acquired Retinoschisis, Hollenhorst Plaque, Idiopathic Central Serous Chorioretinopathy, Macular Hole, Presumed Ocular Histoplasmosis Syndrome, Retinal Macroaneursym, Retinitis Pigmentosa, Retinal Detachment, Hypertensive Retinopathy, Retinal Pigment Epithelium (RPE) Detachment, Papillophlebitis, Ocular Ischemic Syndrome, Coats' Disease, Leber's Miliary Aneurysm, Conjunctival Neoplasms, Allergic Conjunctivitis, Vernal Conjunctivitis, Acute Bacterial Conjunctivitis, Allergic Conjunctivitis & Vernal Keratoconjunctivitis, Viral Conjunctivitis, Bacterial Conjunctivitis, Chlamydial & Gonococcal Conjunctivitis, Conjunctival Laceration, Episcleritis, Scleritis, Pingueculitis, Pterygium, Superior Limbic Keratoconjunctivitis (SLK of Theodore), Toxic Conjunctivitis, Conjunctivitis with Pseudomembrane, Giant Papillary Conjunctivitis, Terrien's Marginal Degeneration, Acanthamoeba Keratitis, Fungal Keratitis, Filamentary Keratitis, Bacterial Keratitis, Keratitis Sicca/Dry Eye Syndrome, Bacterial Keratitis, Herpes Simplex Keratitis, Sterile Corneal Infiltrates, Phlyctenulosis, Corneal Abrasion & Recurrent Corneal Erosion, Corneal Foreign Body, Chemical Burs, Epithelial Basement Membrane Dystrophy (EBMD), Thygeson's Superficial Punctate Keratopathy, Corneal Laceration, Salzmann's Nodular Degeneration, Fuchs' Endothelial Dystrophy, Crystalline Lens Subluxation, Ciliary-Block Glaucoma, Primary Open-Angle Glaucoma, Pigment Dispersion Syndrome and Pigmentary Glaucoma, Pseudoexfoliation Syndrom and Pseudoexfoliative Glaucoma, Anterior Uveitis, Primary Open Angle Glaucoma, Uveitic Glaucoma & Glaucomatocyclitic Crisis, Pigment Dispersion Syndrome & Pigmentary Glaucoma, Acute Angle Closure Glaucoma, Anterior Uveitis, Hyphema, Angle Recession Glaucoma, Lens Induced Glaucoma, Pseudoexfoliation Syndrome and Pseudoexfoliative Glaucoma, Axenfeld-Rieger Syndrome, Neovascular Glaucoma, Pars Planitis, Choroidal Rupture, Duane's Retraction Syndrome, Toxic/Nutritional Optic Neuropathy, Aberrant Regeneration of Cranial Nerve III, Intracranial Mass Lesions, Carotid-Cavernous Sinus Fistula, Anterior Ischemic Optic Neuropathy, Optic Disc Edema & Papilledema, Cranial Nerve III Palsy, Cranial Nerve IV Palsy, Cranial Nerve VI Palsy, Cranial Nerve VII (Facial Nerve) Palsy, Horner's Syndrome, Internuclear Ophthalmoplegia, Optic Nerve Head Hypoplasia, Optic Pit, Tonic Pupil, Optic Nerve Head Drusen, Demyelinating Optic Neuropathy (Optic Neuritis, Retrobulbar Optic Neuritis), Amaurosis Fugax and Transient Ischemic Attack, Pseudotumor Cerebri, Pituitary Adenoma, Molluscum Contagiosum, Canaliculitis, Verruca and Papilloma, Pediculosis and Pthiriasis, Blepharitis, Hordeolum, Preseptal Cellulitis, Chalazion, Basal Cell Carcinoma, Herpes Zoster Ophthalmicus, Pediculosis & Phthiriasis, Blow-out Fracture, Chronic Epiphora, Dacryocystitis, Herpes Simplex Blepharitis, Orbital Cellulitis, Senile Entropion, and Squamous Cell Carcinoma.

In one embodiment of the present invention, each sequence of a siNA molecule of the invention is independently about 15 to about 30 nucleotides in length, in specific embodiments about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. In another embodiment, the siNA duplexes of the invention independently comprise about 15 to about 30 base pairs (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30). In another embodiment, one or more strands of the siNA molecule of the invention independently comprises about 15 to about 30 nucleotides (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) that are complementary to a target nucleic acid molecule. In yet another embodiment, siNA molecules of the invention comprising hairpin or circular structures are about 35 to about 55 (e.g., about 35, 40, 45, 50 or 55) nucleotides in length, or about 38 to about 44 (e.g., about 38, 39, 40, 41, 42, 43, or 44) nucleotides in length and comprising about 15 to about 25 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) base pairs. Exemplary siNA molecules of the invention are shown in Table II and/or FIGS. 4-5.

As used herein “cell” is used in its usual biological sense, and does not refer to an entire multicellular organism, e.g., specifically does not refer to a human. The cell can be present in an organism, e.g., birds, plants and mammals such as humans, cows, sheep, apes, monkeys, swine, dogs, and cats. The cell can be prokaryotic (e.g., bacterial cell) or eukaryotic (e.g., mammalian or plant cell). The cell can be of somatic or germ line origin, totipotent or pluripotent, dividing or non-dividing. The cell can also be derived from or can comprise a gamete or embryo, a stem cell, or a fully differentiated cell.

The siNA molecules of the invention are added directly, or can be complexed with cationic lipids, packaged within liposomes, or otherwise delivered to target cells or tissues. The nucleic acid or nucleic acid complexes can be locally administered to relevant tissues ex vivo, or in vivo through local delivery to the lung, with or without their incorporation in biopolymers. In particular embodiments, the nucleic acid molecules of the invention comprise sequences shown in Tables II-III and/or FIGS. 4-5. Examples of such nucleic acid molecules consist essentially of sequences defined in these tables and figures. Furthermore, the chemically modified constructs described in Table IV can be applied to any siNA sequence of the invention.

In another aspect, the invention provides mammalian cells containing one or more siNA molecules of this invention. The one or more siNA molecules can independently be targeted to the same or different sites.

By “RNA” is meant a molecule comprising at least one ribonucleotide residue. By “ribonucleotide” is meant a nucleotide with a hydroxyl group at the 2′ position of a β-D-ribofuranose moiety. The terms include double-stranded RNA, single-stranded RNA, isolated RNA such as partially purified RNA, essentially pure RNA, synthetic RNA, recombinantly produced RNA, as well as altered RNA that differs from naturally occurring RNA by the addition, deletion, substitution and/or alteration of one or more nucleotides. Such alterations can include addition of non-nucleotide material, such as to the end(s) of the siNA or internally, for example at one or more nucleotides of the RNA. Nucleotides in the RNA molecules of the instant invention can also comprise non-standard nucleotides, such as non-naturally occurring nucleotides or chemically synthesized nucleotides or deoxynucleotides. These altered RNAs can be referred to as analogs or analogs of naturally-occurring RNA.

By “subject” is meant an organism, which is a donor or recipient of explanted cells or the cells themselves. “Subject” also refers to an organism to which the nucleic acid molecules of the invention can be administered. A subject can be a mammal or mammalian cells, including a human or human cells.

By “chemical modification” as used herein is meant any modification of chemical structure of the nucleotides that differs from nucleotides of native siRNA or RNA. The term “chemical modification” encompasses the addition, substitution, or modification of native siRNA or RNA nucleosides and nucleotides with modified nucleosides and modified nucleotides as described herein or as is otherwise known in the art. Non-limiting examples of such chemical modifications include without limitation phosphorothioate internucleotide linkages, 2′-deoxyribonucleotides, 2′-O-methyl ribonucleotides, 2′-deoxy-2′-fluoro ribonucleotides, 4′-thio ribonucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides (see for example U.S. Ser. No. 10/981,966 filed Nov. 5, 2004, incorporated by reference herein), “universal base” nucleotides, “acyclic” nucleotides, 5-C-methyl nucleotides, terminal glyceryl and/or inverted deoxy abasic residue incorporation, or a modification having any of Formulae I-VII herein.

The term “phosphorothioate” as used herein refers to an internucleotide linkage having Formula I, wherein Z and/or W comprise a sulfur atom. Hence, the term phosphorothioate refers to both phosphorothioate and phosphorodithioate internucleotide linkages.

The term “phosphonoacetate” as used herein refers to an internucleotide linkage having Formula I, wherein Z and/or W comprise an acetyl or protected acetyl group.

The term “thiophosphonoacetate” as used herein refers to an internucleotide linkage having Formula I, wherein Z comprises an acetyl or protected acetyl group and W comprises a sulfur atom or alternately W comprises an acetyl or protected acetyl group and Z comprises a sulfur atom.

The term “universal base” as used herein refers to nucleotide base analogs that form base pairs with each of the natural DNA/RNA bases with little discrimination between them. Non-limiting examples of universal bases include C-phenyl, C-naphthyl and other aromatic derivatives, inosine, azole carboxamides, and nitroazole derivatives such as 3-nitropyrrole, 4-nitroindole, 5-nitroindole, and 6-nitroindole as known in the art (see for example Loakes, 2001, Nucleic Acids Research, 29, 2437-2447).

The term “acyclic nucleotide” as used herein refers to any nucleotide having an acyclic ribose sugar, for example where any of the ribose carbons (C1, C2, C3, C4, or C5), are independently or in combination absent from the nucleotide.

The nucleic acid molecules of the instant invention, individually, or in combination or in conjunction with other drugs, can be used to for preventing or treating diseases, disorders, conditions, and traits described herein or otherwise known in the art, in a subject or organism.

In one embodiment, the siNA molecules of the invention can be administered to a subject or can be administered to other appropriate cells evident to those skilled in the art, individually or in combination with one or more drugs under conditions suitable for the treatment.

In a further embodiment, the siNA molecules can be used in combination with other known treatments to prevent or treat in a subject or organism. For example, the described molecules could be used in combination with one or more known compounds, treatments, or procedures to prevent or treat diseases, disorders, conditions, and traits described herein in a subject or organism as are known in the art.

In one embodiment, the invention features an expression vector comprising a nucleic acid sequence encoding at least one siNA molecule of the invention, in a manner which allows expression of the siNA molecule. For example, the vector can contain sequence(s) encoding both strands of a siNA molecule comprising a duplex. The vector can also contain sequence(s) encoding a single nucleic acid molecule that is self-complementary and thus forms a siNA molecule. Non-limiting examples of such expression vectors are described in Paul et al., 2002, Nature Biotechnology, 19, 505; Miyagishi and Taira, 2002, Nature Biotechnology, 19, 497; Lee et al., 2002, Nature Biotechnology, 19, 500; and Novina et al., 2002, Nature Medicine, advance online publication doi:10.1038/nm725.

In another embodiment, the invention features a mammalian cell, for example, a human cell, including an expression vector of the invention.

In yet another embodiment, the expression vector of the invention comprises a sequence for a siNA molecule having complementarity to a RNA molecule referred to by a Genbank Accession numbers, for example Genbank Accession Nos. shown in Table I herein or in U.S. Ser. No. 10/923,536 and U.S. Ser. No. 10/923,536, both incorporated by reference herein.

In one embodiment, an expression vector of the invention comprises a nucleic acid sequence encoding two or more siNA molecules, which can be the same or different.

In another aspect of the invention, siNA molecules that interact with target RNA molecules and down-regulate gene encoding target RNA molecules (for example target RNA molecules referred to by Genbank Accession numbers herein) are expressed from transcription units inserted into DNA or RNA vectors. The recombinant vectors can be DNA plasmids or viral vectors. siNA expressing viral vectors can be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. The recombinant vectors capable of expressing the siNA molecules can be delivered as described herein, and persist in target cells. Alternatively, viral vectors can be used that provide for transient expression of siNA molecules. Such vectors can be repeatedly administered as necessary. Once expressed, the siNA molecules bind and down-regulate gene function or expression via RNA interference (RNAi). Delivery of siNA expressing vectors can be systemic, such as by intravenous or intramuscular administration, by administration to target cells ex-planted from a subject followed by reintroduction into the subject, or by any other means that would allow for introduction into the desired target cell.

By “vectors” is meant any nucleic acid- and/or viral-based technique used to deliver a desired nucleic acid.

Other features and advantages of the invention will be apparent from the following description of the preferred embodiments thereof, and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a non-limiting example of a scheme for the synthesis of siNA molecules. The complementary siNA sequence strands, strand 1 and strand 2, are synthesized in tandem and are connected by a cleavable linkage, such as a nucleotide succinate or abasic succinate, which can be the same or different from the cleavable linker used for solid phase synthesis on a solid support. The synthesis can be either solid phase or solution phase, in the example shown, the synthesis is a solid phase synthesis. The synthesis is performed such that a protecting group, such as a dimethoxytrityl group, remains intact on the terminal nucleotide of the tandem oligonucleotide. Upon cleavage and deprotection of the oligonucleotide, the two siNA strands spontaneously hybridize to form a siNA duplex, which allows the purification of the duplex by utilizing the properties of the terminal protecting group, for example by applying a trityl on purification method wherein only duplexes/oligonucleotides with the terminal protecting group are isolated.

FIG. 2 shows a MALDI-TOF mass spectrum of a purified siNA duplex synthesized by a method of the invention. The two peaks shown correspond to the predicted mass of the separate siNA sequence strands. This result demonstrates that the siNA duplex generated from tandem synthesis can be purified as a single entity using a simple trityl-on purification methodology.

FIG. 3 shows a non-limiting proposed mechanistic representation of target RNA degradation involved in RNAi. Double-stranded RNA (dsRNA), which is generated by RNA-dependent RNA polymerase (RdRP) from foreign single-stranded RNA, for example viral, transposon, or other exogenous RNA, activates the DICER enzyme that in turn generates siNA duplexes. Alternately, synthetic or expressed siNA can be introduced directly into a cell by appropriate means. An active siNA complex forms which recognizes a target RNA, resulting in degradation of the target RNA by the RISC endonuclease complex or in the synthesis of additional RNA by RNA-dependent RNA polymerase (RdRP), which can activate DICER and result in additional siNA molecules, thereby amplifying the RNAi response.

FIG. 4A-F shows non-limiting examples of chemically-modified siNA constructs of the present invention. In the figure, N stands for any nucleotide (adenosine, guanosine, cytosine, uridine, or optionally thymidine, for example thymidine can be substituted in the overhanging regions designated by parenthesis (N N). Various modifications are shown for the sense and antisense strands of the siNA constructs.

FIG. 4A: The sense strand comprises 21 nucleotides wherein the two terminal 3′-nucleotides are optionally base paired and wherein all nucleotides present are ribonucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, and wherein all nucleotides present are ribonucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the antisense strand.

FIG. 4B: The sense strand comprises 21 nucleotides wherein the two terminal 3′-nucleotides are optionally base paired and wherein all pyrimidine nucleotides that may be present are 2′deoxy-2′-fluoro modified nucleotides and all purine nucleotides that may be present are 2′-O-methyl modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety and wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides and all purine nucleotides that may be present are 2′-O-methyl modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the sense and antisense strand.

FIG. 4C: The sense strand comprises 21 nucleotides having 5′- and 3′-terminal cap moieties wherein the two terminal 3′-nucleotides are optionally base paired and wherein all pyrimidine nucleotides that may be present are 2′-O-methyl or 2′-deoxy-2′-fluoro modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety and wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the antisense strand.

FIG. 4D: The sense strand comprises 21 nucleotides having 5′- and 3′-terminal cap moieties wherein the two terminal 3′-nucleotides are optionally base paired and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein and wherein and all purine nucleotides that may be present are 2′-deoxy nucleotides. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety and wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides and all purine nucleotides that may be present are 2′-O-methyl modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the antisense strand.

FIG. 4E: The sense strand comprises 21 nucleotides having 5′- and 3′-terminal cap moieties wherein the two terminal 3′-nucleotides are optionally base paired and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety and wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides and all purine nucleotides that may be present are 2′-O-methyl modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the antisense strand.

FIG. 4F: The sense strand comprises 21 nucleotides having 5′- and 3′-terminal cap moieties wherein the two terminal 3′-nucleotides are optionally base paired and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein and wherein and all purine nucleotides that may be present are 2′-deoxy nucleotides. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety and wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, and having one 3′-terminal phosphorothioate internucleotide linkage and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides and all purine nucleotides that may be present are 2′-deoxy nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the antisense strand. The antisense strand of constructs A-F comprise sequence complementary to any target nucleic acid sequence of the invention. Furthermore, when a glyceryl moiety (L) is present at the 3′-end of the antisense strand for any construct shown in FIG. 4 A-F, the modified internucleotide linkage is optional.

FIG. 5A-F shows non-limiting examples of specific chemically-modified siNA sequences of the invention. A-F applies the chemical modifications described in FIG. 4A-F to an exemplary HDAC 11 siNA sequence. Such chemical modifications can be applied to any target polynucleotide sequence.

FIG. 6A-B shows non-limiting examples of different siNA constructs of the invention.

The examples shown in FIG. 6A (constructs 1, 2, and 3) have 19 representative base pairs; however, different embodiments of the invention include any number of base pairs described herein. Bracketed regions represent nucleotide overhangs, for example, comprising about 1, 2, 3, or 4 nucleotides in length, preferably about 2 nucleotides. Constructs 1 and 2 can be used independently for RNAi activity. Construct 2 can comprise a polynucleotide or non-nucleotide linker, which can optionally be designed as a biodegradable linker. In one embodiment, the loop structure shown in construct 2 can comprise a biodegradable linker that results in the formation of construct 1 in vivo and/or in vitro. In another example, construct 3 can be used to generate construct 2 under the same principle wherein a linker is used to generate the active siNA construct 2 in vivo and/or in vitro, which can optionally utilize another biodegradable linker to generate the active siNA construct 1 in vivo and/or in vitro. As such, the stability and/or activity of the siNA constructs can be modulated based on the design of the siNA construct for use in vivo or in vitro and/or in vitro.

The examples shown in FIG. 6B represent different variations of double stranded nucleic acid molecule of the invention, such as microRNA, that can include overhangs, bulges, loops, and stem-loops resulting from partial complementarity. Such motifs having bulges, loops, and stem-loops are generally characteristics of miRNA. The bulges, loops, and stem-loops can result from any degree of partial complementarity, such as mismatches or bulges of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more nucleotides in one or both strands of the double stranded nucleic acid molecule of the invention.

FIG. 7A-C is a diagrammatic representation of a scheme utilized in generating an expression cassette to generate siNA hairpin constructs.

FIG. 7A: A DNA oligomer is synthesized with a 5′-restriction site (R1) sequence followed by a region having sequence identical (sense region of siNA) to a predetermined target sequence, wherein the sense region comprises, for example, about 19, 20, 21, or 22 nucleotides (N) in length, which is followed by a loop sequence of defined sequence (X), comprising, for example, about 3 to about 10 nucleotides.

FIG. 7B: The synthetic construct is then extended by DNA polymerase to generate a hairpin structure having self-complementary sequence that will result in a siNA transcript having specificity for a target sequence and having self-complementary sense and antisense regions.

FIG. 7C: The construct is heated (for example to about 95° C.) to linearize the sequence, thus allowing extension of a complementary second DNA strand using a primer to the 3′-restriction sequence of the first strand. The double-stranded DNA is then inserted into an appropriate vector for expression in cells. The construct can be designed such that a 3′-terminal nucleotide overhang results from the transcription, for example, by engineering restriction sites and/or utilizing a poly-U termination region as described in Paul et al., 2002, Nature Biotechnology, 29, 505-508.

FIG. 8A-C is a diagrammatic representation of a scheme utilized in generating an expression cassette to generate double-stranded siNA constructs.

FIG. 8A: A DNA oligomer is synthesized with a 5′-restriction (R1) site sequence followed by a region having sequence identical (sense region of siNA) to a predetermined target sequence, wherein the sense region comprises, for example, about 19, 20, 21, or 22 nucleotides (N) in length, and which is followed by a 3′-restriction site (R2) which is adjacent to a loop sequence of defined sequence (X).

FIG. 8B: The synthetic construct is then extended by DNA polymerase to generate a hairpin structure having self-complementary sequence.

FIG. 8C: The construct is processed by restriction enzymes specific to R1 and R2 to generate a double-stranded DNA which is then inserted into an appropriate vector for expression in cells. The transcription cassette is designed such that a U6 promoter region flanks each side of the dsDNA which generates the separate sense and antisense strands of the siNA. Poly T termination sequences can be added to the constructs to generate U overhangs in the resulting transcript.

FIG. 9A-E is a diagrammatic representation of a method used to determine target sites for siNA mediated RNAi within a particular target nucleic acid sequence, such as messenger RNA.

FIG. 9A: A pool of siNA oligonucleotides are synthesized wherein the antisense region of the siNA constructs has complementarity to target sites across the target nucleic acid sequence, and wherein the sense region comprises sequence complementary to the antisense region of the siNA.

FIGS. 9B&C: (FIG. 9B) The sequences are pooled and are inserted into vectors such that (FIG. 9C) transfection of a vector into cells results in the expression of the siNA.

FIG. 9D: Cells are sorted based on phenotypic change that is associated with modulation of the target nucleic acid sequence.

FIG. 9E: The siNA is isolated from the sorted cells and is sequenced to identify efficacious target sites within the target nucleic acid sequence.

FIG. 10 shows non-limiting examples of different stabilization chemistries (1-10) that can be used, for example, to stabilize the 3′-end of siNA sequences of the invention, including (1) [3-3′]-inverted deoxyribose; (2) deoxyribonucleotide; (3) [5′-3′]-3′ deoxyribonucleotide; (4) [5′-3′]-ribonucleotide; (5) [5′-3′]-3′-O-methyl ribonucleotide; (6) 3′-glyceryl; (7) [3′-5′]-3′-deoxyribonucleotide; (8) [3′-3′]-deoxyribonucleotide; (9) [5′-2′]-deoxyribonucleotide; and (10) [5-3′]-dideoxyribonucleotide. In addition to modified and unmodified backbone chemistries indicated in the figure, these chemistries can be combined with different backbone modifications as described herein, for example, backbone modifications having Formula I. In addition, the 2′-deoxy nucleotide shown 5′ to the terminal modifications shown can be another modified or unmodified nucleotide or non-nucleotide described herein, for example modifications having any of Formulae I-VII or any combination thereof.

FIG. 11 shows a non-limiting example of a strategy used to identify chemically modified siNA constructs of the invention that are nuclease resistance while preserving the ability to mediate RNAi activity. Chemical modifications are introduced into the siNA construct based on educated design parameters (e.g. introducing 2′-mofications, base modifications, backbone modifications, terminal cap modifications etc). The modified construct in tested in an appropriate system (e.g. human serum for nuclease resistance, shown, or an animal model for PK/delivery parameters). In parallel, the siNA construct is tested for RNAi activity, for example in a cell culture system such as a luciferase reporter assay). Lead siNA constructs are then identified which possess a particular characteristic while maintaining RNAi activity, and can be further modified and assayed once again. This same approach can be used to identify siNA-conjugate molecules with improved pharmacokinetic profiles, delivery, and RNAi activity.

FIG. 12 shows non-limiting examples of phosphorylated siNA molecules of the invention, including linear and duplex constructs and asymmetric derivatives thereof.

FIG. 13 shows non-limiting examples of chemically modified terminal phosphate groups of the invention.

FIG. 14A shows a non-limiting example of methodology used to design self complementary DFO constructs utilizing palindrome and/or repeat nucleic acid sequences that are identified in a target nucleic acid sequence. (i) A palindrome or repeat sequence is identified in a nucleic acid target sequence. (ii) A sequence is designed that is complementary to the target nucleic acid sequence and the palindrome sequence. (iii) An inverse repeat sequence of the non-palindrome/repeat portion of the complementary sequence is appended to the 3′-end of the complementary sequence to generate a self complementary DFO molecule comprising sequence complementary to the nucleic acid target. (iv) The DFO molecule can self-assemble to form a double stranded oligonucleotide. FIG. 14B shows a non-limiting representative example of a duplex forming oligonucleotide sequence. FIG. 14C shows a non-limiting example of the self assembly schematic of a representative duplex forming oligonucleotide sequence. FIG. 14D shows a non-limiting example of the self assembly schematic of a representative duplex forming oligonucleotide sequence followed by interaction with a target nucleic acid sequence resulting in modulation of gene expression.

FIG. 15 shows a non-limiting example of the design of self complementary DFO constructs utilizing palindrome and/or repeat nucleic acid sequences that are incorporated into the DFO constructs that have sequence complementary to any target nucleic acid sequence of interest. Incorporation of these palindrome/repeat sequences allow the design of DFO constructs that form duplexes in which each strand is capable of mediating modulation of target gene expression, for example by RNAi. First, the target sequence is identified. A complementary sequence is then generated in which nucleotide or non-nucleotide modifications (shown as X or Y) are introduced into the complementary sequence that generate an artificial palindrome (shown as XYXYXY in the Figure). An inverse repeat of the non-palindrome/repeat complementary sequence is appended to the 3′-end of the complementary sequence to generate a self complementary DFO comprising sequence complementary to the nucleic acid target. The DFO can self-assemble to form a double stranded oligonucleotide.

FIG. 16 shows non-limiting examples of multifunctional siNA molecules of the invention comprising two separate polynucleotide sequences that are each capable of mediating RNAi directed cleavage of differing target nucleic acid sequences. FIG. 16A shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first and second complementary regions are situated at the 3′-ends of each polynucleotide sequence in the multifunctional siNA. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences. FIG. 16B shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first and second complementary regions are situated at the 5′-ends of each polynucleotide sequence in the multifunctional siNA. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences.

FIG. 17 shows non-limiting examples of multifunctional siNA molecules of the invention comprising a single polynucleotide sequence comprising distinct regions that are each capable of mediating RNAi directed cleavage of differing target nucleic acid sequences. FIG. 17A shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the second complementary region is situated at the 3′-end of the polynucleotide sequence in the multifunctional siNA. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences. FIG. 17B shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first complementary region is situated at the 5′-end of the polynucleotide sequence in the multifunctional siNA. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences. In one embodiment, these multifunctional siNA constructs are processed in vivo or in vitro to generate multifunctional siNA constructs as shown in FIG. 16.

FIG. 18 shows non-limiting examples of multifunctional siNA molecules of the invention comprising two separate polynucleotide sequences that are each capable of mediating RNAi directed cleavage of differing target nucleic acid sequences and wherein the multifunctional siNA construct further comprises a self complementary, palindrome, or repeat region, thus enabling shorter bifuctional siNA constructs that can mediate RNA interference against differing target nucleic acid sequences. FIG. 18A shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first and second complementary regions are situated at the 3′-ends of each polynucleotide sequence in the multifunctional siNA, and wherein the first and second complementary regions further comprise a self complementary, palindrome, or repeat region. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences. FIG. 18B shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first and second complementary regions are situated at the 5′-ends of each polynucleotide sequence in the multifunctional siNA, and wherein the first and second complementary regions further comprise a self complementary, palindrome, or repeat region. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences.

FIG. 19 shows non-limiting examples of multifunctional siNA molecules of the invention comprising a single polynucleotide sequence comprising distinct regions that are each capable of mediating RNAi directed cleavage of differing target nucleic acid sequences and wherein the multifunctional siNA construct further comprises a self complementary, palindrome, or repeat region, thus enabling shorter bifuctional siNA constructs that can mediate RNA interference against differing target nucleic acid sequences. FIG. 19A shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the second complementary region is situated at the 3′-end of the polynucleotide sequence in the multifunctional siNA, and wherein the first and second complementary regions further comprise a self complementary, palindrome, or repeat region. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences. FIG. 19B shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first complementary region is situated at the 5′-end of the polynucleotide sequence in the multifunctional siNA, and wherein the first and second complementary regions further comprise a self complementary, palindrome, or repeat region. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences. In one embodiment, these multifunctional siNA constructs are processed in vivo or in vitro to generate multifunctional siNA constructs as shown in FIG. 18.

FIG. 20 shows a non-limiting example of how multifunctional siNA molecules of the invention can target two separate target nucleic acid molecules, such as separate RNA molecules encoding differing proteins, for example, a cytokine and its corresponding receptor, differing viral strains, a virus and a cellular protein involved in viral infection or replication, or differing proteins involved in a common or divergent biologic pathway that is implicated in the maintenance of progression of disease. Each strand of the multifunctional siNA construct comprises a region having complementarity to separate target nucleic acid molecules. The multifunctional siNA molecule is designed such that each strand of the siNA can be utilized by the RISC complex to initiate RNA interference mediated cleavage of its corresponding target. These design parameters can include destabilization of each end of the siNA construct (see for example Schwarz et al., 2003, Cell, 115, 199-208). Such destabilization can be accomplished for example by using guanosine-cytidine base pairs, alternate base pairs (e.g., wobbles), or destabilizing chemically modified nucleotides at terminal nucleotide positions as is known in the art.

FIG. 21 shows a non-limiting example of how multifunctional siNA molecules of the invention can target two separate target nucleic acid sequences within the same target nucleic acid molecule, such as alternate coding regions of a RNA, coding and non-coding regions of a RNA, or alternate splice variant regions of a RNA. Each strand of the multifunctional siNA construct comprises a region having complementarity to the separate regions of the target nucleic acid molecule. The multifunctional siNA molecule is designed such that each strand of the siNA can be utilized by the RISC complex to initiate RNA interference mediated cleavage of its corresponding target region. These design parameters can include destabilization of each end of the siNA construct (see for example Schwarz et al., 2003, Cell, 115, 199-208). Such destabilization can be accomplished for example by using guanosine-cytidine base pairs, alternate base pairs (e.g., wobbles), or destabilizing chemically modified nucleotides at terminal nucleotide positions as is known in the art.

FIG. 22(A-H) shows non-limiting examples of tethered multifunctional siNA constructs of the invention. In the examples shown, a linker (e.g., nucleotide or non-nucleotide linker) connects two siNA regions (e.g., two sense, two antisense, or alternately a sense and an antisense region together. Separate sense (or sense and antisense) sequences corresponding to a first target sequence and second target sequence are hybridized to their corresponding sense and/or antisense sequences in the multifunctional siNA. In addition, various conjugates, ligands, aptamers, polymers or reporter molecules can be attached to the linker region for selective or improved delivery and/or pharmacokinetic properties.

FIG. 23 shows a non-limiting example of various dendrimer based multifunctional siNA designs.

FIG. 24 shows a non-limiting example of various supramolecular multifunctional siNA designs.

FIG. 25 shows a non-limiting example of a dicer enabled multifunctional siNA design using a 30 nucleotide precursor siNA construct. A 30 base pair duplex is cleaved by Dicer into 22 and 8 base pair products from either end (8 b.p. fragments not shown). For ease of presentation the overhangs generated by dicer are not shown—but can be compensated for. Three targeting sequences are shown. The required sequence identity overlapped is indicated by grey boxes. The N's of the parent 30 b.p. siNA are suggested sites of 2′-OH positions to enable Dicer cleavage if this is tested in stabilized chemistries. Note that processing of a 30mer duplex by Dicer RNase III does not give a precise 22+8 cleavage, but rather produces a series of closely related products (with 22+8 being the primary site). Therefore, processing by Dicer will yield a series of active siNAs.

FIG. 26 shows a non-limiting example of a dicer enabled multifunctional siNA design using a 40 nucleotide precursor siNA construct. A 40 base pair duplex is cleaved by Dicer into 20 base pair products from either end. For ease of presentation the overhangs generated by dicer are not shown—but can be compensated for. Four targeting sequences are shown. The target sequences having homology are enclosed by boxes. This design format can be extended to larger RNAs. If chemically stabilized siNAs are bound by Dicer, then strategically located ribonucleotide linkages can enable designer cleavage products that permit our more extensive repertoire of multiifunctional designs. For example cleavage products not limited to the Dicer standard of approximately 22-nucleotides can allow multifunctional siNA constructs with a target sequence identity overlap ranging from, for example, about 3 to about 15 nucleotides.

FIG. 27 shows a non-limiting example of additional multifunctional siNA construct designs of the invention. In one example, a conjugate, ligand, aptamer, label, or other moiety is attached to a region of the multifunctional siNA to enable improved delivery or pharmacokinetic profiling.

FIG. 28 shows a non-limiting example of additional multifunctional siNA construct designs of the invention. In one example, a conjugate, ligand, aptamer, label, or other moiety is attached to a region of the multifunctional siNA to enable improved delivery or pharmacokinetic profiling.

FIG. 29 shows a non-limiting example of a cholesterol linked phosphoramidite that can be used to synthesize cholesterol conjugated siNA molecules of the invention. An example is shown with the cholesterol moiety linked to the 5′-end of the sense strand of a siNA molecule.

DETAILED DESCRIPTION OF THE INVENTION

Mechanism of Action of Nucleic Acid Molecules of the Invention

The discussion that follows discusses the proposed mechanism of RNA interference mediated by short interfering RNA as is presently known, and is not meant to be limiting and is not an admission of prior art. Applicant demonstrates herein that chemically-modified short interfering nucleic acids possess similar or improved capacity to mediate RNAi as do siRNA molecules and are expected to possess improved stability and activity in vivo; therefore, this discussion is not meant to be limiting only to siRNA and can be applied to siNA as a whole. By “improved capacity to mediate RNAi” or “improved RNAi activity” is meant to include RNAi activity measured in vitro and/or in vivo where the RNAi activity is a reflection of both the ability of the siNA to mediate RNAi and the stability of the siNAs of the invention. In this invention, the product of these activities can be increased in vitro and/or in vivo compared to an all RNA siRNA or a siNA containing a plurality of ribonucleotides. In some cases, the activity or stability of the siNA molecule can be decreased (i.e., less than ten-fold), but the overall activity of the siNA molecule is enhanced in vitro and/or in vivo.

RNA interference refers to the process of sequence specific post-transcriptional gene silencing in animals mediated by short interfering RNAs (siRNAs) (Fire et al., 1998, Nature, 391, 806). The corresponding process in plants is commonly referred to as post-transcriptional gene silencing or RNA silencing and is also referred to as quelling in fungi. The process of post-transcriptional gene silencing is thought to be an evolutionarily-conserved cellular defense mechanism used to prevent the expression of foreign genes which is commonly shared by diverse flora and phyla (Fire et al., 1999, Trends Genet., 15, 358). Such protection from foreign gene expression may have evolved in response to the production of double-stranded RNAs (dsRNAs) derived from viral infection or the random integration of transposon elements into a host genome via a cellular response that specifically destroys homologous single-stranded RNA or viral genomic RNA. The presence of dsRNA in cells triggers the RNAi response though a mechanism that has yet to be fully characterized. This mechanism appears to be different from the interferon response that results from dsRNA-mediated activation of protein kinase PKR and 2′,5′-oligoadenylate synthetase resulting in non-specific cleavage of mRNA by ribonuclease L.

The presence of long dsRNAs in cells stimulates the activity of a ribonuclease III enzyme referred to as Dicer. Dicer is involved in the processing of the dsRNA into short pieces of dsRNA known as short interfering RNAs (siRNAs) (Berstein et al., 2001, Nature, 409, 363). Short interfering RNAs derived from Dicer activity are typically about 21 to about 23 nucleotides in length and comprise about 19 base pair duplexes. Dicer has also been implicated in the excision of 21- and 22-nucleotide small temporal RNAs (stRNAs) from precursor RNA of conserved structure that are implicated in translational control (Hutvagner et al., 2001, Science, 293, 834). The RNAi response also features an endonuclease complex containing a siRNA, commonly referred to as an RNA-induced silencing complex (RISC), which mediates cleavage of single-stranded RNA having sequence homologous to the siRNA. Cleavage of the target RNA takes place in the middle of the region complementary to the guide sequence of the siRNA duplex (Elbashir et al., 2001, Genes Dev., 15, 188). In addition, RNA interference can also involve small RNA (e.g., micro-RNA or miRNA) mediated gene silencing, presumably though cellular mechanisms that regulate chromatin structure and thereby prevent transcription of target gene sequences (see for example Allshire, 2002, Science, 297, 1818-1819; Volpe et al., 2002, Science, 297, 1833-1837; Jenuwein, 2002, Science, 297, 2215-2218; and Hall et al., 2002, Science, 297, 2232-2237). As such, siNA molecules of the invention can be used to mediate gene silencing via interaction with RNA transcripts or alternately by interaction with particular gene sequences, wherein such interaction results in gene silencing either at the transcriptional level or post-transcriptional level.

RNAi has been studied in a variety of systems. Fire et al., 1998, Nature, 391, 806, were the first to observe RNAi in C. elegans. Wianny and Goetz, 1999, Nature Cell Biol., 2, 70, describe RNAi mediated by dsRNA in mouse embryos. Hammond et al., 2000, Nature, 404, 293, describe RNAi in Drosophila cells transfected with dsRNA. Elbashir et al., 2001, Nature, 411, 494, describe RNAi induced by introduction of duplexes of synthetic 21-nucleotide RNAs in cultured mammalian cells including human embryonic kidney and HeLa cells. Recent work in Drosophila embryonic lysates has revealed certain requirements for siRNA length, structure, chemical composition, and sequence that are essential to mediate efficient RNAi activity. These studies have shown that 21 nucleotide siRNA duplexes are most active when containing two 2-nucleotide 3′-terminal nucleotide overhangs. Furthermore, substitution of one or both siRNA strands with 2′-deoxy or 2′-O-methyl nucleotides abolishes RNAi activity, whereas substitution of 3′-terminal siRNA nucleotides with deoxy nucleotides was shown to be tolerated. Mismatch sequences in the center of the siRNA duplex were also shown to abolish RNAi activity. In addition, these studies also indicate that the position of the cleavage site in the target RNA is defined by the 5′-end of the siRNA guide sequence rather than the 3′-end (Elbashir et al., 2001, EMBO J., 20, 6877). Other studies have indicated that a 5′-phosphate on the target-complementary strand of a siRNA duplex is required for siRNA activity and that ATP is utilized to maintain the 5′-phosphate moiety on the siRNA (Nykanen et al., 2001, Cell, 107, 309); however, siRNA molecules lacking a 5′-phosphate are active when introduced exogenously, suggesting that 5′-phosphorylation of siRNA constructs may occur in vivo.

Duplex Forming Oligonucleotides (DFO) of the Invention

In one embodiment, the invention features siNA molecules comprising duplex forming oligonucleotides (DFO) that can self-assemble into double stranded oligonucleotides. The duplex forming oligonucleotides of the invention can be chemically synthesized or expressed from transcription units and/or vectors. The DFO molecules of the instant invention provide useful reagents and methods for a variety of therapeutic, diagnostic, agricultural, veterinary, target validation, genomic discovery, genetic engineering and pharmacogenomic applications.

Applicant demonstrates herein that certain oligonucleotides, refered to herein for convenience but not limitation as duplex forming oligonucleotides or DFO molecules, are potent mediators of sequence specific regulation of gene expression. The oligonucleotides of the invention are distinct from other nucleic acid sequences known in the art (e.g., siRNA, miRNA, stRNA, shRNA, antisense oligonucleotides etc.) in that they represent a class of linear polynucleotide sequences that are designed to self-assemble into double stranded oligonucleotides, where each strand in the double stranded oligonucleotides comprises a nucleotide sequence that is complementary to a target nucleic acid molecule. Nucleic acid molecules of the invention can thus self assemble into functional duplexes in which each strand of the duplex comprises the same polynucleotide sequence and each strand comprises a nucleotide sequence that is complementary to a target nucleic acid molecule.

Generally, double stranded oligonucleotides are formed by the assembly of two distinct oligonucleotide sequences where the oligonucleotide sequence of one strand is complementary to the oligonucleotide sequence of the second strand; such double stranded oligonucleotides are assembled from two separate oligonucleotides, or from a single molecule that folds on itself to form a double stranded structure, often referred to in the field as hairpin stem-loop structure (e.g., shRNA or short hairpin RNA). These double stranded oligonucleotides known in the art all have a common feature in that each strand of the duplex has a distict nucleotide sequence.

Distinct from the double stranded nucleic acid molecules known in the art, the applicants have developed a novel, potentially cost effective and simplified method of forming a double stranded nucleic acid molecule starting from a single stranded or linear oligonucleotide. The two strands of the double stranded oligonucleotide formed according to the instant invention have the same nucleotide sequence and are not covalently linked to each other. Such double-stranded oligonucleotides molecules can be readily linked post-synthetically by methods and reagents known in the art and are within the scope of the invention. In one embodiment, the single stranded oligonucleotide of the invention (the duplex forming oligonucleotide) that forms a double stranded oligonucleotide comprises a first region and a second region, where the second region includes a nucleotide sequence that is an inverted repeat of the nucleotide sequence in the first region, or a portion thereof, such that the single stranded oligonucleotide self assembles to form a duplex oligonucleotide in which the nucleotide sequence of one strand of the duplex is the same as the nucleotide sequence of the second strand. Non-limiting examples of such duplex forming oligonucleotides are illustrated in FIGS. 14 and 15. These duplex forming oligonucleotides (DFOs) can optionally include certain palindrome or repeat sequences where such palindrome or repeat sequences are present in between the first region and the second region of the DFO.

In one embodiment, the invention features a duplex forming oligonucleotide (DFO) molecule, wherein the DFO comprises a duplex forming self complementary nucleic acid sequence that has nucleotide sequence complementary to a target nucleic acid sequence. The DFO molecule can comprise a single self complementary sequence or a duplex resulting from assembly of such self complementary sequences.

In one embodiment, a duplex forming oligonucleotide (DFO) of the invention comprises a first region and a second region, wherein the second region comprises a nucleotide sequence comprising an inverted repeat of nucleotide sequence of the first region such that the DFO molecule can assemble into a double stranded oligonucleotide. Such double stranded oligonucleotides can act as a short interfering nucleic acid (siNA) to modulate gene expression. Each strand of the double stranded oligonucleotide duplex formed by DFO molecules of the invention can comprise a nucleotide sequence region that is complementary to the same nucleotide sequence in a target nucleic acid molecule (e.g., target target RNA).

In one embodiment, the invention features a single stranded DFO that can assemble into a double stranded oligonucleotide. The applicant has surprisingly found that a single stranded oligonucleotide with nucleotide regions of self complementarity can readily assemble into duplex oligonucleotide constructs. Such DFOs can assemble into duplexes that can inhibit gene expression in a sequence specific manner. The DFO moleucles of the invention comprise a first region with nucleotide sequence that is complementary to the nucleotide sequence of a second region and where the sequence of the first region is complementary to a target nucleic acid (e.g., RNA). The DFO can form a double stranded oligonucleotide wherein a portion of each strand of the double stranded oligonucleotide comprises a sequence complementary to a target nucleic acid sequence.

In one embodiment, the invention features a double stranded oligonucleotide, wherein the two strands of the double stranded oligonucleotide are not covalently linked to each other, and wherein each strand of the double stranded oligonucleotide comprises a nucleotide sequence that is complementary to the same nucleotide sequence in a target nucleic acid molecule or a portion thereof (e.g., target RNA target). In another embodiment, the two strands of the double stranded oligonucleotide share an identical nucleotide sequence of at least about 15, preferably at least about 16, 17, 18, 19, 20, or 21 nucleotides.

In one embodiment, a DFO molecule of the invention comprises a structure having Formula DFO-I:

5′-p-XZX′-3′

wherein Z comprises a palindromic or repeat nucleic acid sequence optionally with one or more modified nucleotides (e.g., nucleotide with a modified base, such as 2-amino purine, 2-amino-1,6-dihydro purine or a universal base), for example of length about 2 to about 24 nucleotides in even numbers (e.g., about 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, or 22 or 24 nucleotides), X represents a nucleic acid sequence, for example of length of about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 nucleotides), X′ comprises a nucleic acid sequence, for example of length about 1 and about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides) having nucleotide sequence complementarity to sequence X or a portion thereof, p comprises a terminal phosphate group that can be present or absent, and wherein sequence X and Z, either independently or together, comprise nucleotide sequence that is complementary to a target nucleic acid sequence or a portion thereof and is of length sufficient to interact (e.g., base pair) with the target nucleic acid sequence or a portion thereof (e.g., target RNA target). For example, X independently can comprise a sequence from about 12 to about 21 or more (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more) nucleotides in length that is complementary to nucleotide sequence in a target target RNA or a portion thereof. In another non-limiting example, the length of the nucleotide sequence of X and Z together, when X is present, that is complementary to the target RNA or a portion thereof (e.g., target RNA target) is from about 12 to about 21 or more nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more). In yet another non-limiting example, when X is absent, the length of the nucleotide sequence of Z that is complementary to the target target RNA or a portion thereof is from about 12 to about 24 or more nucleotides (e.g., about 12, 14, 16, 18, 20, 22, 24, or more). In one embodiment X, Z and X′ are independently oligonucleotides, where X and/or Z comprises a nucleotide sequence of length sufficient to interact (e.g., base pair) with a nucleotide sequence in the target RNA or a portion thereof (e.g., target RNA target). In one embodiment, the lengths of oligonucleotides X and X′ are identical. In another embodiment, the lengths of oligonucleotides X and X′ are not identical. In another embodiment, the lengths of oligonucleotides X and Z, or Z and X′, or X, Z and X′ are either identical or different.

When a sequence is described in this specification as being of “sufficient” length to interact (i.e., base pair) with another sequence, it is meant that the the length is such that the number of bonds (e.g., hydrogen bonds) formed between the two sequences is enough to enable the two sequence to form a duplex under the conditions of interest. Such conditions can be in vitro (e.g., for diagnostic or assay purposes) or in vivo (e.g., for therapeutic purposes). It is a simple and routine matter to determine such lengths.

In one embodiment, the invention features a double stranded oligonucleotide construct having Formula DFO-I(a):

5′-p-XZX′-3′
3′-X′ZX-p-5′

wherein Z comprises a palindromic or repeat nucleic acid sequence or palindromic or repeat-like nucleic acid sequence with one or more modified nucleotides (e.g., nucleotides with a modified base, such as 2-amino purine, 2-amino-1,6-dihydro purine or a universal base), for example of length about 2 to about 24 nucleotides in even numbers (e.g., about 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 or 24 nucleotides), X represents a nucleic acid sequence, for example of length about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 nucleotides), X′ comprises a nucleic acid sequence, for example of length about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides) having nucleotide sequence complementarity to sequence X or a portion thereof, p comprises a terminal phosphate group that can be present or absent, and wherein each X and Z independently comprises a nucleotide sequence that is complementary to a target nucleic acid sequence or a portion thereof (e.g., target RNA target) and is of length sufficient to interact with the target nucleic acid sequence of a portion thereof (e.g., target RNA target). For example, sequence X independently can comprise a sequence from about 12 to about 21 or more nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more) in length that is complementary to a nucleotide sequence in a target RNA or a portion thereof (e.g., target RNA target). In another non-limiting example, the length of the nucleotide sequence of X and Z together (when X is present) that is complementary to the target target RNA or a portion thereof is from about 12 to about 21 or more nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more). In yet another non-limiting example, when X is absent, the length of the nucleotide sequence of Z that is complementary to the target target RNA or a portion thereof is from about 12 to about 24 or more nucleotides (e.g., about 12, 14, 16, 18, 20, 22, 24 or more). In one embodiment X, Z and X′ are independently oligonucleotides, where X and/or Z comprises a nucleotide sequence of length sufficient to interact (e.g., base pair) with nucleotide sequence in the target RNA or a portion thereof (e.g., target RNA target). In one embodiment, the lengths of oligonucleotides X and X′ are identical. In another embodiment, the lengths of oligonucleotides X and X′ are not identical. In another embodiment, the lengths of oligonucleotides X and Z or Z and X′ or X, Z and X′ are either identical or different. In one embodiment, the double stranded oligonucleotide construct of Formula I(a) includes one or more, specifically 1, 2, 3 or 4, mismatches, to the extent such mismatches do not significantly diminish the ability of the double stranded oligonucleotide to inhibit target gene expression.

In one embodiment, a DFO molecule of the invention comprises structure having Formula DFO-II:

5′-p-XX′-3′

wherein each X and X′ are independently oligonucleotides of length about 12 nucleotides to about 21 nucleotides, wherein X comprises, for example, a nucleic acid sequence of length about 12 to about 21 nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides), X′ comprises a nucleic acid sequence, for example of length about 12 to about 21 nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 nucleotides) having nucleotide sequence complementarity to sequence X or a portion thereof, p comprises a terminal phosphate group that can be present or absent, and wherein X comprises a nucleotide sequence that is complementary to a target nucleic acid sequence (e.g., target RNA) or a portion thereof and is of length sufficient to interact (e.g., base pair) with the target nucleic acid sequence of a portion thereof. In one embodiment, the length of oligonucleotides X and X′ are identical. In another embodiment the length of oligonucleotides X and X′ are not identical. In one embodiment, length of the oligonucleotides X and X′ are sufficint to form a relatively stable double stranded oligonucleotide.

In one embodiment, the invention features a double stranded oligonucleotide construct having Formula DFO-II(a):

5′-p-XX′-3′
3′-X′X-p-5′

wherein each X and X′ are independently oligonucleotides of length about 12 nucleotides to about 21 nucleotides, wherein X comprises a nucleic acid sequence, for example of length about 12 to about 21 nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides), X′ comprises a nucleic acid sequence, for example of length about 12 to about 21 nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides) having nucleotide sequence complementarity to sequence X or a portion thereof, p comprises a terminal phosphate group that can be present or absent, and wherein X comprises nucleotide sequence that is complementary to a target nucleic acid sequence or a portion thereof (e.g., target RNA target) and is of length sufficient to interact (e.g., base pair) with the target nucleic acid sequence (e.g., target RNA) or a portion thereof. In one embodiment, the lengths of oligonucleotides X and X′ are identical. In another embodiment, the lengths of oligonucleotides X and X′ are not identical. In one embodiment, the lengths of the oligonucleotides X and X′ are sufficint to form a relatively stable double stranded oligonucleotide. In one embodiment, the double stranded oligonucleotide construct of Formula II(a) includes one or more, specifically 1, 2, 3 or 4, mismatches, to the extent such mismatches do not significantly diminish the ability of the double stranded oligonucleotide to inhibit target gene expression.

In one embodiment, the invention features a DFO molecule having Formula DFO-I(b):

5′-p-Z-3′

where Z comprises a palindromic or repeat nucleic acid sequence optionally including one or more non-standard or modified nucleotides (e.g., nucleotide with a modified base, such as 2-amino purine or a universal base) that can facilitate base-pairing with other nucleotides. Z can be, for example, of length sufficient to interact (e.g., base pair) with nucleotide sequence of a target nucleic acid (e.g., target RNA) molecule, preferably of length of at least 12 nucleotides, specifically about 12 to about 24 nucleotides (e.g., about 12, 14, 16, 18, 20, 22 or 24 nucleotides). p represents a terminal phosphate group that can be present or absent.

In one embodiment, a DFO molecule having any of Formula DFO-I, DFO-I(a), DFO-I(b), DFO-II(a) or DFO-II can comprise chemical modifications as described herein without limitation, such as, for example, nucleotides having any of Formulae I-VII, stabilization chemistries as described in Table IV, or any other combination of modified nucleotides and non-nucleotides as described in the various embodiments herein.

In one embodiment, the palidrome or repeat sequence or modified nucleotide (e.g., nucleotide with a modified base, such as 2-amino purine or a universal base) in Z of DFO constructs having Formula DFO-I, DFO-I(a) and DFO-I(b), comprises chemically modified nucleotides that are able to interact with a portion of the target nucleic acid sequence (e.g., modified base analogs that can form Watson Crick base pairs or non-Watson Crick base pairs).

In one embodiment, a DFO molecule of the invention, for example a DFO having Formula DFO-I or DFO-II, comprises about 15 to about 40 nucleotides (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 nucleotides). In one embodiment, a DFO molecule of the invention comprises one or more chemical modifications. In a non-limiting example, the introduction of chemically modified nucleotides and/or non-nucleotides into nucleic acid molecules of the invention provides a powerful tool in overcoming potential limitations of in vivo stability and bioavailability inherent to unmodified RNA molecules that are delivered exogenously. For example, the use of chemically modified nucleic acid molecules can enable a lower dose of a particular nucleic acid molecule for a given therapeutic effect since chemically modified nucleic acid molecules tend to have a longer half-life in serum or in cells or tissues. Furthermore, certain chemical modifications can improve the bioavailability and/or potency of nucleic acid molecules by not only enhancing half-life but also facilitating the targeting of nucleic acid molecules to particular organs, cells or tissues and/or improving cellular uptake of the nucleic acid molecules. Therefore, even if the activity of a chemically modified nucleic acid molecule is reduced in vitro as compared to a native/unmodified nucleic acid molecule, for example when compared to an unmodified RNA molecule, the overall activity of the modified nucleic acid molecule can be greater than the native or unmodified nucleic acid molecule due to improved stability, potency, duration of effect, bioavailability and/or delivery of the molecule.

Multifunctional or Multi-Targeted siNA Molecules of the Invention

In one embodiment, the invention features siNA molecules comprising multifunctional short interfering nucleic acid (multifunctional siNA) molecules that modulate the expression of one or more genes in a biologic system, such as a cell, tissue, or organism. The multifunctional short interfering nucleic acid (multifunctional siNA) molecules of the invention can target more than one region a target nucleic acid sequence or can target sequences of more than one distinct target nucleic acid molecules. The multifunctional siNA molecules of the invention can be chemically synthesized or expressed from transcription units and/or vectors. The multifunctional siNA molecules of the instant invention provide useful reagents and methods for a variety of human applications, therapeutic, cosmetic, diagnostic, agricultural, veterinary, target validation, genomic discovery, genetic engineering and pharmacogenomic applications.

Applicant demonstrates herein that certain oligonucleotides, refered to herein for convenience but not limitation as multifunctional short interfering nucleic acid or multifunctional siNA molecules, are potent mediators of sequence specific regulation of gene expression. The multifunctional siNA molecules of the invention are distinct from other nucleic acid sequences known in the art (e.g., siRNA, miRNA, stRNA, shRNA, antisense oligonucleotides, etc.) in that they represent a class of polynucleotide molecules that are designed such that each strand in the multifunctional siNA construct comprises a nucleotide sequence that is complementary to a distinct nucleic acid sequence in one or more target nucleic acid molecules. A single multifunctional siNA molecule (generally a double-stranded molecule) of the invention can thus target more than one (e.g., 2, 3, 4, 5, or more) differing target nucleic acid target molecules. Nucleic acid molecules of the invention can also target more than one (e.g., 2, 3, 4, 5, or more) region of the same target nucleic acid sequence. As such multifunctional siNA molecules of the invention are useful in down regulating or inhibiting the expression of one or more target nucleic acid molecules. By reducing or inhibiting expression of more than one target nucleic acid molecule with one multifunctional siNA construct, multifunctional siNA molecules of the invention represent a class of potent therapeutic agents that can provide simultaneous inhibition of multiple targets within a disease or pathogen related pathway. Such simultaneous inhibition can provide synergistic therapeutic treatment strategies without the need for separate preclinical and clinical development efforts or complex regulatory approval process.

Use of multifunctional siNA molecules that target more then one region of a target nucleic acid molecule (e.g., messenger RNA) is expected to provide potent inhibition of gene expression. For example, a single multifunctional siNA construct of the invention can target both conserved and variable regions of a target nucleic acid molecule, such as a target RNA or DNA, thereby allowing down regulation or inhibition of different splice variants encoded by a single gene, or allowing for targeting of both coding and non-coding regions of a target nucleic acid molecule.

Generally, double stranded oligonucleotides are formed by the assembly of two distinct oligonucleotides where the oligonucleotide sequence of one strand is complementary to the oligonucleotide sequence of the second strand; such double stranded oligonucleotides are generally assembled from two separate oligonucleotides (e.g., siRNA). Alternately, a duplex can be formed from a single molecule that folds on itself (e.g., shRNA or short hairpin RNA). These double stranded oligonucleotides are known in the art to mediate RNA interference and all have a common feature wherein only one nucleotide sequence region (guide sequence or the antisense sequence) has complementarity to a target nucleic acid sequence, and the other strand (sense sequence) comprises nucleotide sequence that is homologous to the target nucleic acid sequence. Generally, the antisense sequence is retained in the active RISC complex and guides the RISC to the target nucleotide sequence by means of complementary base-pairing of the antisense sequence with the target seqeunce for mediating sequence-specific RNA interference. It is known in the art that in some cell culture systems, certain types of unmodified siRNAs can exhibit “off target” effects. It is hypothesized that this off-target effect involves the participation of the sense sequence instead of the antisense sequence of the siRNA in the RISC complex (see for example Schwarz et al., 2003, Cell, 115, 199-208). In this instance the sense sequence is believed to direct the RISC complex to a sequence (off-target sequence) that is distinct from the intended target sequence, resulting in the inhibition of the off-target sequence. In these double stranded nucleic acid molecules, each strand is complementary to a distinct target nucleic acid sequence. However, the off-targets that are affected by these dsRNAs are not entirely predictable and are non-specific.

Distinct from the double stranded nucleic acid molecules known in the art, the applicants have developed a novel, potentially cost effective and simplified method of down regulating or inhibiting the expression of more than one target nucleic acid sequence using a single multifunctional siNA construct. The multifunctional siNA molecules of the invention are designed to be double-stranded or partially double stranded, such that a portion of each strand or region of the multifunctional siNA is complementary to a target nucleic acid sequence of choice. As such, the multifunctional siNA molecules of the invention are not limited to targeting sequences that are complementary to each other, but rather to any two differing target nucleic acid sequences. Multifunctional siNA molecules of the invention are designed such that each strand or region of the multifunctional siNA molecule, that is complementary to a given target nucleic acid sequence, is of suitable length (e.g., from about 16 to about 28 nucleotides in length, preferably from about 18 to about 28 nucleotides in length) for mediating RNA interference against the target nucleic acid sequence. The complementarity between the target nucleic acid sequence and a strand or region of the multifunctional siNA must be sufficient (at least about 8 base pairs) for cleavage of the target nucleic acid sequence by RNA interference. multifunctional siNA of the invention is expected to minimize off-target effects seen with certain siRNA sequences, such as those described in (Schwarz et al., supra).

It has been reported that dsRNAs of length between 29 base pairs and 36 base pairs (Tuschl et al., International PCT Publication No. WO 02/44321) do not mediate RNAi. One reason these dsRNAs are inactive may be the lack of turnover or dissociation of the strand that interacts with the target RNA sequence, such that the RISC complex is not able to efficiently interact with multiple copies of the target RNA resulting in a significant decrease in the potency and efficiency of the RNAi process. Applicant has surprisingly found that the multifunctional siNAs of the invention can overcome this hurdle and are capable of enhancing the efficiency and potency of RNAi process. As such, in certain embodiments of the invention, multifunctional siNAs of length of about 29 to about 36 base pairs can be designed such that, a portion of each strand of the multifunctional siNA molecule comprises a nucleotide sequence region that is complementary to a target nucleic acid of length sufficient to mediate RNAi efficiently (e.g., about 15 to about 23 base pairs) and a nucleotide sequence region that is not complementary to the target nucleic acid. By having both complementary and non-complementary portions in each strand of the multifunctional siNA, the multifunctional siNA can mediate RNA interference against a target nucleic acid sequence without being prohibitive to turnover or dissociation (e.g., where the length of each strand is too long to mediate RNAi against the respective target nucleic acid sequence). Furthermore, design of multifunctional siNA molecules of the invention with internal overlapping regions allows the multifunctional siNA molecules to be of favorable (decreased) size for mediating RNA interference and of size that is well suited for use as a therapeutic agent (e.g., wherein each strand is independently from about 18 to about 28 nucleotides in length). Non-limiting examples are illustrated in FIGS. 16-28.

In one embodiment, a multifunctional siNA molecule of the invention comprises a first region and a second region, where the first region of the multifunctional siNA comprises a nucleotide sequence complementary to a nucleic acid sequence of a first target nucleic acid molecule, and the second region of the multifunctional siNA comprises nucleic acid sequence complementary to a nucleic acid sequence of a second target nucleic acid molecule. In one embodiment, a multifunctional siNA molecule of the invention comprises a first region and a second region, where the first region of the multifunctional siNA comprises nucleotide sequence complementary to a nucleic acid sequence of the first region of a target nucleic acid molecule, and the second region of the multifunctional siNA comprises nucleotide sequence complementary to a nucleic acid sequence of a second region of a the target nucleic acid molecule. In another embodiment, the first region and second region of the multifunctional siNA can comprise separate nucleic acid sequences that share some degree of complementarity (e.g., from about 1 to about 10 complementary nucleotides). In certain embodiments, multifunctional siNA constructs comprising separate nucleic acid seqeunces can be readily linked post-synthetically by methods and reagents known in the art and such linked constructs are within the scope of the invention. Alternately, the first region and second region of the multifunctional siNA can comprise a single nucleic acid sequence having some degree of self complementarity, such as in a hairpin or stem-loop structure. Non-limiting examples of such double stranded and hairpin multifunctional short interfering nucleic acids are illustrated in FIGS. 16 and 17 respectively. These multifunctional short interfering nucleic acids (multifunctional siNAs) can optionally include certain overlapping nucleotide sequence where such overlapping nucleotide sequence is present in between the first region and the second region of the multifunctional siNA (see for example FIGS. 18 and 19). In one embodiment, the first target nucleic acid molecule and the second nucleic acid target molecule are one or more HDCA target sequences, such as any HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7 nucleic acid sequence.

In one embodiment, the invention features a multifunctional short interfering nucleic acid (multifunctional siNA) molecule, wherein each strand of the the multifunctional siNA independently comprises a first region of nucleic acid sequence that is complementary to a distinct target nucleic acid sequence and the second region of nucleotide sequence that is not complementary to the target sequence. The target nucleic acid sequence of each strand is in the same target nucleic acid molecule or different target nucleic acid molecules. In one embodiment, the nucleic acid target molecule(s) comprises one or more HDCA target sequence, such as any HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7 nucleic acid sequence.

In another embodiment, the multifunctional siNA comprises two strands, where: (a) the first strand comprises a region having sequence complementarity to a target nucleic acid sequence (complementary region 1) and a region having no sequence complementarity to the target nucleotide sequence (non-complementary region 1); (b) the second strand of the multifunction siNA comprises a region having sequence complementarity to a target nucleic acid sequence that is distinct from the target nucleotide sequence complementary to the first strand nucleotide sequence (complementary region 2), and a region having no sequence complementarity to the target nucleotide sequence of complementary region 2 (non-complementary region 2); (c) the complementary region 1 of the first strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in the non-complementary region 2 of the second strand and the complementary region 2 of the second strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in the non-complementary region 1 of the first strand. The target nucleic acid sequence of complementary region 1 and complementary region 2 is in the same target nucleic acid molecule or different target nucleic acid molecules. In one embodiment, the nucleic acid target molecule(s) comprises one or more HDCA target sequence, such as any HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7 nucleic acid sequence.

In another embodiment, the multifunctional siNA comprises two strands, where: (a) the first strand comprises a region having sequence complementarity to a target nucleic acid sequence derived from a gene (complementary region 1) and a region having no sequence complementarity to the target nucleotide sequence of complementary region 1 (non-complementary region 1); (b) the second strand of the multifunction siNA comprises a region having sequence complementarity to a target nucleic acid sequence derived from a gene that is distinct from the gene of complementary region 1 (complementary region 2), and a region having no sequence complementarity to the target nucleotide sequence of complementary region 2 (non-complementary region 2); (c) the complementary region 1 of the first strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in the non-complementary region 2 of the second strand and the complementary region 2 of the second strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in the non-complementary region 1 of the first strand. In one embodiment, the nucleic acid target sequence comprises one or more HDCA target sequences, such as any HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7 nucleic acid sequence.

In another embodiment, the multifunctional siNA comprises two strands, where: (a) the first strand comprises a region having sequence complementarity to a target nucleic acid sequence derived from a first gene (complementary region 1) and a region having no sequence complementarity to the target nucleotide sequence of complementary region 1 (non-complementary region 1); (b) the second strand of the multifunction siNA comprises a region having sequence complementarity to a second target nucleic acid sequence distinct from the first target nucleic acid sequence of complementary region 1 (complementary region 2), provided, however, that the target nucleic acid sequence for complementary region 1 and target nucleic acid sequence for complementary region 2 are both derived from the same gene, and a region having no sequence complementarity to the target nucleotide sequence of complementary region 2 (non-complementary region 2); (c) the complementary region 1 of the first strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in the non-complementary region 2 of the second strand and the complementary region 2 of the second strand comprises a nucleotide sequence that is complementary to nucleotide sequence in the non-complementary region 1 of the first strand. In one embodiment, the nucleic acid target sequence comprises one or more HDCA target sequences, such as any HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7 nucleic acid sequence.

In one embodiment, the invention features a multifunctional short interfering nucleic acid (multifunctional siNA) molecule, wherein the multifunctional siNA comprises two complementary nucleic acid sequences in which the first sequence comprises a first region having nucleotide sequence complementary to nucleotide sequence within a first target nucleic acid molecule, and in which the second seqeunce comprises a first region having nucleotide sequence complementary to a distinct nucleotide sequence within the same target nucleic acid molecule. Preferably, the first region of the first sequence is also complementary to the nucleotide sequence of the second region of the second sequence, and where the first region of the second sequence is complementary to the nucleotide sequence of the second region of the first sequence. In one embodiment, the nucleic acid target sequence comprises one or more HDCA target sequences, such as any HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7 nucleic acid sequence.

In one embodiment, the invention features a multifunctional short interfering nucleic acid (multifunctional siNA) molecule, wherein the multifunctional siNA comprises two complementary nucleic acid sequences in which the first sequence comprises a first region having a nucleotide sequence complementary to a nucleotide sequence within a first target nucleic acid molecule, and in which the second seqeunce comprises a first region having a nucleotide sequence complementary to a distinct nucleotide sequence within a second target nucleic acid molecule. Preferably, the first region of the first sequence is also complementary to the nucleotide sequence of the second region of the second sequence, and where the first region of the second sequence is complementary to the nucleotide sequence of the second region of the first sequence. In one embodiment, the nucleic acid target sequence comprises one or more HDCA target sequences, such as any HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7 nucleic acid sequence.

In one embodiment, the invention features a multifunctional siNA molecule comprising a first region and a second region, where the first region comprises a nucleic acid sequence having about 18 to about 28 nucleotides complementary to a nucleic acid sequence within a first target nucleic acid molecule, and the second region comprises nucleotide sequence having about 18 to about 28 nucleotides complementary to a distinct nucleic acid sequence within a second target nucleic acid molecule. In one embodiment, the first nucleic acid target molecule and the second target nucleic acid molecule are selected from the group consisting of any of HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7 nucleic acid sequences.

In one embodiment, the invention features a multifunctional siNA molecule comprising a first region and a second region, where the first region comprises nucleic acid sequence having about 18 to about 28 nucleotides complementary to a nucleic acid sequence within a target nucleic acid molecule, and the second region comprises nucleotide sequence having about 18 to about 28 nucleotides complementary to a distinct nucleic acid sequence within the same target nucleic acid molecule. In one embodiment, the nucleic acid target molecule is selected from the group consisting of any of HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7 nucleic acid sequences.

In one embodiment, the invention features a double stranded multifunctional short interfering nucleic acid (multifunctional siNA) molecule, wherein one strand of the multifunctional siNA comprises a first region having nucleotide sequence complementary to a first target nucleic acid sequence, and the second strand comprises a first region having a nucleotide sequence complementary to a second target nucleic acid sequence. The first and second target nucleic acid sequences can be present in separate target nucleic acid molecules or can be different regions within the same target nucleic acid molecule. As such, multifunctional siNA molecules of the invention can be used to target the expression of different genes, splice variants of the same gene, both mutant and conserved regions of one or more gene transcripts, or both coding and non-coding sequences of the same or differeing genes or gene transcripts. In one embodiment, the first nucleic acid target sequence and the second target nucleic acid sequence are selected from the group consisting of any of HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7 nucleic acid sequences.

In one embodiment, a target nucleic acid molecule of the invention encodes a single protein. In another embodiment, a target nucleic acid molecule encodes more than one protein (e.g., 1, 2, 3, 4, 5 or more proteins). As such, a multifunctional siNA construct of the invention can be used to down regulate or inhibit the expression of several proteins (e.g., any of HDAC 1, HDAC 2, HCAC 3, HDAC 4, HDAC 5, HDAC 6, HDAC 7, HDAC 8, HDAC 9a, HDAC 9b, HDAC 10, and/or HDAC 11, and/or SIR T1, 2, 3, 4, 5, 6, and/or 7 proteins). For example, a multifunctional siNA molecule comprising a region in one strand having nucleotide sequence complementarity to a first target nucleic acid sequence derived from a gene encoding one protein and the second strand comprising a region with nucleotide sequence complementarity to a second target nucleic acid sequence present in target nucleic acid molecules derived from genes encoding two or more proteins (e.g., two or more differing target sequences) can be used to down regulate, inhibit, or shut down a particular biologic pathway by targeting, for example, two or more targets involved in a biologic pathway.

In one embodiment the invention takes advantage of conserved nucleotide sequences present in different isoforms of cytokines or ligands and receptors for the cytokines or ligands. By designing multifunctional siNAs in a manner where one strand includes a sequence that is complementary to a target nucleic acid sequence conserved among various isoforms of a cytokine and the other strand includes sequence that is complementary to a target nucleic acid sequence conserved among the receptors for the cytokine, it is possible to selectively and effectively modulate or inhibit a biological pathway or multiple genes in a biological pathway using a single multifunctional siNA.

In one embodiment, a double stranded multifunctional siNA molecule of the invention comprises a structure having Formula MF-I:

5′-p-XZX′-3′
3′-Y′ZY-p-5′

wherein each 5′-p-XZX′-3′ and 5′-p-YZY′-3′ are independently an oligonucleotide of length of about 20 nucleotides to about 300 nucleotides, preferably of about 20 to about 200 nucleotides, about 20 to about 100 nucleotides, about 20 to about 40 nucleotides, about 20 to about 40 nucleotides, about 24 to about 38 nucleotides, or about 26 to about 38 nucleotides; XZ comprises a nucleic acid sequence that is complementary to a first target nucleic acid sequence; YZ is an oligonucleotide comprising nucleic acid sequence that is complementary to a second target nucleic acid sequence; Z comprises nucleotide sequence of length about 1 to about 24 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 nucleotides) that is self complimentary; X comprises nucleotide sequence of length about 1 to about 100 nucleotides, preferably about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 nucleotides) that is complementary to nucleotide sequence present in region Y′; Y comprises nucleotide sequence of length about 1 to about 100 nucleotides, prefereably about 1-about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides) that is complementary to nucleotide sequence present in region X′; each p comprises a terminal phosphate group that is independently present or absent; each XZ and YZ is independently of length sufficient to stably interact (i.e., base pair) with the first and second target nucleic acid sequence, respectively, or a portion thereof. For example, each sequence X and Y can independently comprise sequence from about 12 to about 21 or more nucleotides in length (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more) that is complementary to a target nucleotide sequence in different target nucleic acid molecules, such as target RNAs or a portion thereof. In another non-limiting example, the length of the nucleotide sequence of X and Z together that is complementary to the first target nucleic acid sequence or a portion thereof is from about 12 to about 21 or more nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more). In another non-limiting example, the length of the nucleotide sequence of Y and Z together, that is complementary to the second target nucleic acid sequence or a portion thereof is from about 12 to about 21 or more nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more). In one embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in the same target nucleic acid molecule (e.g., target RNA). In another embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in different target nucleic acid molecules. In one embodiment, Z comprises a palindrome or a repeat sequence. In one embodiment, the lengths of oligonucleotides X and X′ are identical. In another embodiment, the lengths of oligonucleotides X and X′ are not identical. In one embodiment, the lengths of oligonucleotides Y and Y′ are identical. In another embodiment, the lengths of oligonucleotides Y and Y′ are not identical. In one embodiment, the double stranded oligonucleotide construct of Formula I(a) includes one or more, specifically 1, 2, 3 or 4, mismatches, to the extent such mismatches do not significantly diminish the ability of the double stranded oligonucleotide to inhibit target gene expression.

In one embodiment, a multifunctional siNA molecule of the invention comprises a structure having Formula MF-II:

5′-p-XX′-3′
3′-Y′Y-p-5′

wherein each 5′-p-XX′-3′ and 5′-p-YY′-3′ are independently an oligonucleotide of length of about 20 nucleotides to about 300 nucleotides, preferably about 20 to about 200 nucleotides, about 20 to about 100 nucleotides, about 20 to about 40 nucleotides, about 20 to about 40 nucleotides, about 24 to about 38 nucleotides, or about 26 to about 38 nucleotides; X comprises a nucleic acid sequence that is complementary to a first target nucleic acid sequence; Y is an oligonucleotide comprising nucleic acid sequence that is complementary to a second target nucleic acid sequence; X comprises a nucleotide sequence of length about 1 to about 100 nucleotides, preferably about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 nucleotides) that is complementary to nucleotide sequence present in region Y′; Y comprises nucleotide sequence of length about 1 to about 100 nucleotides, prefereably about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides) that is complementary to nucleotide sequence present in region X′; each p comprises a terminal phosphate group that is independently present or absent; each X and Y independently is of length sufficient to stably interact (i.e., base pair) with the first and second target nucleic acid sequence, respectively, or a portion thereof. For example, each sequence X and Y can independently comprise sequence from about 12 to about 21 or more nucleotides in length (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more) that is complementary to a target nucleotide sequence in different target nucleic acid molecules or a portion thereof. In one embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in the same target nucleic acid molecule (e.g., target RNA or DNA). In another embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in different target nucleic acid molecules or a portion thereof. In one embodiment, Z comprises a palindrome or a repeat sequence. In one embodiment, the lengths of oligonucleotides X and X′ are identical. In another embodiment, the lengths of oligonucleotides X and X′ are not identical. In one embodiment, the lengths of oligonucleotides Y and Y′ are identical. In another embodiment, the lengths of oligonucleotides Y and Y′ are not identical. In one embodiment, the double stranded oligonucleotide construct of Formula I(a) includes one or more, specifically 1, 2, 3 or 4, mismatches, to the extent such mismatches do not significantly diminish the ability of the double stranded oligonucleotide to inhibit target gene expression.

In one embodiment, a multifunctional siNA molecule of the invention comprises a structure having Formula MF-III:

XX′
Y′—W—Y

wherein each X, X′, Y, and Y′ is independently an oligonucleotide of length of about 15 nucleotides to about 50 nucleotides, preferably about 18 to about 40 nucleotides, or about 19 to about 23 nucleotides; X comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y′; X′ comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y; each X and X′ is independently of length sufficient to stably interact (i.e., base pair) with a first and a second target nucleic acid sequence, respectively, or a portion thereof; W represents a nucleotide or non-nucleotide linker that connects sequences Y′ and Y; and the multifunctional siNA directs cleavage of the first and second target sequence via RNA interference. In one embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in the same target nucleic acid molecule (e.g., target RNA). In another embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in different target nucleic acid molecules or a portion thereof. In one embodiment, region W connects the 3′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, region W connects the 3′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X′. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y′. In one embodiment, W connects sequences Y and Y′ via a biodegradable linker. In one embodiment, W further comprises a conjugate, label, aptamer, ligand, lipid, or polymer.

In one embodiment, a multifunctional siNA molecule of the invention comprises a structure having Formula MF-IV:

XX′
Y′—W—Y

wherein each X, X′, Y, and Y′ is independently an oligonucleotide of length of about 15 nucleotides to about 50 nucleotides, preferably about 18 to about 40 nucleotides, or about 19 to about 23 nucleotides; X comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y′; X′ comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y; each Y and Y′ is independently of length sufficient to stably interact (i.e., base pair) with a first and a second target nucleic acid sequence, respectively, or a portion thereof; W represents a nucleotide or non-nucleotide linker that connects sequences Y′ and Y; and the multifunctional siNA directs cleavage of the first and second target sequence via RNA interference. In one embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in the same target nucleic acid molecule (e.g., target RNA). In another embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in different target nucleic acid molecules or a portion thereof. In one embodiment, region W connects the 3′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, region W connects the 3′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X′. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y′. In one embodiment, W connects sequences Y and Y′ via a biodegradable linker. In one embodiment, W further comprises a conjugate, label, aptamer, ligand, lipid, or polymer.

In one embodiment, a multifunctional siNA molecule of the invention comprises a structure having Formula MF-V:

XX′
Y′—W—Y

wherein each X, X′, Y, and Y′ is independently an oligonucleotide of length of about 15 nucleotides to about 50 nucleotides, preferably about 18 to about 40 nucleotides, or about 19 to about 23 nucleotides; X comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y′; X′ comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y; each X, X′, Y, or Y′ is independently of length sufficient to stably interact (i.e., base pair) with a first, second, third, or fourth target nucleic acid sequence, respectively, or a portion thereof; W represents a nucleotide or non-nucleotide linker that connects sequences Y′ and Y; and the multifunctional siNA directs cleavage of the first, second, third, and/or fourth target sequence via RNA interference. In one embodiment, the first, second, third and fourth target nucleic acid sequence are all present in the same target nucleic acid molecule (e.g., target RNA). In another embodiment, the first, second, third and fourth target nucleic acid sequence are independently present in different target nucleic acid molecules or a portion thereof. In one embodiment, region W connects the 3′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, region W connects the 3′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X′. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y′. In one embodiment, W connects sequences Y and Y′ via a biodegradable linker. In one embodiment, W further comprises a conjugate, label, aptamer, ligand, lipid, or polymer.

In one embodiment, regions X and Y of multifunctional siNA molecule of the invention (e.g., having any of Formula MF-I-MF-V), are complementary to different target nucleic acid sequences that are portions of the same target nucleic acid molecule. In one embodiment, such target nucleic acid sequences are at different locations within the coding region of a RNA transcript. In one embodiment, such target nucleic acid sequences comprise coding and non-coding regions of the same RNA transcript. In one embodiment, such target nucleic acid sequences comprise regions of alternately spliced transcripts or precursors of such alternately spliced transcripts.

In one embodiment, a multifunctional siNA molecule having any of Formula MF-I-MF-V can comprise chemical modifications as described herein without limitation, such as, for example, nucleotides having any of Formulae I-VII described herein, stabilization chemistries as described in Table IV, or any other combination of modified nucleotides and non-nucleotides as described in the various embodiments herein.

In one embodiment, the palidrome or repeat sequence or modified nucleotide (e.g., nucleotide with a modified base, such as 2-amino purine or a universal base) in Z of multifunctional siNA constructs having Formula MF-I or MF-II comprises chemically modified nucleotides that are able to interact with a portion of the target nucleic acid sequence (e.g., modified base analogs that can form Watson Crick base pairs or non-Watson Crick base pairs).

In one embodiment, a multifunctional siNA molecule of the invention, for example each strand of a multifunctional siNA having MF-I-MF-V, independently comprises about 15 to about 40 nucleotides (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 nucleotides). In one embodiment, a multifunctional siNA molecule of the invention comprises one or more chemical modifications. In a non-limiting example, the introduction of chemically modified nucleotides and/or non-nucleotides into nucleic acid molecules of the invention provides a powerful tool in overcoming potential limitations of in vivo stability and bioavailability inherent to unmodified RNA molecules that are delivered exogenously. For example, the use of chemically modified nucleic acid molecules can enable a lower dose of a particular nucleic acid molecule for a given therapeutic effect since chemically modified nucleic acid molecules tend to have a longer half-life in serum or in cells or tissues. Furthermore, certain chemical modifications can improve the bioavailability and/or potency of nucleic acid molecules by not only enhancing half-life but also facilitating the targeting of nucleic acid molecules to particular organs, cells or tissues and/or improving cellular uptake of the nucleic acid molecules. Therefore, even if the activity of a chemically modified nucleic acid molecule is reduced in vitro as compared to a native/unmodified nucleic acid molecule, for example when compared to an unmodified RNA molecule, the overall activity of the modified nucleic acid molecule can be greater than the native or unmodified nucleic acid molecule due to improved stability, potency, duration of effect, bioavailability and/or delivery of the molecule.

In another embodiment, the invention features multifunctional siNAs, wherein the multifunctional siNAs are assembled from two separate double-stranded siNAs, where one of the ends of each sense strand is tethered to the end of the sense strand of the other siNA molecule, such that the two antisense siNA strands are annealed to their corresponding sense strand that are tethered to each other at one end (see FIG. 22). The tethers or linkers can be nucleotide-based linkers or non-nucleotide based linkers as generally known in the art and as described herein.

In one embodiment, the invention features a multifunctional siNA, wherein the multifunctional siNA is assembled from two separate double-stranded siNAs, where the 5′-end of one antisense strand of the siNA is tethered to the 3′-end of the antisense strand of the other siNA molecule, such that the 5′-end of the one of the sense siNA strands annealed to their corresponding antisense sense strand that are tethered to each other at one end, faces the 3′-end of the other sense strand (see FIG. 22 (G-H)). In one embodiment, the linkage between the 5′-end of the first antisense strand and the 3′-end of the second antisense strand is designed in such a way as to be readily cleavable (e.g., biodegradable linker) such that the 5′end of each antisense strand of the multifunctional siNA has a free 5′-end suitable to mediate RNA interefence-based cleavage of the target RNA. The tethers or linkers can be nucleotide-based linkers or non-nucleotide based linkers as generally known in the art and as described herein.

In one embodiment, the invention features a multifunctional siNA, wherein the multifunctional siNA is assembled from two separate double-stranded siNAs, where the 5′-end of one antisense strand of the siNA is tethered to the 5′-end of the antisense strand of the other siNA molecule, such that the 3′-end of the one of the sense siNA strands annealed to their corresponding antisense sense strand that are tethered to each other at one end, faces the 3′-end of the other sense strand (see FIG. 22 (E)). In one embodiment, the linkage between the 5′-end of the first antisense strand and the 5′-end of the second antisense strand is designed in such a way as to be readily cleavable (e.g., biodegradable linker) such that the 5′end of each antisense strand of the multifunctional siNA has a free 5′-end suitable to mediate RNA interefence-based cleavage of the target RNA. The tethers or linkers can be nucleotide-based linkers or non-nucleotide based linkers as generally known in the art and as described herein.

In one embodiment, the invention features a multifunctional siNA, wherein the multifunctional siNA is assembled from two separate double-stranded siNAs, where the 3′-end of one antisense strand of the siNA is tethered to the 3′-end of the antisense strand of the other siNA molecule, such that the 5′-end of the one of the sense siNA strands annealed to their corresponding antisense sense strand that are tethered to each other at one end, faces the 3′-end of the other sense strand (see FIG. 22 (F)). In one embodiment, the linkage between the 5′-end of the first antisense strand and the 5′-end of the second antisense strand is designed in such a way as to be readily cleavable (e.g., biodegradable linker) such that the 5′end of each antisense strand of the multifunctional siNA has a free 5′-end suitable to mediate RNA interefence-based cleavage of the target RNA. The tethers or linkers can be nucleotide-based linkers or non-nucleotide based linkers as generally known in the art and as described herein.

In any of the above embodiments, a first target nucleic acid sequence or second target nucleic acid sequence can independently comprise target RNA, DNA or a portion thereof. In one embodiment, the first target nucleic acid sequence is a target RNA, DNA or a portion thereof and the second target nucleic acid sequence is a target RNA, DNA of a portion thereof. In one embodiment, the first target nucleic acid sequence is a target RNA, DNA or a portion thereof and the second target nucleic acid sequence is another RNA, DNA of a portion thereof.

Synthesis of Nucleic Acid Molecules

Synthesis of nucleic acids greater than 100 nucleotides in length is difficult using automated methods, and the therapeutic cost of such molecules is prohibitive. In this invention, small nucleic acid motifs (“small” refers to nucleic acid motifs no more than 100 nucleotides in length, preferably no more than 80 nucleotides in length, and most preferably no more than 50 nucleotides in length; e.g., individual siNA oligonucleotide sequences or siNA sequences synthesized in tandem) are preferably used for exogenous delivery. The simple structure of these molecules increases the ability of the nucleic acid to invade targeted regions of protein and/or RNA structure. Exemplary molecules of the instant invention are chemically synthesized, and others can similarly be synthesized.

Oligonucleotides (e.g., certain modified oligonucleotides or portions of oligonucleotides lacking ribonucleotides) are synthesized using protocols known in the art, for example as described in Caruthers et al., 1992, Methods in Enzymology 211, 3-19, Thompson et al., International PCT Publication No. WO 99/54459, Wincott et al., 1995, Nucleic Acids Res. 23, 2677-2684, Wincott et al., 1997, Methods Mol. Bio., 74, 59, Brennan et al., 1998, Biotechnol Bioeng., 61, 33-45, and Brennan, U.S. Pat. No. 6,001,311. All of these references are incorporated herein by reference. The synthesis of oligonucleotides makes use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5′-end, and phosphoramidites at the 3′-end. In a non-limiting example, small scale syntheses are conducted on a 394 Applied Biosystems, Inc. synthesizer using a 0.2 μmol scale protocol with a 2.5 min coupling step for 2′-O-methylated nucleotides and a 45 second coupling step for 2′-deoxy nucleotides or 2′-deoxy-2′-fluoro nucleotides. Table V outlines the amounts and the contact times of the reagents used in the synthesis cycle. Alternatively, syntheses at the 0.2 μmol scale can be performed on a 96-well plate synthesizer, such as the instrument produced by Protogene (Palo Alto, Calif.) with minimal modification to the cycle. A 33-fold excess (60 μL of 0.11 M=6.6 μmol) of 2′-O-methyl phosphoramidite and a 105-fold excess of S-ethyl tetrazole (60 μL of 0.25 M=15 μmol) can be used in each coupling cycle of 2′-O-methyl residues relative to polymer-bound 5′-hydroxyl. A 22-fold excess (40 μL of 0.11 M=4.4 μmol) of deoxy phosphoramidite and a 70-fold excess of S-ethyl tetrazole (40 μL of 0.25 M=10 μmol) can be used in each coupling cycle of deoxy residues relative to polymer-bound 5′-hydroxyl. Average coupling yields on the 394 Applied Biosystems, Inc. synthesizer, determined by colorimetric quantitation of the trityl fractions, are typically 97.5-99%. Other oligonucleotide synthesis reagents for the 394 Applied Biosystems, Inc. synthesizer include the following: detritylation solution is 3% TCA in methylene chloride (ABI); capping is performed with 16% N-methyl imidazole in THF (ABI) and 10% acetic anhydride/10% 2,6-lutidine in THF (ABI); and oxidation solution is 16.9 mM I₂, 49 mM pyridine, 9% water in THF (PerSeptive Biosystems, Inc.). Burdick & Jackson Synthesis Grade acetonitrile is used directly from the reagent bottle. S-Ethyltetrazole solution (0.25 M in acetonitrile) is made up from the solid obtained from American International Chemical, Inc. Alternately, for the introduction of phosphorothioate linkages, Beaucage reagent (3H-1,2-Benzodithiol-3-one 1,1-dioxide, 0.05 M in acetonitrile) is used.

Deprotection of the DNA-based oligonucleotides is performed as follows: the polymer-bound trityl-on oligoribonucleotide is transferred to a 4 mL glass screw top vial and suspended in a solution of 40% aqueous methylamine (1 mL) at 65° C. for 10 minutes. After cooling to −20° C., the supernatant is removed from the polymer support. The support is washed three times with 1.0 mL of EtOH:MeCN:H2O/3:1:1, vortexed and the supernatant is then added to the first supernatant. The combined supernatants, containing the oligoribonucleotide, are dried to a white powder.

The method of synthesis used for RNA including certain siNA molecules of the invention follows the procedure as described in Usman et al., 1987, J. Am. Chem. Soc., 109, 7845; Scaringe et al., 1990, Nucleic Acids Res., 18, 5433; and Wincott et al., 1995, Nucleic Acids Res. 23, 2677-2684 Wincott et al., 1997, Methods Mol. Bio., 74, 59, and makes use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5′-end, and phosphoramidites at the 3′-end. In a non-limiting example, small scale syntheses are conducted on a 394 Applied Biosystems, Inc. synthesizer using a 0.2 μmol scale protocol with a 7.5 min coupling step for alkylsilyl protected nucleotides and a 2.5 min coupling step for 2′-O-methylated nucleotides. Table V outlines the amounts and the contact times of the reagents used in the synthesis cycle. Alternatively, syntheses at the 0.2 μmol scale can be done on a 96-well plate synthesizer, such as the instrument produced by Protogene (Palo Alto, Calif.) with minimal modification to the cycle. A 33-fold excess (60 μL of 0.11 M=6.6 μmol) of 2′-O-methyl phosphoramidite and a 75-fold excess of S-ethyl tetrazole (60 μL of 0.25 M=15 μmol) can be used in each coupling cycle of 2′-O-methyl residues relative to polymer-bound 5′-hydroxyl. A 66-fold excess (120 μL of 0.11 M=13.2 μmol) of alkylsilyl (ribo) protected phosphoramidite and a 150-fold excess of S-ethyl tetrazole (120 μL of 0.25 M=30 μmol) can be used in each coupling cycle of ribo residues relative to polymer-bound 5′-hydroxyl. Average coupling yields on the 394 Applied Biosystems, Inc. synthesizer, determined by colorimetric quantitation of the trityl fractions, are typically 97.5-99%. Other oligonucleotide synthesis reagents for the 394 Applied Biosystems, Inc. synthesizer include the following: detritylation solution is 3% TCA in methylene chloride (ABI); capping is performed with 16% N-methyl imidazole in THF (ABI) and 10% acetic anhydride/10% 2,6-lutidine in THF (ABI); oxidation solution is 16.9 mM I₂, 49 mM pyridine, 9% water in THF (PerSeptive Biosystems, Inc.). Burdick & Jackson Synthesis Grade acetonitrile is used directly from the reagent bottle. S-Ethyltetrazole solution (0.25 M in acetonitrile) is made up from the solid obtained from American International Chemical, Inc. Alternately, for the introduction of phosphorothioate linkages, Beaucage reagent (3H-1,2-Benzodithiol-3-one 1,1-dioxide 0.05 M in acetonitrile) is used.

Deprotection of the RNA is performed using either a two-pot or one-pot protocol. For the two-pot protocol, the polymer-bound trityl-on oligoribonucleotide is transferred to a 4 mL glass screw top vial and suspended in a solution of 40% aq. methylamine (1 mL) at 65° C. for 10 min. After cooling to −20° C., the supernatant is removed from the polymer support. The support is washed three times with 1.0 mL of EtOH:MeCN:H2O/3:1:1, vortexed and the supernatant is then added to the first supernatant. The combined supernatants, containing the oligoribonucleotide, are dried to a white powder. The base deprotected oligoribonucleotide is resuspended in anhydrous TEA/HF/NMP solution (300 μL of a solution of 1.5 mL N-methylpyrrolidinone, 750 μL TEA and 1 mL TEA.3HF to provide a 1.4 M HF concentration) and heated to 65° C. After 1.5 h, the oligomer is quenched with 1.5 M NH₄HCO₃.

Alternatively, for the one-pot protocol, the polymer-bound trityl-on oligoribonucleotide is transferred to a 4 mL glass screw top vial and suspended in a solution of 33% ethanolic methylamine/DMSO: 1/1 (0.8 mL) at 65° C. for 15 minutes. The vial is brought to room temperature TEA.3HF (0.1 mL) is added and the vial is heated at 65° C. for 15 minutes. The sample is cooled at −20° C. and then quenched with 1.5 M NH₄HCO₃.

For purification of the trityl-on oligomers, the quenched NH₄HCO₃solution is loaded onto a C-18 containing cartridge that had been prewashed with acetonitrile followed by 50 mM TEAA. After washing the loaded cartridge with water, the RNA is detritylated with 0.5% TFA for 13 minutes. The cartridge is then washed again with water, salt exchanged with 1 M NaCl and washed with water again. The oligonucleotide is then eluted with 30% acetonitrile.

The average stepwise coupling yields are typically >98% (Wincott et al., 1995 Nucleic Acids Res. 23, 2677-2684). Those of ordinary skill in the art will recognize that the scale of synthesis can be adapted to be larger or smaller than the example described above including but not limited to 96-well format.

Alternatively, the nucleic acid molecules of the present invention can be synthesized separately and joined together post-synthetically, for example, by ligation (Moore et al., 1992, Science 256, 9923; Draper et al., International PCT publication No. WO 93/23569; Shabarova et al., 1991, Nucleic Acids Research 19, 4247; Bellon et al., 1997, Nucleosides & Nucleotides, 16, 951; Bellon et al., 1997, Bioconjugate Chem. 8, 204), or by hybridization following synthesis and/or deprotection.

The siNA molecules of the invention can also be synthesized via a tandem synthesis methodology as described in Example 1 herein, wherein both siNA strands are synthesized as a single contiguous oligonucleotide fragment or strand separated by a cleavable linker which is subsequently cleaved to provide separate siNA fragments or strands that hybridize and permit purification of the siNA duplex. The linker can be a polynucleotide linker or a non-nucleotide linker. The tandem synthesis of siNA as described herein can be readily adapted to both multiwell/multiplate synthesis platforms such as 96 well or similarly larger multi-well platforms. The tandem synthesis of siNA as described herein can also be readily adapted to large scale synthesis platforms employing batch reactors, synthesis columns and the like.

A siNA molecule can also be assembled from two distinct nucleic acid strands or fragments wherein one fragment includes the sense region and the second fragment includes the antisense region of the RNA molecule.

The nucleic acid molecules of the present invention can be modified extensively to enhance stability by modification with nuclease resistant groups, for example, 2′-amino, 2′-C-allyl, 2′-fluoro, 2′-O-methyl, 2′-H (for a review see Usman and Cedergren, 1992, TIBS 17, 34; Usman et al., 1994, Nucleic Acids Symp. Ser. 31, 163). siNA constructs can be purified by gel electrophoresis using general methods or can be purified by high pressure liquid chromatography (HPLC; see Wincott et al., supra, the totality of which is hereby incorporated herein by reference) and re-suspended in water.

In another aspect of the invention, siNA molecules of the invention are expressed from transcription units inserted into DNA or RNA vectors. The recombinant vectors can be DNA plasmids or viral vectors. siNA expressing viral vectors can be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. The recombinant vectors capable of expressing the siNA molecules can be delivered as described herein, and persist in target cells. Alternatively, viral vectors can be used that provide for transient expression of siNA molecules.

Optimizing Activity of the Nucleic Acid Molecule of the Invention.

Chemically synthesizing nucleic acid molecules with modifications (base, sugar and/or phosphate) can prevent their degradation by serum ribonucleases, which can increase their potency (see e.g., Eckstein et al., International Publication No. WO 92/07065; Perrault et al., 1990 Nature 344, 565; Pieken et al., 1991, Science 253, 314; Usman and Cedergren, 1992, Trends in Biochem. Sci. 17, 334; Usman et al., International Publication No. WO 93/15187; and Rossi et al., International Publication No. WO 91/03162; Sproat, U.S. Pat. No. 5,334,711; Gold et al., U.S. Pat. No. 6,300,074; and Burgin et al., supra; all of which are incorporated by reference herein). All of the above references describe various chemical modifications that can be made to the base, phosphate and/or sugar moieties of the nucleic acid molecules described herein. Modifications that enhance their efficacy in cells, and removal of bases from nucleic acid molecules to shorten oligonucleotide synthesis times and reduce chemical requirements are desired.

There are several examples in the art describing sugar, base and phosphate modifications that can be introduced into nucleic acid molecules with significant enhancement in their nuclease stability and efficacy. For example, oligonucleotides are modified to enhance stability and/or enhance biological activity by modification with nuclease resistant groups, for example, 2′-amino, 2′-C-allyl, 2′-fluoro, 2′-O-methyl, 2′-O-allyl, 2′-H, nucleotide base modifications (for a review see Usman and Cedergren, 1992, TIBS. 17, 34; Usman et al., 1994, Nucleic Acids Symp. Ser. 31, 163; Burgin et al., 1996, Biochemistry, 35, 14090). Sugar modification of nucleic acid molecules have been extensively described in the art (see Eckstein et al., International Publication PCT No. WO 92/07065; Perrault et al. Nature, 1990, 344, 565-568; Pieken et al. Science, 1991, 253, 314-317; Usman and Cedergren, Trends in Biochem. Sci., 1992, 17, 334-339; Usman et al. International Publication PCT No. WO 93/15187; Sproat, U.S. Pat. No. 5,334,711 and Beigelman et al., 1995, J. Biol. Chem., 270, 25702; Beigelman et al., International PCT publication No. WO 97/26270; Beigelman et al., U.S. Pat. No. 5,716,824; Usman et al., U.S. Pat. No. 5,627,053; Woolf et al., International PCT Publication No. WO 98/13526; Thompson et al., U.S. Ser. No. 60/082,404 which was filed on Apr. 20, 1998; Karpeisky et al., 1998, Tetrahedron Lett., 39, 1131; Earnshaw and Gait, 1998, Biopolymers (Nucleic Acid Sciences), 48, 39-55; Verma and Eckstein, 1998, Annu. Rev. Biochem., 67, 99-134; and Burlina et al., 1997, Bioorg. Med. Chem., 5, 1999-2010; all of the references are hereby incorporated in their totality by reference herein). Such publications describe general methods and strategies to determine the location of incorporation of sugar, base and/or phosphate modifications and the like into nucleic acid molecules without modulating catalysis, and are incorporated by reference herein. In view of such teachings, similar modifications can be used as described herein to modify the siNA nucleic acid molecules of the instant invention so long as the ability of siNA to promote RNAi is cells is not significantly inhibited.

While chemical modification of oligonucleotide internucleotide linkages with phosphorothioate, phosphorodithioate, and/or 5′-methylphosphonate linkages improves stability, excessive modifications can cause some toxicity or decreased activity. Therefore, when designing nucleic acid molecules, the amount of these internucleotide linkages should be minimized. The reduction in the concentration of these linkages should lower toxicity, resulting in increased efficacy and higher specificity of these molecules.

Short interfering nucleic acid (siNA) molecules having chemical modifications that maintain or enhance activity are provided. Such a nucleic acid is also generally more resistant to nucleases than an unmodified nucleic acid. Accordingly, the in vitro and/or in vivo activity should not be significantly lowered. In cases in which modulation is the goal, therapeutic nucleic acid molecules delivered exogenously should optimally be stable within cells until translation of the target RNA has been modulated long enough to reduce the levels of the undesirable protein. This period of time varies between hours to days depending upon the disease state. Improvements in the chemical synthesis of RNA and DNA (Wincott et al., 1995, Nucleic Acids Res. 23, 2677; Caruthers et al., 1992, Methods in Enzymology 211, 3-19 (incorporated by reference herein)) have expanded the ability to modify nucleic acid molecules by introducing nucleotide modifications to enhance their nuclease stability, as described above.

In one embodiment, nucleic acid molecules of the invention include one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) G-clamp nucleotides. A G-clamp nucleotide is a modified cytosine analog wherein the modifications confer the ability to hydrogen bond both Watson-Crick and Hoogsteen faces of a complementary guanine within a duplex, see for example Lin and Matteucci, 1998, J. Am. Chem. Soc., 120, 8531-8532. A single G-clamp analog substitution within an oligonucleotide can result in substantially enhanced helical thermal stability and mismatch discrimination when hybridized to complementary oligonucleotides. The inclusion of such nucleotides in nucleic acid molecules of the invention results in both enhanced affinity and specificity to nucleic acid targets, complementary sequences, or template strands. In another embodiment, nucleic acid molecules of the invention include one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) LNA “locked nucleic acid” nucleotides such as a 2′,4′-C methylene bicyclo nucleotide (see for example Wengel et al., International PCT Publication No. WO 00/66604 and WO 99/14226).

In another embodiment, the invention features conjugates and/or complexes of siNA molecules of the invention. Such conjugates and/or complexes can be used to facilitate delivery of siNA molecules into a biological system, such as a cell. The conjugates and complexes provided by the instant invention can impart therapeutic activity by transferring therapeutic compounds across cellular membranes, altering the pharmacokinetics, and/or modulating the localization of nucleic acid molecules of the invention. The present invention encompasses the design and synthesis of novel conjugates and complexes for the delivery of molecules, including, but not limited to, small molecules, lipids, cholesterol, phospholipids, nucleosides, nucleotides, nucleic acids, antibodies, toxins, negatively charged polymers and other polymers, for example proteins, peptides, hormones, carbohydrates, polyethylene glycols, or polyamines, across cellular membranes. In general, the transporters described are designed to be used either individually or as part of a multi-component system, with or without degradable linkers. These compounds are expected to improve delivery and/or localization of nucleic acid molecules of the invention into a number of cell types originating from different tissues, in the presence or absence of serum (see Sullenger and Cech, U.S. Pat. No. 5,854,038). Conjugates of the molecules described herein can be attached to biologically active molecules via linkers that are biodegradable, such as biodegradable nucleic acid linker molecules.

The term “biodegradable linker” as used herein, refers to a nucleic acid or non-nucleic acid linker molecule that is designed as a biodegradable linker to connect one molecule to another molecule, for example, a biologically active molecule to a siNA molecule of the invention or the sense and antisense strands of a siNA molecule of the invention. The biodegradable linker is designed such that its stability can be modulated for a particular purpose, such as delivery to a particular tissue or cell type. The stability of a nucleic acid-based biodegradable linker molecule can be modulated by using various chemistries, for example combinations of ribonucleotides, deoxyribonucleotides, and chemically-modified nucleotides, such as 2′-O-methyl, 2′-fluoro, 2′-amino, 2′-O-amino, 2′-C-allyl, 2′-O-allyl, and other 2′-modified or base modified nucleotides. The biodegradable nucleic acid linker molecule can be a dimer, trimer, tetramer or longer nucleic acid molecule, for example, an oligonucleotide of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides in length, or can comprise a single nucleotide with a phosphorus-based linkage, for example, a phosphoramidate or phosphodiester linkage. The biodegradable nucleic acid linker molecule can also comprise nucleic acid backbone, nucleic acid sugar, or nucleic acid base modifications.

The term “biodegradable” as used herein, refers to degradation in a biological system, for example, enzymatic degradation or chemical degradation.

The term “biologically active molecule” as used herein refers to compounds or molecules that are capable of eliciting or modifying a biological response in a system. Non-limiting examples of biologically active siNA molecules either alone or in combination with other molecules contemplated by the instant invention include therapeutically active molecules such as antibodies, cholesterol, hormones, antivirals, peptides, proteins, chemotherapeutics, small molecules, vitamins, co-factors, nucleosides, nucleotides, oligonucleotides, enzymatic nucleic acids, antisense nucleic acids, triplex forming oligonucleotides, 2,5-A chimeras, siNA, dsRNA, allozymes, aptamers, decoys and analogs thereof. Biologically active molecules of the invention also include molecules capable of modulating the pharmacokinetics and/or pharmacodynamics of other biologically active molecules, for example, lipids and polymers such as polyamines, polyamides, polyethylene glycol and other polyethers.

The term “phospholipid” as used herein, refers to a hydrophobic molecule comprising at least one phosphorus group. For example, a phospholipid can comprise a phosphorus-containing group and saturated or unsaturated alkyl group, optionally substituted with OH, COOH, oxo, amine, or substituted or unsubstituted aryl groups.

Therapeutic nucleic acid molecules (e.g., siNA molecules) delivered exogenously optimally are stable within cells until reverse transcription of the RNA has been modulated long enough to reduce the levels of the RNA transcript. The nucleic acid molecules are resistant to nucleases in order to function as effective intracellular therapeutic agents. Improvements in the chemical synthesis of nucleic acid molecules described in the instant invention and in the art have expanded the ability to modify nucleic acid molecules by introducing nucleotide modifications to enhance their nuclease stability as described above.

In yet another embodiment, siNA molecules having chemical modifications that maintain or enhance enzymatic activity of proteins involved in RNAi are provided. Such nucleic acids are also generally more resistant to nucleases than unmodified nucleic acids. Thus, in vitro and/or in vivo the activity should not be significantly lowered.

Use of the nucleic acid-based molecules of the invention will lead to better treatments by affording the possibility of combination therapies (e.g., multiple siNA molecules targeted to different genes; nucleic acid molecules coupled with known small molecule modulators; or intermittent treatment with combinations of molecules, including different motifs and/or other chemical or biological molecules). The treatment of subjects with siNA molecules can also include combinations of different types of nucleic acid molecules, such as enzymatic nucleic acid molecules (ribozymes), allozymes, antisense, 2,5-A oligoadenylate, decoys, and aptamers.

In another aspect a siNA molecule of the invention comprises one or more 5′ and/or a 3′-cap structure, for example, on only the sense siNA strand, the antisense siNA strand, or both siNA strands.

By “cap structure” is meant chemical modifications, which have been incorporated at either terminus of the oligonucleotide (see, for example, Adamic et al., U.S. Pat. No. 5,998,203, incorporated by reference herein). These terminal modifications protect the nucleic acid molecule from exonuclease degradation, and may help in delivery and/or localization within a cell. The cap may be present at the 5′-terminus (5′-cap) or at the 3′-terminal (3′-cap) or may be present on both termini. In non-limiting examples, the 5′-cap includes, but is not limited to, glyceryl, inverted deoxy abasic residue (moiety); 4′,5′-methylene nucleotide; 1-(beta-D-erythrofuranosyl) nucleotide, 4′-thio nucleotide; carbocyclic nucleotide; 1,5-anhydrohexitol nucleotide; L-nucleotides; alpha-nucleotides; modified base nucleotide; phosphorodithioate linkage; threo-pentofuranosyl nucleotide; acyclic 3′,4′-seco nucleotide; acyclic 3,4-dihydroxybutyl nucleotide; acyclic 3,5-dihydroxypentyl nucleotide, 3′-3′-inverted nucleotide moiety; 3′-3′-inverted abasic moiety; 3′-2′-inverted nucleotide moiety; 3′-2′-inverted abasic moiety; 1,4-butanediol phosphate; 3′-phosphoramidate; hexylphosphate; aminohexyl phosphate; 3′-phosphate; 3′-phosphorothioate; phosphorodithioate; or bridging or non-bridging methylphosphonate moiety. Non-limiting examples of cap moieties are shown in FIG. 10.

Non-limiting examples of the 3′-cap include, but are not limited to, glyceryl, inverted deoxy abasic residue (moiety), 4′,5′-methylene nucleotide; 1-(beta-D-erythrofuranosyl) nucleotide; 4′-thio nucleotide, carbocyclic nucleotide; 5′-amino-alkyl phosphate; 1,3-diamino-2-propyl phosphate; 3-aminopropyl phosphate; 6-aminohexyl phosphate; 1,2-aminododecyl phosphate; hydroxypropyl phosphate; 1,5-anhydrohexitol nucleotide; L-nucleotide; alpha-nucleotide; modified base nucleotide; phosphorodithioate; threo-pentofuranosyl nucleotide; acyclic 3′,4′-seco nucleotide; 3,4-dihydroxybutyl nucleotide; 3,5-dihydroxypentyl nucleotide, 5′-5′-inverted nucleotide moiety; 5′-5′-inverted abasic moiety; 5′-phosphoramidate; 5′-phosphorothioate; 1,4-butanediol phosphate; 5′-amino; bridging and/or non-bridging 5′-phosphoramidate, phosphorothioate and/or phosphorodithioate, bridging or non bridging methylphosphonate and 5′-mercapto moieties (for more details see Beaucage and Iyer, 1993, Tetrahedron 49, 1925; incorporated by reference herein).

By the term “non-nucleotide” is meant any group or compound which can be incorporated into a nucleic acid chain in the place of one or more nucleotide units, including either sugar and/or phosphate substitutions, and allows the remaining bases to exhibit their enzymatic activity. The group or compound is abasic in that it does not contain a commonly recognized nucleotide base, such as adenosine, guanine, cytosine, uracil or thymine and therefore lacks a base at the 1′-position.

An “alkyl” group refers to a saturated aliphatic hydrocarbon, including straight-chain, branched-chain, and cyclic alkyl groups. Preferably, the alkyl group has 1 to 12 carbons. More preferably, it is a lower alkyl of from 1 to 7 carbons, more preferably 1 to 4 carbons. The alkyl group can be substituted or unsubstituted. When substituted the substituted group(s) is preferably, hydroxyl, cyano, alkoxy, ═O, ═S, NO₂or N(CH₃)₂, amino, or SH. The term also includes alkenyl groups that are unsaturated hydrocarbon groups containing at least one carbon-carbon double bond, including straight-chain, branched-chain, and cyclic groups. Preferably, the alkenyl group has 1 to 12 carbons. More preferably, it is a lower alkenyl of from 1 to 7 carbons, more preferably 1 to 4 carbons. The alkenyl group may be substituted or unsubstituted. When substituted the substituted group(s) is preferably, hydroxyl, cyano, alkoxy, ═O, ═S, NO₂, halogen, N(CH₃)₂, amino, or SH. The term “alkyl” also includes alkynyl groups that have an unsaturated hydrocarbon group containing at least one carbon-carbon triple bond, including straight-chain, branched-chain, and cyclic groups. Preferably, the alkynyl group has 1 to 12 carbons. More preferably, it is a lower alkynyl of from 1 to 7 carbons, more preferably 1 to 4 carbons. The alkynyl group may be substituted or unsubstituted. When substituted the substituted group(s) is preferably, hydroxyl, cyano, alkoxy, ═O, ═S, NO₂or N(CH₃)₂, amino or SH.

Such alkyl groups can also include aryl, alkylaryl, carbocyclic aryl, heterocyclic aryl, amide and ester groups. An “aryl” group refers to an aromatic group that has at least one ring having a conjugated pi electron system and includes carbocyclic aryl, heterocyclic aryl and biaryl groups, all of which may be optionally substituted. The preferred substituent(s) of aryl groups are halogen, trihalomethyl, hydroxyl, SH, OH, cyano, alkoxy, alkyl, alkenyl, alkynyl, and amino groups. An “alkylaryl” group refers to an alkyl group (as described above) covalently joined to an aryl group (as described above). Carbocyclic aryl groups are groups wherein the ring atoms on the aromatic ring are all carbon atoms. The carbon atoms are optionally substituted. Heterocyclic aryl groups are groups having from 1 to 3 heteroatoms as ring atoms in the aromatic ring and the remainder of the ring atoms are carbon atoms. Suitable heteroatoms include oxygen, sulfur, and nitrogen, and include furanyl, thienyl, pyridyl, pyrrolyl, N-lower alkyl pyrrolo, pyrimidyl, pyrazinyl, imidazolyl and the like, all optionally substituted. An “amide” refers to an —C(O)—NH—R, where R is either alkyl, aryl, alkylaryl or hydrogen. An “ester” refers to an —C(O)—OR′, where R is either alkyl, aryl, alkylaryl or hydrogen.

By “nucleotide” as used herein is as recognized in the art to include natural bases (standard), and modified bases well known in the art. Such bases are generally located at the 1′ position of a nucleotide sugar moiety. Nucleotides generally comprise a base, sugar and a phosphate group. The nucleotides can be unmodified or modified at the sugar, phosphate and/or base moiety, (also referred to interchangeably as nucleotide analogs, modified nucleotides, non-natural nucleotides, non-standard nucleotides and other; see, for example, Usman and McSwiggen, supra; Eckstein et al., International PCT Publication No. WO 92/07065; Usman et al., International PCT Publication No. WO 93/15187; Uhlman & Peyman, supra, all are hereby incorporated by reference herein). There are several examples of modified nucleic acid bases known in the art as summarized by Limbach et al., 1994, Nucleic Acids Res. 22, 2183. Some of the non-limiting examples of base modifications that can be introduced into nucleic acid molecules include, inosine, purine, pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2,4,6-trimethoxy benzene, 3-methyl uracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidines (e.g., 5-methylcytidine), 5-alkyluridines (e.g., ribothymidine), 5-halouridine (e.g., 5-bromouridine) or 6-azapyrimidines or 6-alkylpyrimidines (e.g. 6-methyluridine), propyne, and others (Burgin et al., 1996, Biochemistry, 35, 14090; Uhlman & Peyman, supra). By “modified bases” in this aspect is meant nucleotide bases other than adenine, guanine, cytosine and uracil at 1′ position or their equivalents.

In one embodiment, the invention features modified siNA molecules, with phosphate backbone modifications comprising one or more phosphorothioate, phosphorodithioate, methylphosphonate, phosphotriester, morpholino, amidate carbamate, carboxymethyl, acetamidate, polyamide, sulfonate, sulfonamide, sulfamate, formacetal, thioformacetal, and/or alkylsilyl, substitutions. For a review of oligonucleotide backbone modifications, see Hunziker and Leumann, 1995, Nucleic Acid Analogues: Synthesis and Properties, in Modern Synthetic Methods, VCH, 331-417, and Mesmaeker et al., 1994, Novel Backbone Replacements for Oligonucleotides, in Carbohydrate Modifications in Antisense Research, ACS, 24-39.

By “abasic” is meant sugar moieties lacking a nucleobase or having a hydrogen atom (H) or other other non-nucleobase chemical groups in place of a nucleobase at the 1′ position of the sugar moiety, see for example Adamic et al., U.S. Pat. No. 5,998,203. In one embodiment, an abasic moiety of the invention is a ribose, deoxyribose, or dideoxyribose sugar.

By “unmodified nucleoside” is meant one of the bases adenine, cytosine, guanine, thymine, or uracil joined to the 1′ carbon of β-D-ribo-furanose.

By “modified nucleoside” is meant any nucleotide base which contains a modification in the chemical structure of an unmodified nucleotide base, sugar and/or phosphate. Non-limiting examples of modified nucleotides are shown by Formulae I-VII and/or other modifications described herein.

In connection with 2′-modified nucleotides as described for the present invention, by “amino” is meant 2′-NH₂or 2′-O—NH₂, which can be modified or unmodified. Such modified groups are described, for example, in Eckstein et al., U.S. Pat. No. 5,672,695 and Matulic-Adamic et al., U.S. Pat. No. 6,248,878, which are both incorporated by reference in their entireties.

Various modifications to nucleic acid siNA structure can be made to enhance the utility of these molecules. Such modifications will enhance shelf-life, half-life in vitro, stability, and ease of introduction of such oligonucleotides to the target site, e.g., to enhance penetration of cellular membranes, and confer the ability to recognize and bind to targeted cells.

Administration of Nucleic Acid Molecules

A siNA molecule of the invention can be adapted for use to prevent or treat diseases, traits, disorders, and/or conditions described herein or otherwise known in the art to be related to gene expression, and/or any other trait, disease, disorder or condition that is related to or will respond to the levels of a target polynucleotide or a protein expressed therefrom in a cell or tissue, alone or in combination with other therapies.

In one embodiment, a siNA composition of the invention can comprise a delivery vehicle, including liposomes, for administration to a subject, carriers and diluents and their salts, and/or can be present in pharmaceutically acceptable formulations. Methods for the delivery of nucleic acid molecules are described in Akhtar et al., 1992, Trends Cell Bio., 2, 139; Delivery Strategies for Antisense Oligonucleotide Therapeutics, ed. Akhtar, 1995, Maurer et al., 1999, Mol. Membr. Biol., 16, 129-140; Hofland and Huang, 1999, Handb. Exp. Pharmacol., 137, 165-192; and Lee et al., 2000, ACS Symp. Ser., 752, 184-192, all of which are incorporated herein by reference. Beigelman et al., U.S. Pat. No. 6,395,713 and Sullivan et al., PCT WO 94/02595 further describe the general methods for delivery of nucleic acid molecules. These protocols can be utilized for the delivery of virtually any nucleic acid molecule. Nucleic acid molecules can be administered to cells by a variety of methods known to those of skill in the art, including, but not restricted to, encapsulation in liposomes, by iontophoresis, or by incorporation into other vehicles, such as biodegradable polymers, hydrogels, cyclodextrins (see for example Gonzalez et al., 1999, Bioconjugate Chem., 10, 1068-1074; Wang et al., International PCT publication Nos. WO 03/47518 and WO 03/46185), poly(lactic-co-glycolic)acid (PLGA) and PLCA microspheres (see for example U.S. Pat. No. 6,447,796 and US Patent Application Publication No. US 2002130430), biodegradable nanocapsules, and bioadhesive microspheres, or by proteinaceous vectors (O'Hare and Normand, International PCT Publication No. WO 00/53722). In another embodiment, the nucleic acid molecules of the invention can also be formulated or complexed with polyethyleneimine and derivatives thereof, such as polyethyleneimine-polyethyleneglycol-N-acetylgalactosamine (PEI-PEG-GAL) or polyethyleneimine-polyethyleneglycol-tri-N-acetylgalactosamine (PEI-PEG-triGAL) derivatives. In one embodiment, the nucleic acid molecules of the invention are formulated as described in United States Patent Application Publication No. 20030077829, incorporated by reference herein in its entirety.

In one embodiment, a siNA molecule of the invention is formulated as a composition described in U.S. Provisional patent application No. 60/678,531 and in related U.S. Provisional patent application No. TBD, filed Jul. 29, 2005 (Vargeese et al.), both of which are incorporated by reference herein in their entirety. Such siNA formuations are generally referred to as “lipid nucleic acid particles” (LNP).

In one embodiment, a siNA molecule of the invention is complexed with membrane disruptive agents such as those described in U.S. Patent Application Publication No. 20010007666, incorporated by reference herein in its entirety including the drawings. In another embodiment, the membrane disruptive agent or agents and the siNA molecule are also complexed with a cationic lipid or helper lipid molecule, such as those lipids described in U.S. Pat. No. 6,235,310, incorporated by reference herein in its entirety including the drawings.

In one embodiment, a siNA molecule of the invention is complexed with delivery systems as described in U.S. Patent Application Publication No. 2003077829 and International PCT Publication Nos. WO 00/03683 and WO 02/087541, all incorporated by reference herein in their entirety including the drawings.

In one embodiment, the nucleic acid molecules of the invention are administered to skeletal tissues (e.g., bone, cartilage, tendon, ligament) or bone metastatic tumors via atelocollagen complexation or conjugation (see for example Takeshita et al., 2005, PNAS, 102, 12177-12182). Therefore, in one embodiment, the instant invention features one or more dsiNA molecules as a composition complexed with atelocollagen. In another embodiment, the instant invention features one or more siNA molecules conjugated to atelocollagen via a linker as described herein or otherwise known in the art.

In one embodiment, the nucleic acid molecules of the invention are administered via pulmonary delivery, such as by inhalation of an aerosol or spray dried formulation administered by an inhalation device or nebulizer, providing rapid local uptake of the nucleic acid molecules into relevant pulmonary tissues. Solid particulate compositions containing respirable dry particles of micronized nucleic acid compositions can be prepared by grinding dried or lyophilized nucleic acid compositions, and then passing the micronized composition through, for example, a 400 mesh screen to break up or separate out large agglomerates. A solid particulate composition comprising the nucleic acid compositions of the invention can optionally contain a dispersant which serves to facilitate the formation of an aerosol as well as other therapeutic compounds. A suitable dispersant is lactose, which can be blended with the nucleic acid compound in any suitable ratio, such as a 1 to 1 ratio by weight.

Aerosols of liquid particles comprising a nucleic acid composition of the invention can be produced by any suitable means, such as with a nebulizer (see for example U.S. Pat. No. 4,501,729). Nebulizers are commercially available devices which transform solutions or suspensions of an active ingredient into a therapeutic aerosol mist either by means of acceleration of a compressed gas, typically air or oxygen, through a narrow venturi orifice or by means of ultrasonic agitation. Suitable formulations for use in nebulizers comprise the active ingredient in a liquid carrier in an amount of up to 40% w/w preferably less than 20% w/w of the formulation. The carrier is typically water or a dilute aqueous alcoholic solution, preferably made isotonic with body fluids by the addition of, for example, sodium chloride or other suitable salts. Optional additives include preservatives if the formulation is not prepared sterile, for example, methyl hydroxybenzoate, anti-oxidants, flavorings, volatile oils, buffering agents and emulsifiers and other formulation surfactants. The aerosols of solid particles comprising the active composition and surfactant can likewise be produced with any solid particulate aerosol generator. Aerosol generators for administering solid particulate therapeutics to a subject produce particles which are respirable, as explained above, and generate a volume of aerosol containing a predetermined metered dose of a therapeutic composition at a rate suitable for human administration.

In one embodiment, a solid particulate aerosol generator of the invention is an insufflator. Suitable formulations for administration by insufflation include finely comminuted powders which can be delivered by means of an insufflator. In the insufflator, the powder, e.g., a metered dose thereof effective to carry out the treatments described herein, is contained in capsules or cartridges, typically made of gelatin or plastic, which are either pierced or opened in situ and the powder delivered by air drawn through the device upon inhalation or by means of a manually-operated pump. The powder employed in the insufflator consists either solely of the active ingredient or of a powder blend comprising the active ingredient, a suitable powder diluent, such as lactose, and an optional surfactant. The active ingredient typically comprises from 0.1 to 100 w/w of the formulation. A second type of illustrative aerosol generator comprises a metered dose inhaler. Metered dose inhalers are pressurized aerosol dispensers, typically containing a suspension or solution formulation of the active ingredient in a liquified propellant. During use these devices discharge the formulation through a valve adapted to deliver a metered volume to produce a fine particle spray containing the active ingredient. Suitable propellants include certain chlorofluorocarbon compounds, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane and mixtures thereof. The formulation can additionally contain one or more co-solvents, for example, ethanol, emulsifiers and other formulation surfactants, such as oleic acid or sorbitan trioleate, anti-oxidants and suitable flavoring agents. Other methods for pulmonary delivery are described in, for example US Patent Application No. 20040037780, and U.S. Pat. Nos. 6,592,904; 6,582,728; 6,565,885, all incorporated by reference herein.

In one embodiment, the invention features the use of methods to deliver the nucleic acid molecules of the instant invention to the central nervous system and/or peripheral nervous system. Experiments have demonstrated the efficient in vivo uptake of nucleic acids by neurons. As an example of local administration of nucleic acids to nerve cells, Sommer et al., 1998, Antisense Nuc. Acid Drug Dev., 8, 75, describe a study in which a 15mer phosphorothioate antisense nucleic acid molecule to c-fos is administered to rats via microinjection into the brain. Antisense molecules labeled with tetramethylrhodamine-isothiocyanate (TRITC) or fluorescein isothiocyanate (FITC) were taken up by exclusively by neurons thirty minutes post-injection. A diffuse cytoplasmic staining and nuclear staining was observed in these cells. As an example of systemic administration of nucleic acid to nerve cells, Epa et al., 2000, Antisense Nuc. Acid Drug Dev., 10, 469, describe an in vivo mouse study in which beta-cyclodextrin-adamantane-oligonucleotide conjugates were used to target the p75 neurotrophin receptor in neuronally differentiated PC12 cells. Following a two week course of IP administration, pronounced uptake of p75 neurotrophin receptor antisense was observed in dorsal root ganglion (DRG) cells. In addition, a marked and consistent down-regulation of p75 was observed in DRG neurons. Additional approaches to the targeting of nucleic acid to neurons are described in Broaddus et al., 1998, J. Neurosurg., 88(4), 734; Karle et al., 1997, Eur. J. Pharmocol., 340(2/3), 153; Bannai et al., 1998, Brain Research, 784(1,2), 304; Rajakumar et al., 1997, Synapse, 26(3), 199; Wu-pong et al., 1999, BioPharm, 12(1), 32; Bannai et al., 1998, Brain Res. Protoc., 3(1), 83; Simantov et al., 1996, Neuroscience, 74(1), 39. Nucleic acid molecules of the invention are therefore amenable to delivery to and uptake by cells that express repeat expansion allelic variants for modulation of RE gene expression. The delivery of nucleic acid molecules of the invention, targeting RE is provided by a variety of different strategies. Traditional approaches to CNS delivery that can be used include, but are not limited to, intrathecal and intracerebroventricular administration, implantation of catheters and pumps, direct injection or perfusion at the site of injury or lesion, injection into the brain arterial system, or by chemical or osmotic opening of the blood-brain barrier. Other approaches can include the use of various transport and carrier systems, for example though the use of conjugates and biodegradable polymers. Furthermore, gene therapy approaches, for example as described in Kaplitt et al., U.S. Pat. No. 6,180,613 and Davidson, WO 04/013280, can be used to express nucleic acid molecules in the CNS.

The delivery of nucleic acid molecules of the invention to the CNS is provided by a variety of different strategies. Traditional approaches to CNS delivery that can be used include, but are not limited to, intrathecal and intracerebroventricular administration, implantation of catheters and pumps, direct injection or perfusion at the site of injury or lesion, injection into the brain arterial system, or by chemical or osmotic opening of the blood-brain barrier. Other approaches can include the use of various transport and carrier systems, for example though the use of conjugates and biodegradable polymers. Furthermore, gene therapy approaches, for example as described in Kaplitt et al., U.S. Pat. No. 6,180,613 and Davidson, WO 04/013280, can be used to express nucleic acid molecules in the CNS.

In one embodiment, a compound, molecule, or composition for the treatment of ocular conditions (e.g., macular degeneration, diabetic retinopathy etc.) is administered to a subject intraocularly or by intraocular means. In another embodiment, a compound, molecule, or composition for the treatment of ocular conditions (e.g., macular degeneration, diabetic retinopathy etc.) is administered to a subject periocularly or by periocular means (see for example Ahlheim et al., International PCT publication No. WO 03/24420). In one embodiment, a siNA molecule and/or formulation or composition thereof is administered to a subject intraocularly or by intraocular means. In another embodiment, a siNA molecule and/or formualtion or composition thereof is administered to a subject periocularly or by periocular means. Periocular administration generally provides a less invasive approach to administering siNA molecules and formualtion or composition thereof to a subject (see for example Ahlheim et al., International PCT publication No. WO 03/24420). The use of periocular administraction also minimizes the risk of retinal detachment, allows for more frequent dosing or administraction, provides a clinically relevant route of administraction for macular degeneration and other optic conditions, and also provides the possiblilty of using resevoirs (e.g., implants, pumps or other devices) for drug delivery. In one embodiment, siNA compounds and compositions of the invention are administered locally, e.g., via intraocular or periocular means, such as injection, iontophoresis (see, for example, WO 03/043689 and WO 03/030989), or implant, about every 1-50 weeks (e.g., about every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 weeks), alone or in combination with other comounds and/or therapeis herein. In one embodiment, siNA compounds and compositions of the invention are administered systemically (e.g., via intravenous, subcutaneous, intramuscular, infusion, pump, implant etc.) about every 1-50 weeks (e.g., about every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 weeks), alone or in combination with other comounds and/or therapies described herein and/or otherwise known in the art.

In one embodiment, a siNA molecule of the invention is administered iontophoretically, for example to a particular organ or compartment (e.g., the eye, back of the eye, heart, liver, kidney, bladder, prostate, tumor, CNS etc.). Non-limiting examples of iontophoretic delivery are described in, for example, WO 03/043689 and WO 03/030989, which are incorporated by reference in their entireties herein.

In one embodiment, the siNA molecules of the invention and formulations or compositions thereof are administered to the liver as is generally known in the art (see for example Wen et al., 2004, World J Gastroenterol., 10, 244-9; Murao et al., 2002, Pharm Res., 19, 1808-14; Liu et al., 2003, Gene Ther., 10, 180-7; Hong et al., 2003, J Pharm Pharmacol., 54, 51-8; Herrmann et al., 2004, Arch Virol., 149, 1611-7; and Matsuno et al., 2003, Gene Ther., 10, 1559-66).

In one embodiment, the invention features the use of methods to deliver the nucleic acid molecules of the instant invention to hematopoietic cells, including monocytes and lymphocytes. These methods are described in detail by Hartmann et al., 1998, J. Phamacol. Exp. Ther., 285(2), 920-928; Kronenwett et al., 1998, Blood, 91(3), 852-862; Filion and Phillips, 1997, Biochim. Biophys. Acta., 1329(2), 345-356; Ma and Wei, 1996, Leuk. Res., 20(11/12), 925-930; and Bongartz et al., 1994, Nucleic Acids Research, 22(22), 4681-8. Such methods, as described above, include the use of free oligonucleitide, cationic lipid formulations, liposome formulations including pH sensitive liposomes and immunoliposomes, and bioconjugates including oligonucleotides conjugated to fusogenic peptides, for the transfection of hematopoietic cells with oligonucleotides.

In one embodiment, the siNA molecules and compositions of the invention are administered to the inner ear by contacting the siNA with inner ear cells, tissues, or structures such as the cochlea, under conditions suitable for the administration. In one embodiment, the administration comprises methods and devices as described in U.S. Pat. Nos. 5,421,818, 5,476,446, 5,474,529, 6,045,528, 6,440,102, 6,685,697, 6,120,484; and 5,572,594; all incorporated by reference herein and the teachings of Silverstein, 1999, Ear Nose Throat J., 78, 595-8, 600; and Jackson and Silverstein, 2002, Otolaryngol Clin North Am., 35, 639-53, and adapted for use the siNA molecules of the invention.

In one embodiment, the siNA molecules of the invention and formulations or compositions thereof are administered directly or topically (e.g., locally) to the dermis or follicles as is generally known in the art (see for example Brand, 2001, Curr. Opin. Mol. Ther., 3, 244-8; Regnier et al., 1998, J. Drug Target, 5, 275-89; Kanikkannan, 2002, BioDrugs, 16, 339-47; Wraight et al., 2001, Pharmacol. Ther., 90, 89-104; and Preat and Dujardin, 2001, STP PharmaSciences, 11, 57-68). In one embodiment, the siNA molecules of the invention and formulations or compositions thereof are administered directly or topically using a hydroalcoholic gel formulation comprising an alcohol (e.g., ethanol or isopropanol), water, and optionally including additional agents such isopropyl myristate and carbomer 980.

In one embodiment, delivery systems of the invention include, for example, aqueous and nonaqueous gels, creams, multiple emulsions, microemulsions, liposomes, ointments, aqueous and nonaqueous solutions, lotions, aerosols, hydrocarbon bases and powders, and can contain excipients such as solubilizers, permeation enhancers (e.g., fatty acids, fatty acid esters, fatty alcohols and amino acids), and hydrophilic polymers (e.g., polycarbophil and polyvinylpyrolidone). In one embodiment, the pharmaceutically acceptable carrier is a liposome or a transdermal enhancer. Examples of liposomes which can be used in this invention include the following: (1) CellFectin, 1:1.5 (M/M) liposome formulation of the cationic lipid N,NI,NII,NIII-tetramethyl-N,NI,NII,NIII-tetrapalmit-y-spermine and dioleoyl phosphatidylethanolamine (DOPE) (GIBCO BRL); (2) Cytofectin GSV, 2:1 (M/M) liposome formulation of a cationic lipid and DOPE (Glen Research); (3) DOTAP (N-[1-(2,3-dioleoyloxy)-N,N,N-tri-methyl-ammoniummethylsulfate) (Boehringer Manheim); and (4) Lipofectamine, 3:1 (M/M) liposome formulation of the polycationic lipid DOSPA and the neutral lipid DOPE (GIBCO BRL).

In one embodiment, delivery systems of the invention include patches, tablets, suppositories, pessaries, gels and creams, and can contain excipients such as solubilizers and enhancers (e.g., propylene glycol, bile salts and amino acids), and other vehicles (e.g., polyethylene glycol, fatty acid esters and derivatives, and hydrophilic polymers such as hydroxypropylmethylcellulose and hyaluronic acid).

In one embodiment, a siNA molecule of the invention is administered iontophoretically, for example to the dermis or to other relevant tissues such as the inner ear/cochlea. Non-limiting examples of iontophoretic delivery are described in, for example, WO 03/043689 and WO 03/030989, which are incorporated by reference in their entireties herein.

In one embodiment, siNA molecules of the invention are formulated or complexed with polyethylenimine (e.g., linear or branched PEI) and/or polyethylenimine derivatives, including for example grafted PEIs such as galactose PEI, cholesterol PEI, antibody derivatized PEI, and polyethylene glycol PEI (PEG-PEI) derivatives thereof (see for example Ogris et al., 2001, AAPA PharmSci, 3, 1-11; Furgeson et al., 2003, Bioconjugate Chem., 14, 840-847; Kunath et al., 2002, Phramaceutical Research, 19, 810-817; Choi et al., 2001, Bull. Korean Chem. Soc., 22, 46-52; Bettinger et al., 1999, Bioconjugate Chem., 10, 558-561; Peterson et al., 2002, Bioconjugate Chem., 13, 845-854; Erbacher et al., 1999, Journal of Gene Medicine Preprint, 1, 1-18; Godbey et al., 1999., PNAS USA, 96, 5177-5181; Godbey et al., 1999, Journal of Controlled Release, 60, 149-160; Diebold et al., 1999, Journal of Biological Chemistry, 274, 19087-19094; Thomas and Klibanov, 2002, PNAS USA, 99, 14640-14645; and Sagara, U.S. Pat. No. 6,586,524, incorporated by reference herein.

In one embodiment, a siNA molecule of the invention comprises a bioconjugate, for example a nucleic acid conjugate as described in Vargeese et al., U.S. Ser. No. 10/427,160, filed Apr. 30, 2003; U.S. Pat. No. 6,528,631; U.S. Pat. No. 6,335,434; U.S. Pat. No. 6,235,886; U.S. Pat. No. 6,153,737; U.S. Pat. No. 5,214,136; U.S. Pat. No. 5,138,045, all incorporated by reference herein.

Thus, the invention features a pharmaceutical composition comprising one or more nucleic acid(s) of the invention in an acceptable carrier, such as a stabilizer, buffer, and the like. The polynucleotides of the invention can be administered (e.g., RNA, DNA or protein) and introduced to a subject by any standard means, with or without stabilizers, buffers, and the like, to form a pharmaceutical composition. When it is desired to use a liposome delivery mechanism, standard protocols for formation of liposomes can be followed. The compositions of the present invention can also be formulated and used as creams, gels, sprays, oils and other suitable compositions for topical, dermal, or transdermal administration as is known in the art.

The present invention also includes pharmaceutically acceptable formulations of the compounds described. These formulations include salts of the above compounds, e.g., acid addition salts, for example, salts of hydrochloric, hydrobromic, acetic acid, and benzene sulfonic acid.

A pharmacological composition or formulation refers to a composition or formulation in a form suitable for administration, e.g., systemic or local administration, into a cell or subject, including for example a human. Suitable forms, in part, depend upon the use or the route of entry, for example oral, transdermal, or by injection. Such forms should not prevent the composition or formulation from reaching a target cell (i.e., a cell to which the negatively charged nucleic acid is desirable for delivery). For example, pharmacological compositions injected into the blood stream should be soluble. Other factors are known in the art, and include considerations such as toxicity and forms that prevent the composition or formulation from exerting its effect.

In one embodiment, siNA molecules of the invention are administered to a subject by systemic administration in a pharmaceutically acceptable composition or formulation. By “systemic administration” is meant in vivo systemic absorption or accumulation of drugs in the blood stream followed by distribution throughout the entire body. Administration routes that lead to systemic absorption include, without limitation: intravenous, subcutaneous, portal vein, intraperitoneal, inhalation, oral, intrapulmonary and intramuscular. Each of these administration routes exposes the siNA molecules of the invention to an accessible diseased tissue. The rate of entry of a drug into the circulation has been shown to be a function of molecular weight or size. The use of a liposome or other drug carrier comprising the compounds of the instant invention can potentially localize the drug, for example, in certain tissue types, such as the tissues of the reticular endothelial system (RES). A liposome formulation that can facilitate the association of drug with the surface of cells, such as, lymphocytes and macrophages is also useful. This approach can provide enhanced delivery of the drug to target cells by taking advantage of the specificity of macrophage and lymphocyte immune recognition of abnormal cells.

By “pharmaceutically acceptable formulation” or “pharmaceutically acceptable composition” is meant, a composition or formulation that allows for the effective distribution of the nucleic acid molecules of the instant invention in the physical location most suitable for their desired activity. Non-limiting examples of agents suitable for formulation with the nucleic acid molecules of the instant invention include: P-glycoprotein inhibitors (such as Pluronic P85),; biodegradable polymers, such as poly (DL-lactide-coglycolide) microspheres for sustained release delivery (Emerich, D F et al, 1999, Cell Transplant, 8, 47-58); and loaded nanoparticles, such as those made of polybutylcyanoacrylate. Other non-limiting examples of delivery strategies for the nucleic acid molecules of the instant invention include material described in Boado et al., 1998, J. Pharm. Sci., 87, 1308-1315; Tyler et al., 1999, FEBS Lett., 421, 280-284; Pardridge et al., 1995, PNAS USA., 92, 5592-5596; Boado, 1995, Adv. Drug Delivery Rev., 15, 73-107; Aldrian-Herrada et al., 1998, Nucleic Acids Res., 26, 4910-4916; and Tyler et al., 1999, PNAS USA., 96, 7053-7058.

The invention also features the use of a composition comprising surface-modified liposomes containing poly (ethylene glycol) lipids (PEG-modified, or long-circulating liposomes or stealth liposomes) and nucleic acid molecules of the invention. These formulations offer a method for increasing the accumulation of drugs (e.g., siNA) in target tissues. This class of drug carriers resists opsonization and elimination by the mononuclear phagocytic system (MPS or RES), thereby enabling longer blood circulation times and enhanced tissue exposure for the encapsulated drug (Lasic et al. Chem. Rev. 1995, 95, 2601-2627; Ishiwata et al., Chem. Pharm. Bull. 1995, 43, 1005-1011). Such liposomes have been shown to accumulate selectively in tumors, presumably by extravasation and capture in the neovascularized target tissues (Lasic et al., Science 1995, 267, 1275-1276; Oku et al., 1995, Biochim. Biophys. Acta, 1238, 86-90). The long-circulating liposomes enhance the pharmacokinetics and pharmacodynamics of DNA and RNA, particularly compared to conventional cationic liposomes which are known to accumulate in tissues of the MPS (Liu et al., J. Biol. Chem. 1995, 42, 24864-24870; Choi et al., International PCT Publication No. WO 96/10391; Ansell et al., International PCT Publication No. WO 96/10390; Holland et al., International PCT Publication No. WO 96/10392). Long-circulating liposomes are also likely to protect drugs from nuclease degradation to a greater extent compared to cationic liposomes, based on their ability to avoid accumulation in metabolically aggressive MPS tissues such as the liver and spleen.

The present invention also includes compositions prepared for storage or administration that include a pharmaceutically effective amount of the desired compounds in a pharmaceutically acceptable carrier or diluent. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A. R. Gennaro edit. 1985), hereby incorporated by reference herein. For example, preservatives, stabilizers, dyes and flavoring agents can be provided. These include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid. In addition, antioxidants and suspending agents can be used.

A pharmaceutically effective dose is that dose required to prevent, inhibit the occurrence, or treat (alleviate a symptom to some extent, preferably all of the symptoms) of a disease state. The pharmaceutically effective dose depends on the type of disease, the composition used, the route of administration, the type of mammal being treated, the physical characteristics of the specific mammal under consideration, concurrent medication, and other factors that those skilled in the medical arts will recognize. Generally, an amount between 0.1 mg/kg and 100 mg/kg body weight/day of active ingredients is administered dependent upon potency of the negatively charged polymer.

The nucleic acid molecules of the invention and formulations thereof can be administered orally, topically, parenterally, by inhalation or spray, or rectally in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and/or vehicles. The term parenteral as used herein includes percutaneous, subcutaneous, intravascular (e.g., intravenous), intramuscular, or intrathecal injection or infusion techniques and the like. In addition, there is provided a pharmaceutical formulation comprising a nucleic acid molecule of the invention and a pharmaceutically acceptable carrier. One or more nucleic acid molecules of the invention can be present in association with one or more non-toxic pharmaceutically acceptable carriers and/or diluents and/or adjuvants, and if desired other active ingredients. The pharmaceutical compositions containing nucleic acid molecules of the invention can be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion, hard or soft capsules, or syrups or elixirs.

Compositions intended for oral use can be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions can contain one or more such sweetening agents, flavoring agents, coloring agents or preservative agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for the manufacture of tablets. These excipients can be, for example, inert diluents; such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example starch, gelatin or acacia; and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets can be uncoated or they can be coated by known techniques. In some cases such coatings can be prepared by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monosterate or glyceryl distearate can be employed.

Formulations for oral use can also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin or olive oil.

Aqueous suspensions contain the active materials in a mixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydropropyl-methylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents can be a naturally-occurring phosphatide, for example, lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions can also contain one or more preservatives, for example ethyl, or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

Oily suspensions can be formulated by suspending the active ingredients in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions can contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents and flavoring agents can be added to provide palatable oral preparations. These compositions can be preserved by the addition of an anti-oxidant such as ascorbic acid

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents or suspending agents are exemplified by those already mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, can also be present.

Pharmaceutical compositions of the invention can also be in the form of oil-in-water emulsions. The oily phase can be a vegetable oil or a mineral oil or mixtures of these. Suitable emulsifying agents can be naturally-occurring gums, for example gum acacia or gum tragacanth, naturally-occurring phosphatides, for example soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol, anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions can also contain sweetening and flavoring agents.

Syrups and elixirs can be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol, glucose or sucrose. Such formulations can also contain a demulcent, a preservative and flavoring and coloring agents. The pharmaceutical compositions can be in the form of a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents that have been mentioned above. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parentally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The nucleic acid molecules of the invention can also be administered in the form of suppositories, e.g., for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter and polyethylene glycols.

Nucleic acid molecules of the invention can be administered parenterally in a sterile medium. The drug, depending on the vehicle and concentration used, can either be suspended or dissolved in the vehicle. Advantageously, adjuvants such as local anesthetics, preservatives and buffering agents can be dissolved in the vehicle.

Dosage levels of the order of from about 0.1 mg to about 140 mg per kilogram of body weight per day are useful in the treatment of the above-indicated conditions (about 0.5 mg to about 7 g per subject per day). The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form varies depending upon the host treated and the particular mode of administration. Dosage unit forms generally contain between from about 1 mg to about 500 mg of an active ingredient.

It is understood that the specific dose level for any particular subject depends upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, and rate of excretion, drug combination and the severity of the particular disease undergoing therapy.

For administration to non-human animals, the composition can also be added to the animal feed or drinking water. It can be convenient to formulate the animal feed and drinking water compositions so that the animal takes in a therapeutically appropriate quantity of the composition along with its diet. It can also be convenient to present the composition as a premix for addition to the feed or drinking water.

The nucleic acid molecules of the present invention can also be administered to a subject in combination with other therapeutic compounds to increase the overall therapeutic effect. The use of multiple compounds to treat an indication can increase the beneficial effects while reducing the presence of side effects.

In one embodiment, the invention comprises compositions suitable for administering nucleic acid molecules of the invention to specific cell types. For example, the asialoglycoprotein receptor (ASGPr) (Wu and Wu, 1987, J. Biol. Chem. 262, 4429-4432) is unique to hepatocytes and binds branched galactose-terminal glycoproteins, such as asialoorosomucoid (ASOR). In another example, the folate receptor is overexpressed in many cancer cells. Binding of such glycoproteins, synthetic glycoconjugates, or folates to the receptor takes place with an affinity that strongly depends on the degree of branching of the oligosaccharide chain, for example, triatennary structures are bound with greater affinity than biatenarry or monoatennary chains (Baenziger and Fiete, 1980, Cell, 22, 611-620; Connolly et al., 1982, J. Biol. Chem., 257, 939-945). Lee and Lee, 1987, Glycoconjugate J., 4, 317-328, obtained this high specificity through the use of N-acetyl-D-galactosamine as the carbohydrate moiety, which has higher affinity for the receptor, compared to galactose. This “clustering effect” has also been described for the binding and uptake of mannosyl-terminating glycoproteins or glycoconjugates (Ponpipom et al., 1981, J. Med. Chem., 24, 1388-1395). The use of galactose, galactosamine, or folate based conjugates to transport exogenous compounds across cell membranes can provide a targeted delivery approach to, for example, the treatment of liver disease, cancers of the liver, or other cancers. The use of bioconjugates can also provide a reduction in the required dose of therapeutic compounds required for treatment. Furthermore, therapeutic bioavailability, pharmacodynamics, and pharmacokinetic parameters can be modulated through the use of nucleic acid bioconjugates of the invention. Non-limiting examples of such bioconjugates are described in Vargeese et al., U.S. Ser. No. 10/201,394, filed Aug. 13, 2001; and Matulic-Adamic et al., U.S. Ser. No. 60/362,016, filed Mar. 6, 2002.

Alternatively, certain siNA molecules of the instant invention can be expressed within cells from eukaryotic promoters (e.g., Izant and Weintraub, 1985, Science, 229, 345; McGarry and Lindquist, 1986, Proc. Natl. Acad. Sci., USA 83, 399; Scanlon et al., 1991, Proc. Natl. Acad. Sci. USA, 88, 10591-5; Kashani-Sabet et al., 1992, Antisense Res. Dev., 2, 3-15; Dropulic et al., 1992, J. Virol., 66, 1432-41; Weerasinghe et al., 1991, J. Virol., 65, 5531-4; Ojwang et al., 1992, Proc. Natl. Acad. Sci. USA, 89, 10802-6; Chen et al., 1992, Nucleic Acids Res., 20, 4581-9; Sarver et al., 1990 Science, 247, 1222-1225; Thompson et al., 1995, Nucleic Acids Res., 23, 2259; Good et al., 1997, Gene Therapy, 4, 45. Those skilled in the art realize that any nucleic acid can be expressed in eukaryotic cells from the appropriate DNA/RNA vector. The activity of such nucleic acids can be augmented by their release from the primary transcript by a enzymatic nucleic acid (Draper et al., PCT WO 93/23569, and Sullivan et al., PCT WO 94/02595; Ohkawa et al., 1992, Nucleic Acids Symp. Ser., 27, 15-6; Taira et al., 1991, Nucleic Acids Res., 19, 5125-30; Ventura et al., 1993, Nucleic Acids Res., 21, 3249-55; Chowrira et al., 1994, J. Biol. Chem., 269, 25856.

In another aspect of the invention, RNA molecules of the present invention can be expressed from transcription units (see for example Couture et al., 1996, TIG., 12, 510) inserted into DNA or RNA vectors. The recombinant vectors can be DNA plasmids or viral vectors. siNA expressing viral vectors can be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. In another embodiment, pol III based constructs are used to express nucleic acid molecules of the invention (see for example Thompson, U.S. Pats. Nos. 5,902,880 and 6,146,886). The recombinant vectors capable of expressing the siNA molecules can be delivered as described above, and persist in target cells. Alternatively, viral vectors can be used that provide for transient expression of nucleic acid molecules. Such vectors can be repeatedly administered as necessary. Once expressed, the siNA molecule interacts with the target mRNA and generates an RNAi response. Delivery of siNA molecule expressing vectors can be systemic, such as by intravenous or intra-muscular administration, by administration to target cells ex-planted from a subject followed by reintroduction into the subject, or by any other means that would allow for introduction into the desired target cell (for a review see Couture et al., 1996, TIG., 12, 510).

In one aspect the invention features an expression vector comprising a nucleic acid sequence encoding at least one siNA molecule of the instant invention. The expression vector can encode one or both strands of a siNA duplex, or a single self-complementary strand that self hybridizes into a siNA duplex. The nucleic acid sequences encoding the siNA molecules of the instant invention can be operably linked in a manner that allows expression of the siNA molecule (see for example Paul et al., 2002, Nature Biotechnology, 19, 505; Miyagishi and Taira, 2002, Nature Biotechnology, 19, 497; Lee et al., 2002, Nature Biotechnology, 19, 500; and Novina et al., 2002, Nature Medicine, advance online publication doi: 10.1038/nm725).

In another aspect, the invention features an expression vector comprising: a) a transcription initiation region (e.g., eukaryotic pol I, II or III initiation region); b) a transcription termination region (e.g., eukaryotic pol I, II or III termination region); and c) a nucleic acid sequence encoding at least one of the siNA molecules of the instant invention, wherein said sequence is operably linked to said initiation region and said termination region in a manner that allows expression and/or delivery of the siNA molecule. The vector can optionally include an open reading frame (ORF) for a protein operably linked on the 5′ side or the 3′-side of the sequence encoding the siNA of the invention; and/or an intron (intervening sequences).

Transcription of the siNA molecule sequences can be driven from a promoter for eukaryotic RNA polymerase I (pol I), RNA polymerase II (pol II), or RNA polymerase III (pol III). Transcripts from pol II or pol III promoters are expressed at high levels in all cells; the levels of a given pol II promoter in a given cell type depends on the nature of the gene regulatory sequences (enhancers, silencers, etc.) present nearby. Prokaryotic RNA polymerase promoters are also used, providing that the prokaryotic RNA polymerase enzyme is expressed in the appropriate cells (Elroy-Stein and Moss, 1990, Proc. Natl. Acad. Sci. USA, 87, 6743-7; Gao and Huang 1993, Nucleic Acids Res., 21, 2867-72; Lieber et al., 1993, Methods Enzymol., 217, 47-66; Zhou et al., 1990, Mol. Cell. Biol., 10, 4529-37). Several investigators have demonstrated that nucleic acid molecules expressed from such promoters can function in mammalian cells (e.g. Kashani-Sabet et al., 1992, Antisense Res. Dev., 2, 3-15; Ojwang et al., 1992, Proc. Natl. Acad. Sci. USA, 89, 10802-6; Chen et al., 1992, Nucleic Acids Res., 20, 4581-9; Yu et al., 1993, Proc. Natl. Acad. Sci. USA, 90, 6340-4; L'Huillier et al., 1992, EMBO J., 11, 4411-8; Lisziewicz et al., 1993, Proc. Natl. Acad. Sci. U.S.A., 90, 8000-4; Thompson et al., 1995, Nucleic Acids Res., 23, 2259; Sullenger & Cech, 1993, Science, 262, 1566). More specifically, transcription units such as the ones derived from genes encoding U6 small nuclear (snRNA), transfer RNA (tRNA) and adenovirus VA RNA are useful in generating high concentrations of desired RNA molecules such as siNA in cells (Thompson et al., supra; Couture and Stinchcomb, 1996, supra; Noonberg et al., 1994, Nucleic Acid Res., 22, 2830; Noonberg et al., U.S. Pat. No. 5,624,803; Good et al., 1997, Gene Ther., 4, 45; Beigelman et al., International PCT Publication No. WO 96/18736. The above siNA transcription units can be incorporated into a variety of vectors for introduction into mammalian cells, including but not restricted to, plasmid DNA vectors, viral DNA vectors (such as adenovirus or adeno-associated virus vectors), or viral RNA vectors (such as retroviral or alphavirus vectors) (for a review see Couture and Stinchcomb, 1996, supra).

In another aspect the invention features an expression vector comprising a nucleic acid sequence encoding at least one of the siNA molecules of the invention in a manner that allows expression of that siNA molecule. The expression vector comprises in one embodiment; a) a transcription initiation region; b) a transcription termination region; and c) a nucleic acid sequence encoding at least one strand of the siNA molecule, wherein the sequence is operably linked to the initiation region and the termination region in a manner that allows expression and/or delivery of the siNA molecule.

In another embodiment the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an open reading frame; and d) a nucleic acid sequence encoding at least one strand of a siNA molecule, wherein the sequence is operably linked to the 3′-end of the open reading frame and wherein the sequence is operably linked to the initiation region, the open reading frame and the termination region in a manner that allows expression and/or delivery of the siNA molecule. In yet another embodiment, the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an intron; and d) a nucleic acid sequence encoding at least one siNA molecule, wherein the sequence is operably linked to the initiation region, the intron and the termination region in a manner which allows expression and/or delivery of the nucleic acid molecule.

In another embodiment, the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an intron; d) an open reading frame; and e) a nucleic acid sequence encoding at least one strand of a siNA molecule, wherein the sequence is operably linked to the 3′-end of the open reading frame and wherein the sequence is operably linked to the initiation region, the intron, the open reading frame and the termination region in a manner which allows expression and/or delivery of the siNA molecule.

Histone Deacetylase (HDAC) Biology and Biochemistry

The following discussion is adapted from Acharya et al., 2005, Molecular Pharmacology Fast Forward, June 14, 1-49. The epigenome is defined by DNA methylation patterns and the associated post-translational modifications of histones, which are integral in gene expression. For example, this histone code determines the expression status of individual genes dependent upon their localization on the chromatin. The histone deacetylases (HDACs) play a major role in keeping the balance between the acetylated and deacetylated states of chromatin and eventually regulate gene expression by altering the dynamic balance between heterochromatin and euchromatin. Recent developments in understanding the cancer cell cycle, specifically the interplay with chromatin control and regulation, are providing opportunities for developing mechanism-based therapeutic drugs. Inhibitors of HDACs are under considerable exploration both non-clinically and in the clinic, in part due to their potential roles in reversing the silenced genes in transformed tumor cells by modulating transcriptional processes.

In eukaryotic cells, DNA has been conserved over evolution in a condensed and densely packed higher order structure generally called chromatin. Chromatin, which is present in the interphase nucleus, comprises regular repeating units of nucleosomes, which represent the principal protein-nucleic acid interface. The major components of chromatin include nucleic acids (DNA and RNA), which are negatively charged, associated proteins, including histones, that are positively charged at neutral pH, and non-histone chromosomal proteins which are acidic at neutral pH. Within the nucleus, chromatin can exist in two different forms; heterochromatin, which is highly compact and transcriptionally inactive form, or euchromatin, which is loosely packed and is accessible to RNA polymerases for involvement in transcriptional processes and resulting gene expression. A nucleosome is a complex of about 146 nucleotide base pairs of DNA wrapped around the core histone octamer that helps organize chromatin structure. The histone octamer is composed of two copies of each of H2A, H2B, H3 and H4 proteins that are very basic mainly due to positively charged amino-terminal side chains rich in the amino acid lysine. Post-translational and other changes in chromatin, such as acetylation/deacetylation at lysine residues, methylation at lysine or arginine residues, phosphorylation at serine resides, ubiquitylation at lysines, and/or ADP ribosylation, are mediated by chemical modification of various sites on the N-terminal tail. The structural modification of histones is regulated mainly by acetylation and deacetylation of the N-terminal tail and is crucial in modulating gene expression, as it affects the interaction of DNA with transcription-regulatory non-nucleosomal protein complexes.

The balance between the acetylated and deacetylated states of histones is mediated by two different sets of enzymes called histone acetyltransferases (HATs) and histone deacetylases (HDACs). HATs preferentially acetylate specific lysine substrates among other non-histone protein substrates and transcription factors, impacting DNA-binding properties and in turn, altering levels of gene transcription and ultimate gene expression. HDACs restore the positive charge on lysine residues by removing acetyl groups and are therefore involved primarily in the repression of gene transcription by condensing chromatin structure. As such, open lysine residues can attach firmly to the phosphate backbone of DNA, preventing transcription. In this tight conformation, transcription factors, regulatory complexes, and RNA polymerases cannot bind to the DNA and gene expression is effectively silenced. Acetylation relaxes the DNA conformation, making it accessible to the transcription machinery. High levels of acetylation of core histones are seen in chromatin-containing genes, which are highly transcribed genes, whereas those genes that are silent are associated with low levels of acetylation.

Because inappropriate silencing of critical genes can result in one or both hits of tumor suppressor gene (TSG) inactivation in cancer, theoretically, the reactivation of affected TSGs could have an enormous therapeutic value in preventing and treating cancer and other proliferative diseases and conditions.

The equilibrium of steady state acetylation and deacetylation is tightly controlled by the antagonistic effect of both HATs and HDACs, which in turn regulates transcription status of not just histones, but also of other substrates such as p53. Several groups of proteins with HAT activity have been identified to date, including GNAT (Gcn5-related N-acetyl transferase) family, MYST (monocytic leukemia zinc finger protein) group, TIP60 (TAT-interactive protein) and the p300/CBP (CREB-binding protein) family. HATs act as large multiprotein complexes containing other HATs, coactivators for transcription factors, and certain co-repressors. HATs, which bind non-histone protein substrates and transcription factors, have also been called factor acetyltransferases. Acetylation of these transcription factors can also affect their DNA binding properties and resulting gene transcription. HAT genes are associated with some cancers, for example, HAT genes can be overexpressed, translocated, or mutated in both hematological and epithelial cancers. The translocation of HATs, CREB-binding protein (CBP), and p300 acetyltransferases into certain genes have given rise to various hematological malignancies.

There are three major groups or classes of mammalian histone deacetylases (HDACs) based on their structural homologies to the three distinct yeast HDACs: Rpd3 (class I), Hda1 (class II), and Sir2/Hst (class III). Class III HDACs consist of the large family of sirtuins (silent information regulators or SIRs) that are evolutionarily distinct, with a unique enzymatic mechanism dependent on the cofactor NAD+, and which are all virtually unaffected by all HDAC inhibitors in current development. Both class I and class II HDACs contain an active site zinc as a critical component of their enzymatic pocket, have been extensively described to have an association with cancers, and are thought to be comparably inhibited by all HDAC inhibitors in development thus far. The Rpd3 homologous class I include HDACs 1, 2, 3 and 8, are widely expressed in various tissues and are primarily localized in the nucleus. Hda1 homologous class II HDACs 4, 5, 6, 7, 9a, 9b and 10, are much larger in size, display limited tissue distribution and can shuttle between the nucleus and cytoplasm, which suggests different functions and cellular substrates from Class I HDACs. HDACs 6 and 10 are unique as they have two catalytic domains, while HDACs 4, 8 and 9 are expressed to greater extent in tumor tissues and have been shown to be specifically involved in differentiation processes.

HDACs usually interact as constituents of large protein complexes that down-regulate genes through association with co-repressors, such as nuclear receptor corepressor (NcoR), silencing mediator for retinoid and thyroid hormone receptor (SMRT), transcription factors, estrogen receptors (ER), p53, cell-cycle specific regulators like retinoblastoma (Rb), E2F and other HDACs, as well as histones, but they can also bind to their corresponding receptor directly. Class III HDACs (sirtuins, SIR T1, 2, 3, 4, 5, 6 and 7) are generally not inhibited by class I and II HDAC inhibitors, but instead are inhibited by nicotinamide (Vitamin B3). Nicotinamide inhibits an NAD-dependent p53 deacetylation process which is induced by SIR2alpha, and also enhances p53 acetylation levels in vivo. It has been shown that by restraining mammalian forkhead proteins, specifically foxo3a, SIRT1 can also reduce apoptosis. The inhibition of forkhead activity by SIRT1 parallels the effect of this particular deacetylase on the tumor suppressor p53. These findings have significant implications regarding an important role for Sirtuins in modulating the sensitivity of cells in p53-dependent apoptotic response and the possible effect in areas ranging from cancer therapy to lifespan extension.

Chromatin modification and cancer related DNA gene expression is controlled by an assembly of nucleoproteins that includes histones and other architectural components of chromatin, non-histone DNA-bound regulators, and additional chromatin-bound polypeptides. Changes in growth and differentiation leading to transformation and malignancy appear to occur by alterations in transcriptional control and gene silencing. It has become increasingly apparent that imbalances of both DNA methylation and histone acetylation play an important role in cancer development and progression. Unlike normal cells, in cancerous cells, changes in genome expression are associated with the remodeling of long regions of regulatory DNA sequences, including promoters, enhancers, locus control regions, and insulators, into specific chromatin architecture. These specific changes in DNA architecture result in a general molecular signature for a specific type of cancer and complement its DNA methylation based component.

The changes in the infrastructure of chromatin organization over a target promoter are more profound than those observed by these enzymes acting independently. In addition to acetylation, histone tails undergo other modifications including methylation, phosphorylation, ubiquitylation and adenosine diphosphate ribosylation. Disruption of HAT and HDAC function is associated with the development of cancer and malignant cells target chromatin-remodeling pathways as a means of disrupting transcriptional regulation and control. Of the various hypotheses describing deregulation mechanisms, the following three have been put forth frequently: i) disordered hyperacetylation could activate promoters that are normally repressed leading to inappropriate expression of proteins, ii) abnormally decreased acetylation levels of promoter regions could repress the expression of genes necessary for a certain phenotype and iii) mistargeted or aberrant recruitment of HAT/HDAC activity could act as a pathological trigger for oncogenesis.

Based upon the current understanding of HAT and HDAC function, the modulation of HAT and HDAC and other related genes is instrumental in the development of new therapeutics for cancer and proliferative diseases and conditions. As such, modulation of HDACs using small interfering nucleic acid (siNA) mediated RNAi represents a novel approach to the treatment and study of diseases and conditions related to HDAC activity and/or gene expression.

EXAMPLES

The following are non-limiting examples showing the selection, isolation, synthesis and activity of nucleic acids of the instant invention.

Example 1
Tandem Synthesis of siNA Constructs

Exemplary siNA molecules of the invention are synthesized in tandem using a cleavable linker, for example, a succinyl-based linker. Tandem synthesis as described herein is followed by a one-step purification process that provides RNAi molecules in high yield. This approach is highly amenable to siNA synthesis in support of high throughput RNAi screening, and can be readily adapted to multi-column or multi-well synthesis platforms.

After completing a tandem synthesis of a siNA oligo and its complement in which the 5′-terminal dimethoxytrityl (5′-O-DMT) group remains intact (trityl on synthesis), the oligonucleotides are deprotected as described above. Following deprotection, the siNA sequence strands are allowed to spontaneously hybridize. This hybridization yields a duplex in which one strand has retained the 5′-O-DMT group while the complementary strand comprises a terminal 5′-hydroxyl. The newly formed duplex behaves as a single molecule during routine solid-phase extraction purification (Trityl-On purification) even though only one molecule has a dimethoxytrityl group. Because the strands form a stable duplex, this dimethoxytrityl group (or an equivalent group, such as other trityl groups or other hydrophobic moieties) is all that is required to purify the pair of oligos, for example, by using a C18 cartridge.

Standard phosphoramidite synthesis chemistry is used up to the point of introducing a tandem linker, such as an inverted deoxy abasic succinate or glyceryl succinate linker (see FIG. 1) or an equivalent cleavable linker. A non-limiting example of linker coupling conditions that can be used includes a hindered base such as diisopropylethylamine (DIPA) and/or DMAP in the presence of an activator reagent such as Bromotripyrrolidinophosphoniumhexaflurorophosphate (PyBrOP). After the linker is coupled, standard synthesis chemistry is utilized to complete synthesis of the second sequence leaving the terminal the 5′-O-DMT intact. Following synthesis, the resulting oligonucleotide is deprotected according to the procedures described herein and quenched with a suitable buffer, for example with 50 mM NaOAc or 1.5M NH₄H₂CO₃.

Purification of the siNA duplex can be readily accomplished using solid phase extraction, for example, using a Waters C18 SepPak 1 g cartridge conditioned with 1 column volume (CV) of acetonitrile, 2 CV H2O, and 2 CV 50 mM NaOAc. The sample is loaded and then washed with 1 CV H2O or 50 mM NaOAc. Failure sequences are eluted with 1 CV 14% ACN (Aqueous with 50 mM NaOAc and 50 mM NaCl). The column is then washed, for example with 1 CV H2O followed by on-column detritylation, for example by passing 1 CV of 1% aqueous trifluoroacetic acid (TFA) over the column, then adding a second CV of 1% aqueous TFA to the column and allowing to stand for approximately 10 minutes. The remaining TFA solution is removed and the column washed with H20 followed by 1 CV 1M NaCl and additional H2O. The siNA duplex product is then eluted, for example, using 1 CV 20% aqueous CAN.

FIG. 2 provides an example of MALDI-TOF mass spectrometry analysis of a purified siNA construct in which each peak corresponds to the calculated mass of an individual siNA strand of the siNA duplex. The same purified siNA provides three peaks when analyzed by capillary gel electrophoresis (CGE), one peak presumably corresponding to the duplex siNA, and two peaks presumably corresponding to the separate siNA sequence strands. Ion exchange HPLC analysis of the same siNA contract only shows a single peak. Testing of the purified siNA construct using a luciferase reporter assay described below demonstrated the same RNAi activity compared to siNA constructs generated from separately synthesized oligonucleotide sequence strands.

Example 2
Identification of Potential siNA Target Sites in any RNA Sequence

The sequence of an RNA target of interest, such as a HDAC mRNA transcript, is screened for target sites, for example by using a computer folding algorithm. In a non-limiting example, the sequence of a HDAC gene or HDAC RNA gene transcript derived from a database, such as Genbank, is used to generate siNA targets having complementarity to the target. Such sequences can be obtained from a database, or can be determined experimentally as known in the art. Target sites that are known, for example, those target sites determined to be effective target sites based on studies with other nucleic acid molecules, for example ribozymes or antisense, or those targets known to be associated with a disease, trait, or condition such as those sites containing mutations or deletions, can be used to design siNA molecules targeting those sites. Various parameters can be used to determine which sites are the most suitable target sites within the target RNA sequence. These parameters include but are not limited to secondary or tertiary RNA structure, the nucleotide base composition of the target sequence, the degree of homology between various regions of the target sequence, or the relative position of the target sequence within the RNA transcript. Based on these determinations, any number of target sites within the RNA transcript can be chosen to screen siNA molecules for efficacy, for example by using in vitro RNA cleavage assays, cell culture, or animal models. In a non-limiting example, anywhere from 1 to 1000 target sites are chosen within the transcript based on the size of the siNA construct to be used. High throughput screening assays can be developed for screening siNA molecules using methods known in the art, such as with multi-well or multi-plate assays to determine efficient reduction in target gene expression.

Example 3
Selection of siNA Molecule Target Sites in a RNA

The following non-limiting steps can be used to carry out the selection of siNAs targeting a given gene sequence or transcript.

1. The target sequence is parsed in silico into a list of all fragments or subsequences of a particular length, for example 23 nucleotide fragments, contained within the target sequence. This step is typically carried out using a custom Perl script, but commercial sequence analysis programs such as Oligo, MacVector, or the GCG Wisconsin Package can be employed as well.

2. In some instances the siNAs correspond to more than one target sequence; such would be the case for example in targeting different transcripts of the same gene, targeting different transcripts of more than one gene, or for targeting both the human gene and an animal homolog. In this case, a subsequence list of a particular length is generated for each of the targets, and then the lists are compared to find matching sequences in each list. The subsequences are then ranked according to the number of target sequences that contain the given subsequence; the goal is to find subsequences that are present in most or all of the target sequences. Alternately, the ranking can identify subsequences that are unique to a target sequence, such as a mutant target sequence. Such an approach would enable the use of siNA to target specifically the mutant sequence and not effect the expression of the normal sequence.

3. In some instances the siNA subsequences are absent in one or more sequences while present in the desired target sequence; such would be the case if the siNA targets a gene with a paralogous family member that is to remain untargeted. As in case 2 above, a subsequence list of a particular length is generated for each of the targets, and then the lists are compared to find sequences that are present in the target gene but are absent in the untargeted paralog.

4. The ranked siNA subsequences can be further analyzed and ranked according to GC content. A preference can be given to sites containing 30-70% GC, with a further preference to sites containing 40-60% GC.

5. The ranked siNA subsequences can be further analyzed and ranked according to self-folding and internal hairpins. Weaker internal folds are preferred; strong hairpin structures are to be avoided.

6. The ranked siNA subsequences can be further analyzed and ranked according to whether they have runs of GGG or CCC in the sequence. GGG (or even more Gs) in either strand can make oligonucleotide synthesis problematic and can potentially interfere with RNAi activity, so it is avoided whenever better sequences are available. CCC is searched in the target strand because that will place GGG in the antisense strand.

7. The ranked siNA subsequences can be further analyzed and ranked according to whether they have the dinucleotide UU (uridine dinucleotide) on the 3′-end of the sequence, and/or AA on the 5′-end of the sequence (to yield 3′ UU on the antisense sequence). These sequences allow one to design siNA molecules with terminal TT thymidine dinucleotides.

8. Four or five target sites are chosen from the ranked list of subsequences as described above. For example, in subsequences having 23 nucleotides, the right 21 nucleotides of each chosen 23-mer subsequence are then designed and synthesized for the upper (sense) strand of the siNA duplex, while the reverse complement of the left 21 nucleotides of each chosen 23-mer subsequence are then designed and synthesized for the lower (antisense) strand of the siNA duplex (see Table II). If terminal TT residues are desired for the sequence (as described in paragraph 7), then the two 3′ terminal nucleotides of both the sense and antisense strands are replaced by TT prior to synthesizing the oligos.

9. The siNA molecules are screened in an in vitro, cell culture or animal model system to identify the most active siNA molecule or the most preferred target site within the target RNA sequence.

10. Other design considerations can be used when selecting target nucleic acid sequences, see, for example, Reynolds et al., 2004, Nature Biotechnology, 22, 326-330 and Ui-Tei et al., 2004, Nucleic Acids Research, 32, 936-948.

In an alternate approach, a pool of siNA constructs specific to a target sequence is used to screen for target sites in cells expressing target RNA, such as cultured Jurkat, HeLa, A549 or 293T cells. The general strategy used in this approach is shown in FIG. 9. Cells expressing the target RNA are transfected with the pool of siNA constructs and cells that demonstrate a phenotype associated with target inhibition are sorted. The pool of siNA constructs can be expressed from transcription cassettes inserted into appropriate vectors (see for example FIG. 7 and FIG. 8). The siNA from cells demonstrating a positive phenotypic change (e.g., decreased proliferation, decreased target mRNA levels or decreased target protein expression), are sequenced to determine the most suitable target site(s) within the target target RNA sequence.

Example 4
siNA Design

siNA target sites were chosen by analyzing sequences of HDAC target RNA sequences using the parameters described in Example 3 above and optionally prioritizing the target sites on the basis of folding (structure of any given sequence analyzed to determine siNA accessibility to the target). Such sites can also be chosen by using a library of siNA molecules as described in Example 3, or alternately by using an in vitro siNA system as described in Example 6 herein. siNA molecules were designed that could bind each target and are optionally individually analyzed by computer folding to assess whether the siNA molecule can interact with the target sequence. Chemical modification criteria were applied in designing chemically modified siNA molecules (see for example Table III) based on stabilization chemistry motifs described herein (see for example Table IV). Varying the length of the siNA molecules can be chosen to optimize activity. Generally, a sufficient number of complementary nucleotide bases are chosen to bind to, or otherwise interact with, the target RNA, but the degree of complementarity can be modulated to accommodate siNA duplexes or varying length or base composition. By using such methodologies, siNA molecules can be designed to target sites within any known RNA sequence, for example those RNA sequences corresponding to the any gene transcript.

Chemically modified siNA constructs are designed to provide nuclease stability for systemic administration in vivo and/or improved pharmacokinetic, localization, and delivery properties while preserving the ability to mediate RNAi activity. Chemical modifications as described herein are introduced synthetically using synthetic methods described herein and those generally known in the art. The synthetic siNA constructs are then assayed for nuclease stability in serum and/or cellular/tissue extracts (e.g. liver extracts). The synthetic siNA constructs are also tested in parallel for RNAi activity using an appropriate assay, such as a luciferase reporter assay as described herein or another suitable assay that can quantity RNAi activity. Synthetic siNA constructs that possess both nuclease stability and RNAi activity can be further modified and re-evaluated in stability and activity assays. The chemical modifications of the stabilized active siNA constructs can then be applied to any siNA sequence targeting any chosen RNA and used, for example, in target screening assays to pick lead siNA compounds for therapeutic development (see for example FIG. 11).

Example 5
Chemical Synthesis and Purification of siNA

siNA molecules can be designed to interact with various sites in the RNA message, for example, target sequences within the RNA sequences described herein. The sequence of one strand of the siNA molecule(s) is complementary to the target site sequences described above. The siNA molecules can be chemically synthesized using methods described herein. Inactive siNA molecules that are used as control sequences can be synthesized by scrambling the sequence of the siNA molecules such that it is not complementary to the target sequence. Generally, siNA constructs can by synthesized using solid phase oligonucleotide synthesis methods as described herein (see for example Usman et al., U.S. Pat. Nos. 5,804,683; 5,831,071; 5,998,203; 6,117,657; 6,353,098; 6,362,323; 6,437,117; 6,469,158; Scaringe et al., U.S. Pat. Nos. 6,111,086; 6,008,400; 6,111,086 all incorporated by reference herein in their entirety).

In a non-limiting example, RNA oligonucleotides are synthesized in a stepwise fashion using the phosphoramidite chemistry as is known in the art. Standard phosphoramidite chemistry involves the use of nucleosides comprising any of 5′-O-dimethoxytrityl, 2′-O-tert-butyldimethylsilyl, 3′-O-2-Cyanoethyl N,N-diisopropylphosphoroamidite groups, and exocyclic amine protecting groups (e.g. N6-benzoyl adenosine, N4 acetyl cytidine, and N2-isobutyryl guanosine). Alternately, 2′-O-Silyl Ethers can be used in conjunction with acid-labile 2′-O-orthoester protecting groups in the synthesis of RNA as described by Scaringe supra. Differing 2′ chemistries can require different protecting groups, for example 2′-deoxy-2′-amino nucleosides can utilize N-phthaloyl protection as described by Usman et al., U.S. Pat. No. 5,631,360, incorporated by reference herein in its entirety).

During solid phase synthesis, each nucleotide is added sequentially (3′- to 5′-direction) to the solid support-bound oligonucleotide. The first nucleoside at the 3′-end of the chain is covalently attached to a solid support (e.g., controlled pore glass or polystyrene) using various linkers. The nucleotide precursor, a ribonucleoside phosphoramidite, and activator are combined resulting in the coupling of the second nucleoside phosphoramidite onto the 5′-end of the first nucleoside. The support is then washed and any unreacted 5′-hydroxyl groups are capped with a capping reagent such as acetic anhydride to yield inactive 5′-acetyl moieties. The trivalent phosphorus linkage is then oxidized to a more stable phosphate linkage. At the end of the nucleotide addition cycle, the 5′-O-protecting group is cleaved under suitable conditions (e.g., acidic conditions for trityl-based groups and Fluoride for silyl-based groups). The cycle is repeated for each subsequent nucleotide.

Modification of synthesis conditions can be used to optimize coupling efficiency, for example by using differing coupling times, differing reagent/phosphoramidite concentrations, differing contact times, differing solid supports and solid support linker chemistries depending on the particular chemical composition of the siNA to be synthesized. Deprotection and purification of the siNA can be performed as is generally described in Usman et al., U.S. Pat. No. 5,831,071, U.S. Pat. No. 6,353,098, U.S. Pat. No. 6,437,117, and Bellon et al., U.S. Pat. No. 6,054,576, U.S. Pat. No. 6,162,909, U.S. Pat. No. 6,303,773, or Scaringe supra, incorporated by reference herein in their entireties. Additionally, deprotection conditions can be modified to provide the best possible yield and purity of siNA constructs. For example, applicant has observed that oligonucleotides comprising 2′-deoxy-2′-fluoro nucleotides can degrade under inappropriate deprotection conditions. Such oligonucleotides are deprotected using aqueous methylamine at about 35° C. for 30 minutes. If the 2′-deoxy-2′-fluoro containing oligonucleotide also comprises ribonucleotides, after deprotection with aqueous methylamine at about 35° C. for 30 minutes, TEA-HF is added and the reaction maintained at about 65° C. for an additional 15 minutes.

Example 6
RNAi In Vitro Assay to Assess siNA Activity

An in vitro assay that recapitulates RNAi in a cell-free system is used to evaluate siNA constructs targeting HDAC RNA targets. The assay comprises the system described by Tuschl et al., 1999, Genes and Development, 13, 3191-3197 and Zamore et al., 2000, Cell, 101, 25-33 adapted for use with a target RNA. A Drosophila extract derived from syncytial blastoderm is used to reconstitute RNAi activity in vitro. Target RNA is generated via in vitro transcription from an appropriate target expressing plasmid using T7 RNA polymerase or via chemical synthesis as described herein. Sense and antisense siNA strands (for example 20 uM each) are annealed by incubation in buffer (such as 100 mM potassium acetate, 30 mM HEPES-KOH, pH 7.4, 2 mM magnesium acetate) for 1 minute at 90° C. followed by 1 hour at 37° C., then diluted in lysis buffer (for example 100 mM potassium acetate, 30 mM HEPES-KOH at pH 7.4, 2 mM magnesium acetate). Annealing can be monitored by gel electrophoresis on an agarose gel in TBE buffer and stained with ethidium bromide. The Drosophila lysate is prepared using zero to two-hour-old embryos from Oregon R flies collected on yeasted molasses agar that are dechorionated and lysed. The lysate is centrifuged and the supernatant isolated. The assay comprises a reaction mixture containing 50% lysate [vol/vol], RNA (10-50 pM final concentration), and 10% [vol/vol] lysis buffer containing siNA (10 nM final concentration). The reaction mixture also contains 10 mM creatine phosphate, 10 ug/ml creatine phosphokinase, 100 um GTP, 100 uM UTP, 100 uM CTP, 500 uM ATP, 5 mM DTT, 0.1 U/uL RNasin (Promega), and 100 uM of each amino acid. The final concentration of potassium acetate is adjusted to 100 mM. The reactions are pre-assembled on ice and preincubated at 25° C. for 10 minutes before adding RNA, then incubated at 25° C. for an additional 60 minutes. Reactions are quenched with 4 volumes of 1.25×Passive Lysis Buffer (Promega). Target RNA cleavage is assayed by RT-PCR analysis or other methods known in the art and are compared to control reactions in which siNA is omitted from the reaction.

Alternately, internally-labeled target RNA for the assay is prepared by in vitro transcription in the presence of [alpha-³²P] CTP, passed over a G50 Sephadex column by spin chromatography and used as target RNA without further purification. Optionally, target RNA is 5′-³²P-end labeled using T4 polynucleotide kinase enzyme. Assays are performed as described above and target RNA and the specific RNA cleavage products generated by RNAi are visualized on an autoradiograph of a gel. The percentage of cleavage is determined by PHOSPHOR IMAGER® (autoradiography) quantitation of bands representing intact control RNA or RNA from control reactions without siNA and the cleavage products generated by the assay.

In one embodiment, this assay is used to determine target sites in the HDAC RNA target for siNA mediated RNAi cleavage, wherein a plurality of siNA constructs are screened for RNAi mediated cleavage of the HDAC RNA target, for example, by analyzing the assay reaction by electrophoresis of labeled target RNA, or by northern blotting, as well as by other methodology well known in the art.

Example 7
Nucleic Acid Inhibition of HDAC RNA In Vivo

siNA molecules targeted to the human HDAC RNA are designed and synthesized as described above. These nucleic acid molecules can be tested for cleavage activity in vivo, for example, using the following procedure.

Two formats are used to test the efficacy of siNAs against a given target (e.g., HDAC 11). First, the reagents are tested in cell culture using, for example, Jurkat, HeLa, A549 or 293T cells, to determine the extent of RNA and protein inhibition. siNA reagents are selected against the target HDAC 11 RNA as described herein. RNA inhibition is measured after delivery of these reagents by a suitable transfection agent to, for example, Jurkat, HeLa, A549 or 293T cells. Relative amounts of target RNA are measured versus actin using real-time PCR monitoring of amplification (eg., ABI 7700 TAQMAN®). A comparison is made to a mixture of oligonucleotide sequences made to unrelated targets or to a randomized siNA control with the same overall length and chemistry, but randomly substituted at each position. Primary and secondary lead reagents are chosen for the target and optimization performed. After an optimal transfection agent concentration is chosen, a RNA time-course of inhibition is performed with the lead siNA molecule. In addition, a cell-plating format can be used to determine RNA inhibition.

Delivery of siNA to Cells

Cells (e.g., Jurkat, HeLa, A549 or 293T cells) are seeded, for example, at 1×10⁵cells per well of a six-well dish in EGM-2 (BioWhittaker) the day before transfection. siNA (final concentration, for example 20 nM) and cationic lipid (e.g., final concentration 2 μg/ml) are complexed in EGM basal media (Biowhittaker) at 37° C. for 30 minutes in polystyrene tubes. Following vortexing, the complexed siNA is added to each well and incubated for the times indicated. For initial optimization experiments, cells are seeded, for example, at 1×13 in 96 well plates and siNA complex added as described. Efficiency of delivery of siNA to cells is determined using a fluorescent siNA complexed with lipid. Cells in 6-well dishes are incubated with siNA for 24 hours, rinsed with PBS and fixed in 2% paraformaldehyde for 15 minutes at room temperature. Uptake of siNA is visualized using a fluorescent microscope.

TAQMAN® (Real-Time PCR Monitoring of Amplification) and Lightcycler Quantification of mRNA

Total RNA is prepared from cells following siNA delivery, for example, using Qiagen RNA purification kits for 6-well or Rneasy extraction kits for 96-well assays. For TAQMAN® analysis (real-time PCR monitoring of amplification), dual-labeled probes are synthesized with the reporter dye, FAM or JOE, covalently linked at the 5′-end and the quencher dye TAMRA conjugated to the 3′-end. One-step RT-PCR amplifications are performed on, for example, an ABI PRISM 7700 Sequence Detector using 50 μl reactions consisting of 10 μl total RNA, 100 nM forward primer, 900 nM reverse primer, 100 nM probe, 1×TaqMan PCR reaction buffer (PE-Applied Biosystems), 5.5 mM MgCl₂, 300 μM each dATP, dCTP, dGTP, and dTTP, 10 U RNase Inhibitor (Promega), 1.25 U AMPLITAQ GOLD® (DNA polymerase) (PE-Applied Biosystems) and 10 U M-MLV Reverse Transcriptase (Promega). The thermal cycling conditions can consist of 30 minutes at 48° C., 10 minutes at 95° C., followed by 40 cycles of 15 seconds at 95° C. and 1 minute at 60° C. Quantitation of mRNA levels is determined relative to standards generated from serially diluted total cellular RNA (300, 100, 33, 11 ng/reaction) and normalizing to β-actin or GAPDH mRNA in parallel TAQMAN® reactions (real-time PCR monitoring of amplification). For each gene of interest an upper and lower primer and a fluorescently labeled probe are designed. Real time incorporation of SYBR Green I dye into a specific PCR product can be measured in glass capillary tubes using a lightcyler. A standard curve is generated for each primer pair using control cRNA. Values are represented as relative expression to GAPDH in each sample.

Western Blotting

Nuclear extracts can be prepared using a standard micro preparation technique (see for example Andrews and Faller, 1991, Nucleic Acids Research, 19, 2499). Protein extracts from supernatants are prepared, for example using TCA precipitation. An equal volume of 20% TCA is added to the cell supernatant, incubated on ice for 1 hour and pelleted by centrifugation for 5 minutes. Pellets are washed in acetone, dried and resuspended in water. Cellular protein extracts are run on a 10% Bis-Tris NuPage (nuclear extracts) or 4-12% Tris-Glycine (supernatant extracts) polyacrylamide gel and transferred onto nitro-cellulose membranes. Non-specific binding can be blocked by incubation, for example, with 5% non-fat milk for 1 hour followed by primary antibody for 16 hour at 4° C. Following washes, the secondary antibody is applied, for example (1:10,000 dilution) for 1 hour at room temperature and the signal detected with SuperSignal reagent (Pierce).

Example 8
Models Useful to Evaluate the Down-Regulation of Target Gene Expression

Evaluating the efficacy of siNA molecules of the invention in animal models is an important prerequisite to human clinical trials. Various animal models of cancer, proliferative, ocular, etc. diseases, conditions, or disorders as are known in the art can be adapted for use for pre-clinical evaluation of the efficacy of nucleic acid compositions of the invetention in modulating target gene expression toward therapeutic, cosmetic, or research use. Non-limiting examples of pre-models useful in evaluating HDAC inhibitory compounds for therapeutic use can be found in Acharya et al., 2005, Molecular Pharmacology Fast Forward, June 14, 1-49; Curtin and Glaser, 2003, Curr. Med. Chem., 10, 2372-92; and Filocamo et al., International PCT Publication No. WO 05/071079, all incorporated by reference herein.

Example 9
RNAi Mediated Inhibition of Target Gene Expression

In Vitro siNA Mediated Inhibition of HDAC RNA

siNA constructs are tested for efficacy in reducing target RNA expression in cells, (e.g., HEKn/HEKa, HeLa, A549, A375 cells). Cells are plated approximately 24 hours before transfection in 96-well plates at 5,000-7,500 cells/well, 100 μl/well, such that at the time of transfection cells are 70-90% confluent. For transfection, annealed siNAs are mixed with the transfection reagent (e.g., Lipofectamine 2000, Invitrogen) in a volume of 50 μl/well and incubated for 20 minutes at room temperature. The siNA transfection mixtures are added to cells to give a final siNA concentration of 25 nM in a volume of 150 μl. Each siNA transfection mixture is added to 3 wells for triplicate siNA treatments. Cells are incubated at 37° for 24 hours in the continued presence of the siNA transfection mixture. At 24 hours, RNA is prepared from each well of treated cells. The supernatants with the transfection mixtures are first removed and discarded, then the cells are lysed and RNA prepared from each well. Target gene expression following treatment is evaluated by RT-PCR for the target gene and for a control gene (36B4, an RNA polymerase subunit) for normalization. The triplicate data is averaged and the standard deviations determined for each treatment. Normalized data are graphed and the percent reduction of target mRNA by active siNAs in comparison to their respective inverted control siNAs is determined.

Example 10
Indications

Particular conditions and disease states that are associated with HDAC gene expression modulation using siNA molecules of the invention include, but are not limited to cancer, proliferative, ocular, allograft rejection and age related diseases, conditions, or disorders as described herein or otherwise known in the art, and any other diseases, conditions or disorders that are related to or will respond to the levels of a HDAC (e.g., HDAC target protein or target polynucleotide) in a cell or tissue, alone or in combination with other therapies.

Example 11
Multifunctional siNA Inhibition of Target RNA Expression

Multifunctional siNA Design

Once target sites have been identified for multifunctional siNA constructs, each strand of the siNA is designed with a complementary region of length, for example, of about 18 to about 28 nucleotides, that is complementary to a different target nucleic acid sequence. Each complementary region is designed with an adjacent flanking region of about 4 to about 22 nucleotides that is not complementary to the target sequence, but which comprises complementarity to the complementary region of the other sequence (see for example FIG. 16). Hairpin constructs can likewise be designed (see for example FIG. 17). Identification of complementary, palindrome or repeat sequences that are shared between the different target nucleic acid sequences can be used to shorten the overall length of the multifunctional siNA constructs (see for example FIGS. 18 and 19).

In a non-limiting example, three additional categories of additional multifunctional siNA designs are presented that allow a single siNA molecule to silence multiple targets. The first method utilizes linkers to join siNAs (or multiunctional siNAs) in a direct manner. This can allow the most potent siNAs to be joined without creating a long, continuous stretch of RNA that has potential to trigger an interferon response. The second method is a dendrimeric extension of the overlapping or the linked multifunctional design; or alternatively the organization of siNA in a supramolecular format. The third method uses helix lengths greater than 30 base pairs. Processing of these siNAs by Dicer will reveal new, active 5′ antisense ends. Therefore, the long siNAs can target the sites defined by the original 5′ ends and those defined by the new ends that are created by Dicer processing. When used in combination with traditional multifunctional siNAs (where the sense and antisense strands each define a target) the approach can be used for example to target 4 or more sites.

I. Tethered Bifunctional siNAs

The basic idea is a novel approach to the design of multifunctional siNAs in which two antisense siNA strands are annealed to a single sense strand. The sense strand oligonucleotide contains a linker (e.g., non-nulcoetide linker as described herein) and two segments that anneal to the antisense siNA strands (see FIG. 22). The linkers can also optionally comprise nucleotide-based linkers. Several potential advantages and variations to this approach include, but are not limited to:

1. The two antisense siNAs are independent. Therefore, the choice of target sites is not constrained by a requirement for sequence conservation between two sites. Any two highly active siNAs can be combined to form a multifunctional siNA.
2. When used in combination with target sites having homology, siNAs that target a sequence present in two genes (e.g., different isoforms), the design can be used to target more than two sites. A single multifunctional siNA can be, for example, used to target RNA of two different target RNAs.
3. Multifunctional siNAs that use both the sense and antisense strands to target a gene can also be incorporated into a tethered multifuctional design. This leaves open the possibility of targeting 6 or more sites with a single complex.
4. It can be possible to anneal more than two antisense strand siNAs to a single tethered sense strand.
5. The design avoids long continuous stretches of dsRNA. Therefore, it is less likely to initiate an interferon response.
6. The linker (or modifications attached to it, such as conjugates described herein) can improve the pharmacokinetic properties of the complex or improve its incorporation into liposomes. Modifications introduced to the linker should not impact siNA activity to the same extent that they would if directly attached to the siNA (see for example FIGS. 27 and 28).
7. The sense strand can extend beyond the annealed antisense strands to provide additional sites for the attachment of conjugates.
8. The polarity of the complex can be switched such that both of the antisense 3′ ends are adjacent to the linker and the 5′ ends are distal to the linker or combination thereof.

Dendrimer and Supramolecular siNAs

In the dendrimer siNA approach, the synthesis of siNA is initiated by first synthesizing the dendrimer template followed by attaching various functional siNAs. Various constructs are depicted in FIG. 23. The number of functional siNAs that can be attached is only limited by the dimensions of the dendrimer used.

Supramolecular Approach to Multifunctional siNA

The supramolecular format simplifies the challenges of dendrimer synthesis. In this format, the siNA strands are synthesized by standard RNA chemistry, followed by annealing of various complementary strands. The individual strand synthesis contains an antisense sense sequence of one siNA at the 5′-end followed by a nucleic acid or synthetic linker, such as hexaethyleneglyol, which in turn is followed by sense strand of another siNA in 5′ to 3′ direction. Thus, the synthesis of siNA strands can be carried out in a standard 3′ to 5′ direction. Representative examples of trifunctional and tetrafunctional siNAs are depicted in FIG. 24. Based on a similar principle, higher functionality siNA constucts can be designed as long as efficient annealing of various strands is achieved.

Dicer Enabled Multifunctional siNA

Using bioinformatic analysis of multiple targets, stretches of identical sequences shared between differeing target sequences can be identified ranging from about two to about fourteen nucleotides in length. These identical regions can be designed into extended siNA helixes (e.g., >30 base pairs) such that the processing by Dicer reveals a secondary functional 5′-antisense site (see for example FIG. 25). For example, when the first 17 nucleotides of a siNA antisense strand (e.g., 21 nucleotide strands in a duplex with 3′-TT overhangs) are complementary to a target RNA, robust silencing was observed at 25 nM. 80% silencing was observed with only 16 nucleotide complementarity in the same format.

Incorporation of this property into the designs of siNAs of about 30 to 40 or more base pairs results in additional multifunctional siNA constructs. The example in FIG. 25 illustrates how a 30 base-pair duplex can target three distinct sequences after processing by Dicer-RNaseIII; these sequences can be on the same mRNA or separate RNAs, such as viral and host factor messages, or multiple points along a given pathway (e.g., inflammatory cascades). Furthermore, a 40 base-pair duplex can combine a bifunctional design in tandem, to provide a single duplex targeting four target sequences. An even more extensive approach can include use of homologous sequences to enable five or six targets silenced for one multifunctional duplex. The example in FIG. 25 demonstrates how this can be achieved. A 30 base pair duplex is cleaved by Dicer into 22 and 8 base pair products from either end (8 b.p. fragments not shown). For ease of presentation the overhangs generated by dicer are not shown—but can be compensated for. Three targeting sequences are shown. The required sequence identity overlapped is indicated by grey boxes. The N's of the parent 30 b.p. siNA are suggested sites of 2′-OH positions to enable Dicer cleavage if this is tested in stabilized chemistries. Note that processing of a 30mer duplex by Dicer RNase III does not give a precise 22+8 cleavage, but rather produces a series of closely related products (with 22+8 being the primary site). Therefore, processing by Dicer will yield a series of active siNAs. Another non-limiting example is shown in FIG. 26. A 40 base pair duplex is cleaved by Dicer into 20 base pair products from either end. For ease of presentation the overhangs generated by dicer are not shown—but can be compensated for. Four targeting sequences are shown in four colors, blue, light-blue and red and orange. The required sequence identity overlapped is indicated by grey boxes. This design format can be extended to larger RNAs. If chemically stabilized siNAs are bound by Dicer, then strategically located ribonucleotide linkages can enable designer cleavage products that permit our more extensive repertoire of multiifunctional designs. For example cleavage products not limited to the Dicer standard of approximately 22-nucleotides can allow multifunctional siNA constructs with a target sequence identity overlap ranging from, for example, about 3 to about 15 nucleotides.

Example 12
Diagnostic Uses

The siNA molecules of the invention can be used in a variety of diagnostic applications, such as in the identification of molecular targets (e.g., RNA) in a variety of applications, for example, in clinical, industrial, environmental, agricultural and/or research settings. Such diagnostic use of siNA molecules involves utilizing reconstituted RNAi systems, for example, using cellular lysates or partially purified cellular lysates. siNA molecules of this invention can be used as diagnostic tools to examine genetic drift and mutations within diseased cells or to detect the presence of endogenous or exogenous, for example viral, RNA in a cell. The close relationship between siNA activity and the structure of the target RNA allows the detection of mutations in any region of the molecule, which alters the base-pairing and three-dimensional structure of the target RNA. By using multiple siNA molecules described in this invention, one can map nucleotide changes, which are important to RNA structure and function in vitro, as well as in cells and tissues. Cleavage of target RNAs with siNA molecules can be used to inhibit gene expression and define the role of specified gene products in the progression of disease or infection. In this manner, other genetic targets can be defined as important mediators of the disease. These experiments will lead to better treatment of the disease progression by affording the possibility of combination therapies (e.g., multiple siNA molecules targeted to different genes, siNA molecules coupled with known small molecule inhibitors, or intermittent treatment with combinations siNA molecules and/or other chemical or biological molecules). Other in vitro uses of siNA molecules of this invention are well known in the art, and include detection of the presence of mRNAs associated with a disease, infection, or related condition. Such RNA is detected by determining the presence of a cleavage product after treatment with a siNA using standard methodologies, for example, fluorescence resonance emission transfer (FRET).

In a specific example, siNA molecules that cleave only wild-type or mutant forms of the target RNA are used for the assay. The first siNA molecules (i.e., those that cleave only wild-type forms of target RNA) are used to identify wild-type RNA present in the sample and the second siNA molecules (i.e., those that cleave only mutant forms of target RNA) are used to identify mutant RNA in the sample. As reaction controls, synthetic substrates of both wild-type and mutant RNA are cleaved by both siNA molecules to demonstrate the relative siNA efficiencies in the reactions and the absence of cleavage of the “non-targeted” RNA species. The cleavage products from the synthetic substrates also serve to generate size markers for the analysis of wild-type and mutant RNAs in the sample population. Thus, each analysis requires two siNA molecules, two substrates and one unknown sample, which is combined into six reactions. The presence of cleavage products is determined using an RNase protection assay so that full-length and cleavage fragments of each RNA can be analyzed in one lane of a polyacrylamide gel. It is not absolutely required to quantify the results to gain insight into the expression of mutant RNAs and putative risk of the desired phenotypic changes in target cells. The expression of mRNA whose protein product is implicated in the development of the phenotype (i.e., disease related or infection related) is adequate to establish risk. If probes of comparable specific activity are used for both transcripts, then a qualitative comparison of RNA levels is adequate and decreases the cost of the initial diagnosis. Higher mutant form to wild-type ratios are correlated with higher risk whether RNA levels are compared qualitatively or quantitatively.

All patents and publications mentioned in the specification are indicative of the levels of skill of those skilled in the art to which the invention pertains. All references cited in this disclosure are incorporated by reference to the same extent as if each reference had been incorporated by reference in its entirety individually.

One skilled in the art would readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The methods and compositions described herein as presently representative of preferred embodiments are exemplary and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those skilled in the art, which are encompassed within the spirit of the invention, are defined by the scope of the claims.

It will be readily apparent to one skilled in the art that varying substitutions and modifications can be made to the invention disclosed herein without departing from the scope and spirit of the invention. Thus, such additional embodiments are within the scope of the present invention and the following claims. The present invention teaches one skilled in the art to test various combinations and/or substitutions of chemical modifications described herein toward generating nucleic acid constructs with improved activity for mediating RNAi activity. Such improved activity can comprise improved stability, improved bioavailability, and/or improved activation of cellular responses mediating RNAi. Therefore, the specific embodiments described herein are not limiting and one skilled in the art can readily appreciate that specific combinations of the modifications described herein can be tested without undue experimentation toward identifying siNA molecules with improved RNAi activity.

The invention illustratively described herein suitably can be practiced in the absence of any element or elements, limitation or limitations that are not specifically disclosed herein. Thus, for example, in each instance herein any of the terms “comprising”, “consisting essentially of”, and “consisting of” may be replaced with either of the other two terms. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments, optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the description and the appended claims.

In addition, where features or aspects of the invention are described in terms of Markush groups or other grouping of alternatives, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group or other group.

TABLE IHDAC Accession NumbersNM_004964Homo sapiens histone deacetylase 1 (HDAC1), mRNANM_001527Homo sapiens histone deacetylase 2 (HDAC2), mRNANM_024665Homo sapiens nuclear receptor co-repressor/HDAC3 complex subunit (FLJ12894),mRNANM_003883Homo sapiens histone deacetylase 3 (HDAC3), mRNANM_006037Homo sapiens histone deacetylase 4 (HDAC4), mRNANM_005474Homo sapiens histone deacetylase 5 (HDAC5), mRNANM_139205Homo sapiens histone deacetylase 5 (HDAC5), transcript variant 2, mRNANM_006044Homo sapiens histone deacetylase 6 (HDAC6), mRNANM_016596Homo sapiens histone deacetylase 7A (HDAC7A), transcript variant 2, mRNANM_015401Homo sapiens histone deacetylase 7A (HDAC7A), transcript variant 1, mRNANM_018486Homo sapiens histone deacetylase 8 (HDAC8), mRNANM_058177Homo sapiens histone deacetylase 9 (HDAC9-PENDING), transcript variant 2,mRNANM_058176Homo sapiens histone deacetylase 9 (HDAC9-PENDING), transcript variant 1,mRNANM_014707Homo sapiens histone deacetylase 9 (HDAC9-PENDING), transcript variant 3,mRNANM_178423Homo sapiens histone deacetylase 9 (HDAC9), transcript variant 4, mRNANM_178425Homo sapiens histone deacetylase 9 (HDAC9), transcript variant 5, mRNANM_032019Homo sapiens histone deacetylase 10 (HDAC10), mRNANM_024827Homo sapiens histone deacetylase 11 (HDAC11), mRNANM_012238Homo sapiens sirtuin (silent mating type information regulation 2 homolog) 1 (S. cerevisiae)(SIRT1), mRNANM_012237Homo sapiens sirtuin (silent mating type information regulation 2 homolog) 2 (S. cerevisiae)(SIRT2), transcript variant 1, mRNANM_030593Homo sapiens sirtuin (silent mating type information regulation 2 homolog) 2 (S. cerevisiae)(SIRT2), transcript variant 2, mRNANM_012239Homo sapiens sirtuin (silent mating type information regulation 2 homolog) 3 (S. cerevisiae)(SIRT3), mRNANM_012240Homo sapiens sirtuin (silent mating type information regulation 2 homolog) 4 (S. cerevisiae)(SIRT4), mRNANM_012241Homo sapiens sirtuin (silent mating type information regulation 2 homolog) 5 (S. cerevisiae)(SIRT5), transcript variant 1, mRNANM_031244Homo sapiens sirtuin (silent mating type information regulation 2 homolog) 5 (S. cerevisiae)(SIRT5), transcript variant 2, mRNAXM_372781Homo sapiens similar to NAD-dependent deacetylase sirtuin 5 (SIR2-like protein 5)(LOC391047), mRNANM_016539Homo sapiens sirtuin (silent mating type information regulation 2 homolog) 6 (S. cerevisiae)(SIRT6), mRNANM_016538Homo sapiens sirtuin (silent mating type information regulation 2 homolog) 7 (S. cerevisiae)(SIRT7), mRNA

TABLE II

HDAC siNA and Target Sequences

Seq

Seq

Seq

Pos
Seq
ID
UPos
Upper seq
ID
LPos
Lower seq
ID

HDAC1: NM_004964.2

3
GCGGAGCCGCGGGCGGGAG
1
3
GCGGAGCCGCGGGCGGGAG
1
21
CUCCCGCCCGCGGCUCCGC
116

21
GGGCGGACGGACCGACUGA
2
21
GGGCGGACGGACCGACUGA
2
39
UCAGUCGGUCCGUCCGCCC
117

39
ACGGUAGGGACGGGAGGCG
3
39
ACGGUAGGGACGGGAGGCG
3
57
CGCCUCCCGUCCCUACCGU
118

57
GAGCAAGAUGGCGCAGACG
4
57
GAGCAAGAUGGCGCAGACG
4
75
CGUCUGCGCCAUCUUGCUC
119

75
GCAGGGCACCCGGAGGAAA
5
75
GCAGGGCACCCGGAGGAAA
5
93
UUUCCUCCGGGUGCCCUGC
120

93
AGUCUGUUACUACUACGAC
6
93
AGUCUGUUACUACUACGAC
6
111
GUCGUAGUAGUAACAGACU
121

111
CGGGGAUGUUGGAAAUUAC
7
111
CGGGGAUGUUGGAAAUUAC
7
129
GUAAUUUCCAACAUCCCCG
122

129
CUAUUAUGGACAAGGCCAC
8
129
CUAUUAUGGACAAGGCCAC
8
147
GUGGCCUUGUCCAUAAUAG
123

147
CCCAAUGAAGCCUCACCGA
9
147
CCCAAUGAAGCCUCACCGA
9
165
UCGGUGAGGCUUCAUUGGG
124

165
AAUCCGCAUGACUCAUAAU
10
165
AAUCCGCAUGACUCAUAAU
10
183
AUUAUGAGUCAUGCGGAUU
125

183
UUUGGUGCUCAACUAUGGU
11
183
UUUGCUGCUCAAGUAUGGU
11
201
ACCAUAGUUGAGCAGCAAA
126

201
UCUCUACCGAAAAAUGGAA
12
201
UCUCUACCGAAAAAUGGAA
12
219
UUCCAUUUUUCGGUAGAGA
127

219
AAUCUAUCGCCCUCACAAA
13
219
AAUCUAUCGCCCUCACAAA
13
237
UUUGUGAGGGCGAUAGAUU
128

237
AGCCAAUGCUGAGGAGAUG
14
237
AGCCAAUGCUGAGGAGAUG
14
255
CAUCUCCUCAGCAUUGGCU
129

255
GACCAAGUACCACAGCGAU
15
255
GACCAAGUACCACAGCGAU
15
273
AUCGCUGUGGUACUUGGUC
130

273
UGACUACAUUAAAUUCUUG
16
273
UGACUACAUUAAAUUCUUG
16
291
CAAGAAUUUAAUGUAGUCA
131

291
GCGCUCCAUCCGUCCAGAU
17
291
GCGCUCCAUCCGUCCAGAU
17
309
AUCUGGACGGAUGGAGCGC
132

309
UAACAUGUCGGAGUACAGC
18
309
UAACAUGUCGGAGUACAGC
18
327
GGUGUACUCCGACAUGUUA
133

327
CAAGCAGAUGCAGAGAUUC
19
327
CAAGCAGAUGCAGAGAUUC
19
345
GAAUCUCUGCAUCUGCUUG
134

345
CAACGUUGGUGAGGACUGU
20
345
CAACGUUGGUGAGGACUGU
20
363
ACAGUCCUCACCAACGUUG
135

363
UCCAGUAUUCGAUGGCCUG
21
363
UCCAGUAUUCGAUGGCCUG
21
381
CAGGCCAUCGAAUACUGGA
136

381
GUUUGAGUUCUGUCAGUUG
22
381
GUUUGAGUUCUGUCAGUUG
22
399
CAACUGACAGAACUCAAAC
137

399
GUCUACUGGUGGUUCUGUG
23
399
GUCUACUGGUGGUUCUGUG
23
417
CACAGAACCACCAGUAGAC
138

417
GGCAAGUGCUGUGAAACUU
24
417
GGCAAGUGCUGUGAAACUU
24
435
AAGUUUCACAGCACUUGCC
139

435
UAAUAAGCAGCAGACGGAC
25
435
UAAUAAGCAGCAGACGGAC
25
453
GUCCGUCUGCUGCUUAUUA
140

453
CAUCGCUGUGAAUUGGGCU
26
453
CAUCGCUGUGAAUUGGGCU
26
471
AGCCCAAUUCACAGCGAUG
141

471
UGGGGGCCUGCACCAUGCA
27
471
UGGGGGCCUGCACCAUGCA
27
489
UGCAUGGUGCAGGCCCCCA
142

489
AAAGAAGUCCGAGGCAUCU
28
489
AAAGAAGUCCGAGGCAUCU
28
507
AGAUGCCUCGGACUUCUUU
143

507
UGGCUUCUGUUACGUCAAU
29
507
UGGCUUCUGUUACGUCAAU
29
525
AUUGACGUAACAGAAGCCA
144

525
UGAUAUCGUCUUGGCCAUC
30
525
UGAUAUCGUCUUGGCCAUC
30
543
GAUGGCCAAGACGAUAUCA
145

543
CCUGGAACUGCUAAAGUAU
31
543
CCUGGAACUGCUAAAGUAU
31
561
AUACUUUAGCAGUUCCAGG
146

561
UCACCAGAGGGUGCUGUAC
32
561
UCACCAGAGGGUGCUGUAC
32
579
GUACAGGACCCUCUGGUGA
147

579
CAUUGACAUUGAUAUUCAC
33
579
CAUUGACAUUGAUAUUCAC
33
597
GUGAAUAUCAAUGUCAAUG
148

597
CCAUGGUGACGGCGUGGAA
34
597
CCAUGGUGACGGCGUGGAA
34
615
UUCCACGCCGUCACCAUGG
149

615
AGAGGCCUUCUACACCACG
35
615
AGAGGCCUUCUACACCACG
35
633
CGUGGUGUAGAAGGCCUCU
150

633
GGACCGGGUCAUGACUGUG
36
633
GGACCGGGUCAUGACUGUG
36
651
CACAGUCAUGACCCGGUCC
151

651
GUCCUUUCAUAAGUAUGGA
37
651
GUCCUUUCAUAAGUAUGGA
37
669
UCCAUACUUAUGAAAGGAC
152

669
AGAGUACUUCCCAGGAACU
38
669
AGAGUACUUCCCAGGAACU
38
687
AGUUCCUGGGAAGUACUCU
153

687
UGGGGACCUACGGGAUAUC
39
687
UGGGGACCUACGGGAUAUC
39
705
GAUAUCCCGUAGGUCCCCA
154

705
CGGGGCUGGCAAAGGCAAG
40
705
CGGGGCUGGCAAAGGCAAG
40
723
CUUGCCUUUGCCAGCCCCG
155

723
GUAUUAUGCUGUUAACUAC
41
723
GUAUUAUGCUGUUAACUAC
41
741
GUAGUUAACAGCAUAAUAC
156

741
CCCGCUCCGAGACGGGAUU
42
741
CCCGCUCCGAGACGGGAUU
42
759
AAUCCCGUCUCGGAGCGGG
157

759
UGAUGACGAGUCCUAUGAG
43
759
UGAUGACGAGUCCUAUGAG
43
777
CUCAUAGGACUCGUCAUCA
158

777
GGCCAUUUUCAAGCCGGUC
44
777
GGCCAUUUUCAAGCCGGUC
44
795
GACCGGCUUGAAAAUGGCC
159

795
CAUGUCCAAAGUAAUGGAG
45
795
CAUGUCCAAAGUAAUGGAG
45
813
CUCCAUUACUUUGGACAUG
160

813
GAUGUUCCAGCCUAGUGCG
46
813
GAUGUUCCAGCCUAGUGCG
46
831
CGCACUAGGCUGGAACAUC
161

831
GGUGGUCUUACAGUGUGGC
47
831
GGUGGUCUUACAGUGUGGC
47
849
GCCACACUGUAAGACCACC
162

849
CUCAGACUCCCUAUCUGGG
48
849
CUCAGACUCCCUAUCUGGG
48
867
CCCAGAUAGGGAGUCUGAG
163

867
GGAUCGGUUAGGUUGCUUC
49
867
GGAUCGGUUAGGUUGCUUC
49
885
GAAGCAACCUAACCGAUCC
164

885
CAAUCUAACUAUCAAAGGA
50
885
CAAUCUAACUAUCPAAGGA
50
903
UCCUUUGAUAGUUAGAUUG
165

903
ACACGCCAAGUGUGUGGAA
51
903
ACACGCCAAGUGUGUGGAA
51
921
UUCCACACACUUGGCGUGU
166

921
AUUUGUCAAGAGCUUUAAC
52
921
AUUUGUCAAGAGCUUUAAC
52
939
GUUAAAGCUCUUGACAAAU
167

939
CCUGCCUAUGCUGAUGCUG
53
939
CCUGCCUAUGCUGAUGCUG
53
957
CAGCAUCAGCAUAGGCAGG
168

957
GGGAGGCGGUGGUUACACC
54
957
GGGAGGCGGUGGUUACACC
54
975
GGUGUAACCACCGCCUCCC
169

975
CAUUCGUAACGUUGCCCGG
55
975
CAUUCGUAACGUUGCCCGG
55
993
CCGGGCAACGUUACGAAUG
170

993
GUGCUGGACAUAUGAGACA
56
993
GUGCUGGACAUAUGAGACA
56
1011
UGUCUCAUAUGUCCAGCAC
171

1011
AGCUGUGGCCCUGGAUACG
57
1011
AGCUGUGGCCCUGGAUACG
57
1029
CGUAUCCAGGGCCACAGCU
172

1029
GGAGAUCCCUAAUGAGCUU
58
1029
GGAGAUCCCUAAUGAGCUU
58
1047
AAGCUCAUUAGGGAUCUCC
173

1047
UCCAUACAAUGACUACUUU
59
1047
UCCAUACAAUGACUACUUU
59
1065
AAAGUAGUCAUUGUAUGGA
174

1065
UGAAUACUUUGGACCAGAU
60
1065
UGAAUACUUUGGACCAGAU
60
1083
AUCUGGUCCAAAGUAUUCA
175

1083
UUUCAAGCUCCACAUCAGU
61
1083
UUUCAAGCUCCACAUCAGU
61
1101
ACUGAUGUGGAGCUUGAAA
176

1101
UCCUUCCAAUAUGACUAAC
62
1101
UCCUUCCAAUAUGACUAAC
62
1119
GUUAGUCAUAUUGGAAGGA
177

1119
CCAGAACACGAAUGAGUAC
63
1119
CCAGAACACGAAUGAGUAC
63
1137
GUACUCAUUCGUGUUCUGG
178

1137
CCUGGAGAAGAUCAAACAG
64
1137
CCUGGAGAAGAUCAAACAG
64
1155
CUGUUUGAUCUUCUCCAGG
179

1155
GCGACUGUUUGAGAACCUU
65
1155
GCGACUGUUUGAGAACCUU
65
1173
AAGGUUCUCAAACAGUCGC
180

1173
UAGAAUGCUGCCGCACGCA
66
1173
UAGAAUGCUGCCGCACGCA
66
1191
UGCGUGCGGCAGCAUUCUA
181

1191
ACCUGGGGUCCAAAUGCAG
67
1191
ACCUGGGGUCCAAAUGCAG
67
1209
CUGCAUUUGGACCCCAGGU
182

1209
GGCGAUUCCUGAGGACGCC
68
1209
GGCGAUUCCUGAGGACGCC
68
1227
GGCGUCCUCAGGAAUCGCC
183

1227
CAUCCCUGAGGAGAGUGGC
69
1227
CAUCCCUGAGGAGAGUGGC
69
1245
GCCACUCUCCUCAGGGAUG
184

1245
CGAUGAGGACGAAGACGAC
70
1245
CGAUGAGGACGAAGAGGAC
70
1263
GUCGUCUUCGUCCUCAUCG
185

1263
CCCUGACAAGCGCAUCUCG
71
1263
CCCUGACAAGCGCAUCUCG
71
1281
CGAGAUGCGCUUGUCAGGG
186

1281
GAUCUGCUCCUCUGACAAA
72
1281
GAUCUGCUCCUCUGACAAA
72
1299
UUUGUCAGAGGAGCAGAUC
187

1299
ACGAAUUGCCUGUGAGGAA
73
1299
ACGAAUUGCCUGUGAGGAA
73
1317
UUCCUCACAGGCAAUUCGU
188

1317
AGAGUUCUCCGAUUCUGAA
74
1317
AGAGUUCUCCGAUUCUGAA
74
1335
UUCAGAAUCGGAGAACUCU
189

1335
AGAGGAGGGAGAGGGGGGC
75
1335
AGAGGAGGGAGAGGGGGGC
75
1353
GCCCCCCUCUCCCUCCUCU
190

1353
CCGCAAGAACUCUUCCAAC
76
1353
CCGCAAGAACUCUUCCAAC
76
1371
GUUGGAAGAGUUCUUGCGG
191

1371
CUUCAAAAAAGCCAAGAGA
77
1371
CUUCAAAAAAGCCAAGAGA
77
1389
UCUCUUGGCUUUUUUGAAG
192

1389
AGUCAAAACAGAGGAUGAA
78
1389
AGUCAAAACAGAGGAUGAA
78
1407
UUCAUCCUCUGUUUUGACU
193

1407
AAAAGAGAAAGACCCAGAG
79
1407
AAAAGAGAAAGACCCAGAG
79
1425
CUCUGGGUCUUUCUCUUUU
194

1425
GGAGAAGAAAGAAGUCACC
80
1425
GGAGAAGAAAGAAGUCACC
80
1443
GGUGACUUCUUUCUUCUCC
195

1443
CGAAGAGGAGAAAACCAAG
81
1443
CGAAGAGGAGAAAACCAAG
81
1461
CUUGGUUUUCUCCUCUUCG
196

1461
GGAGGAGAAGCCAGAAGCC
82
1461
GGAGGAGAAGCCAGAAGCC
82
1479
GGCUUCUGGCUUCUCCUCC
197

1479
CAAAGGGGUCAAGGAGGAG
83
1479
CAAAGGGGUCAAGGAGGAG
83
1497
CUCCUCCUUGACCCCUUUG
198

1497
GGUCAAGUUGGCCUGAAUG
84
1497
GGUCAAGUUGGCCUGAAUG
84
1515
CAUUCAGGCCAACUUGACC
199

1515
GGACCUCUCCAGCUCUGGC
85
1515
GGACCUCUCCAGCUCUGGC
85
1533
GCCAGAGCUGGAGAGGUCC
200

1533
CUUCCUGCUGAGUCCCUCA
86
1533
CUUCCUGCUGAGUCCCUCA
86
1551
UGAGGGACUCAGCAGGAAG
201

1551
ACGUUUCUUCCCCAACCCC
87
1551
ACGUUUCUUCCCCAACCCC
87
1569
GGGGUUGGGGAAGAAACGU
202

1569
CUCAGAUUUUAUAUUUUCU
88
1569
CUCAGAUUUUAUAUUUUCU
88
1587
AGAAAAUAUAAAAUCUGAG
203

1587
UAUUUCUCUGUGUAUUUAU
89
1587
UAUUUCUCUGUGUAUUUAU
89
1605
AUAAAUACACAGAGAAAUA
204

1605
UAUAAAAAUUUAUUAAAUA
90
1605
UAUAAAAAUUUAUUAAAUA
90
1623
UAUUUAAUAAAUUUUUAUA
205

1623
AUAAAUAUCCCCAGGGACA
91
1623
AUAAAUAUCCCCAGGGACA
91
1641
UGUCCCUGGGGAUAUUUAU
206

1641
AGAAACCAAGGCCCCGAGC
92
1641
AGAAACCAAGGCCCCGAGC
92
1659
GCUCGGGGCCUUGGUUUCU
207

1659
CUCAGGGCAGCUGUGCUGG
93
1659
CUCAGGGCAGCUGUGCUGG
93
1677
CCAGCACAGCUGCCCUGAG
208

1677
GGUGAGCUCUUCCAGGAGC
94
1677
GGUGAGCUCUUCCAGGAGC
94
1695
GCUCCUGGAAGAGCUCACC
209

1695
CCACCUUGCCACCCAUUCU
95
1695
CCACCUUGCCACCCAUUCU
95
1713
AGAAUGGGUGGCAAGGUGG
210

1713
UUCCCGUUCUUAACUUUGA
96
1713
UUCCCGUUCUUAACUUUGA
96
1731
UCAAAGUUAAGAACGGGAA
211

1731
AACCAUAAAGGGUGCCAGG
97
1731
AACCAUAAAGGGUGCCAGG
97
1749
CCUGGCACCCUUUAUGGUU
212

1749
GUCUGGGUGAAAGGGAUAC
98
1749
GUCUGGGUGAAAGGGAUAC
98
1767
GUAUCCCUUUGACCCAGAC
213

1767
CUUUUAUGCAACCAUAAGA
99
1767
CUUUUAUGCAACCAUAAGA
99
1785
UCUUAUGGUUGCAUAAAAG
214

1785
ACAAACUCCUGAAAUGCCA
100
1785
ACAAACUCCUGAAAUGCCA
100
1803
UGGCAUUUCAGGAGUUUGU
215

1803
AAGUGCCUGCUUAGUAGCU
101
1803
AAGUGCCUGCUUAGUAGCU
101
1821
AGCUACUAAGCAGGCACUU
216

1821
UUUGGAAAGGUGCCCUUAU
102
1821
UUUGGAAAGGUGCCCUUAU
102
1839
AUAAGGGCACCUUUCCAAA
217

1839
UUGAACAUUCUAGAAGGGG
103
1839
UUGAACAUUCUAGAAGGGG
103
1857
CCCCUUCUAGAAUGUUCAA
218

1857
GUGGCUGGGUCUUCAAGGA
104
1857
GUGGCUGGGUCUUCAAGGA
104
1875
UCCUUGAAGACCCAGCCAC
219

1875
AUCUCCUGUUUUUUUCAGG
105
1875
AUCUCCUGUUUUUUUCAGG
105
1893
CCUGAAAAAAACAGGAGAU
220

1893
GCUCCUAAAGUAACAUCAG
106
1893
GCUCCUAAAGUAACAUCAG
106
1911
CUGAUGUUACUUUAGGAGC
221

1911
GCCAUUUUUAGAUUGGUUC
107
1911
GCCAUUUUUAGAUUGGUUC
107
1929
GAACCAAUCUAAAAAUGGC
222

1929
CUGUUUUCGUACCUUCCCA
108
1929
CUGUUUUCGUACCUUCCCA
108
1947
UGGGAAGGUACGAAAACAG
223

1947
ACUGGCCUCAAGUGAGCCA
109
1947
ACUGGCCUCAAGUGAGCCA
109
1965
UGGCUCACUUGAGGCCAGU
224

1965
AAGAAACACUGCCUGCCCU
110
1965
AAGAAACACUGCCUGCCCU
110
1983
AGGGCAGGCAGUGUUUCUU
225

1983
UCUGUCUGUCUUCUCCUAA
111
1983
UCUGUCUGUCUUCUCCUAA
111
2001
UUAGGAGAAGACAGACAGA
226

2001
AUUCUGCAGGUGGAGGUUG
112
2001
AUUCUGCAGGUGGAGGUUG
112
2019
CAACCUCCACCUGCAGAAU
227

2019
GCUAGUCUAGUUUCCUUUU
113
2019
GCUAGUCUAGUUUCCUUUU
113
2037
AAAAGGAAACUAGACUAGC
228

2037
UUGAGAUACUAUUUUCAUU
114
2037
UUGAGAUACUAUUUUCAUU
114
2055
AAUGAAAAUAGUAUCUCAA
229

2055
UUUUGUGAGCCUCUUUGUA
115
2055
UUUUGUGAGCCUCUUUGUA
115
2073
UACAAAGAGGCUCACAAAA
230

HDAC2: NM_001527.1

3
CCGAGCUUUCGGCACCUCU
343
3
CCGAGCUUUCGGCACCUCU
343
21
AGAGGUGCCGAAAGCUCGG
453

21
UGCCGGGUGGUACCGAGCC
344
21
UGCCGGGUGGUACCGAGCC
344
39
GGCUCGGUACCACCCGGCA
454

39
CUUCCCGGCGCCCCCUCCU
345
39
CUUCCCGGCGCCCCCUCCU
345
57
AGGAGGGGGCGCCGGGAAG
455

57
UCUCCUCCCACCGGCCUGC
346
57
UCUCCUCCCACCGGCCUGC
346
75
GCAGGCCGGUGGGAGGAGA
456

75
CCCUUCCCCGCGGGACUAU
347
75
CCCUUCCCCGCGGGACUAU
347
93
AUAGUCCCGCGGGGAAGGG
457

93
UCGCCCCCACGUUUCCCUC
348
93
UCGCCCCCACGUUUCCCUC
348
111
GAGGGAAACGUGGGGGCGA
458

111
CAGCCCUUUUCUCUCCCGG
349
111
CAGCCCUUUUCUCUCCCGG
349
129
CCGGGAGAGAAAAGGGCUG
459

129
GCCGAGCCGCGGCGGCAGC
350
129
GCCGAGCCGCGGCGGCAGC
350
147
GCUGCCGCCGCGGCUCGGC
460

147
CAGCAGCAGCAGCAGCAGC
351
147
CAGCAGCAGCAGCAGCAGC
351
165
GCUGCUGCUGCUGCUGCUG
461

165
CAGGAGGAGGAGCCCGGUG
352
165
CAGGAGGAGGAGCCCGGUG
352
183
CACCGGGCUCCUCCUCCUG
462

183
GGCGGCGGUGGCCGGGGAG
353
183
GGCGGCGGUGGCCGGGGAG
353
201
CUCCCCGGCCACCGCCGCC
463

201
GGCCAUGGCGUACAGUCAA
354
201
GCCCAUGGCGUACAGUCAA
354
219
UUGACUGUACGCCAUGGGC
464

219
AGGAGGCGGCAAAAAAAAA
355
219
AGGAGGCGGCAAAAAAAAA
355
237
UUUUUUUUUGCCGCCUCCU
465

237
AGUCUGCUACUACUACGAC
356
237
AGUCUGCUACUACUACGAC
356
255
GUCGUAGUAGUAGCAGACU
466

255
CGGUGAUAUUGGAAAUUAU
357
255
CGGUGAUAUUGGAAAUUAU
357
273
AUAAUUUCCAAUAUCACCG
467

273
UUAUUAUGGACAGGGUCAU
358
273
UUAUUAUGGACAGGGUCAU
358
291
AUGACCCUGUCCAUAAUAA
468

291
UCCCAUGAAGCCUCAUAGA
359
291
UCCCAUGAAGCCUCAUAGA
359
309
UCUAUGAGGCUUCAUGGGA
469

309
AAUCCGCAUGACCCAUAAC
360
309
AAUCCGCAUGACCCAUAAC
360
327
GUUAUGGGUCAUGCGGAUU
470

327
CUUGCUGUUAAAUUAUGGC
361
327
CUUGCUGUUAAAUUAUGGC
361
345
GCCAUAAUUUAACAGCAAG
471

345
CUUAUACAGAAAAAUGGAA
362
345
CUUAUACAGAAAAAUGGAA
362
363
UUCCAUUUUUCUGUAUAAG
472

363
AAUAUAUAGGCCCCAUAAA
363
363
AAUAUAUAGGCCCCAUAAA
363
381
UUUAUGGGGCCUAUAUAUU
473

381
AGCCACUGCCGAAGAAAUG
364
381
AGCCACUGCCGAAGAAAUG
364
399
CAUUUCUUCGGCAGUGGCU
474

399
GACAAAAUAUCACAGUGAU
365
399
GACAAAAUAUCACAGUGAU
365
417
AUCACUGUGAUAUUUUGUC
475

417
UGAGUAUAUCAAAUUUCUA
366
417
UGAGUAUAUCAAAUUUCUA
366
435
UAGAAAUUUGAUAUACUCA
476

435
ACGGUCAAUAAGACCAGAU
367
435
ACGGUCAAUAAGACCAGAU
367
453
AUCUGGUCUUAUUGACCGU
477

453
UAACAUGUCUGAGUAUAGU
368
453
UAACAUGUCUGAGUAUAGU
368
471
ACUAUACUCAGACAUGUUA
478

471
UAAGCAGAUGCAUAUAUUU
369
471
UAAGCAGAUGCAUAUAUUU
369
489
AAAUAUAUGCAUCUGCUUA
479

489
UAAUGUUGGAGAAGAUUGU
370
489
UAAUGUUGGAGAAGAUUGU
370
507
ACAAUCUUCUCCAACAUUA
480

507
UCCAGCGUUUGAUGGACUC
371
507
UCCAGCGUUUGAUGGACUC
371
525
GAGUCCAUCAAACGCUGGA
481

525
CUUUGAGUUUUGUCAGCUC
372
525
CUUUGAGUUUUGUCAGCUC
372
543
GAGCUGACAAAACUCAAAG
482

543
CUCAACUGGCGGUUCAGUU
373
543
CUCAACUGGCGGUUCAGUU
373
561
AACUGAACCGCCAGUUGAG
483

561
UGCUGGAGCUGUGAAGUUA
374
561
UGCUGGAGCUGUGAAGUUA
374
579
UAACUUCACAGCUCCAGCA
484

579
AAACCGACAACAGACUGAU
375
579
AAACCGACAACAGACUGAU
375
597
AUCAGUCUGUUGUCGGUUU
485

597
UAUGGCUGUUAAUUGGGCU
376
597
UAUGGCUGUUAAUUGGGCU
376
615
AGCCCAAUUAACAGCCAUA
486

615
UGGAGGAUUACAUCAUGCU
377
615
UGGAGGAUUACAUCAUGCU
377
633
AGCAUGAUGUAAUCCUCCA
487

633
UAAGAAAUACGAAGCAUCA
378
633
UAAGAAAUACGAAGCAUCA
378
651
UGAUGCUUCGUAUUUCUUA
488

651
AGGAUUCUGUUACGUUAAU
379
651
AGGAUUCUGUUACGUUAAU
379
669
AUUAACGUAACAGAAUCCU
489

669
UGAUAUUGUGCUUGCCAUC
380
669
UGAUAUUGUGCUUGCCAUC
380
687
GAUGGCAAGCACAAUAUCA
490

687
CCUUGAAUUACUAAAGUAU
381
687
CCUUGAAUUACUAAAGUAU
381
705
AUACUUUAGUAAUUCAAGG
491

705
UCAUCAGAGAGUCUUAUAU
382
705
UCAUCAGAGAGUCUUAUAU
382
723
AUAUAAGACUCUCUGAUGA
492

723
UAUUGAUAUAGAUAUUCAU
383
723
UAUUGAUAUAGAUAUUCAU
383
741
AUGAAUAUCUAUAUCAAUA
493

741
UCAUGGUGAUGGUGUUGAA
384
741
UCAUGGUGAUGGUGUUGAA
384
759
UUCAACACCAUCACCAUGA
494

759
AGAAGCUUUUUAUACAACA
385
759
AGAAGCUUUUUAUACAACA
385
777
UGUUGUAUAAAAAGCUUCU
495

777
AGAUCGUGUAAUGACGGUA
386
777
AGAUCGUGUAAUGACGGUA
386
795
UACCGUCAUUACACGAUCU
496

795
AUCAUUCCAUAAAUAUGGG
387
795
AUCAUUCCAUAAAUAUGGG
387
813
CCCAUAUUUAUGGAAUGAU
497

813
GGAAUACUUUCCUGGCAGA
388
813
GGAAUACUUUCCUGGCACA
388
831
UGUGCCAGGAAAGUAUUCC
498

831
AGGAGACUUGAGGGAUAUU
389
831
AGGAGACUUGAGGGAUAUU
389
849
AAUAUCCCUCAAGUCUCCU
499

849
UGGUGCUGGAAAAGGCAAA
390
849
UGGUGCUGGAAAAGGCAAA
390
867
UUUGCCUUUUCCAGCACCA
500

867
AUACUAUGCUGUCAAUUUU
391
867
AUACUAUGCUGUCAAUUUU
391
885
AAAAUUGACAGCAUAGUAU
501

885
UCCAAUGUGUGAUGGUAUA
392
885
UCCAAUGUGUGAUGGUAUA
392
903
UAUACCAUCACACAUUGGA
502

903
AGAUGAUGAGUCAUAUGGG
393
903
AGAUGAUGAGUCAUAUGGG
393
921
CCCAUAUGACUCAUCAUCU
503

921
GCAGAUAUUUAAGCCUAUU
394
921
GCAGAUAUUUAAGCCUAUU
394
939
AAUAGGCUUAAAUAUCUGC
504

939
UAUCUCAAAGGUGAUGGAG
395
939
UAUCUCAAAGGUGAUGGAG
395
957
CUCCAUCACCUUUGAGAUA
505

957
GAUGUAUCAACCUAGUGCU
396
957
GAUGUAUCAACCUAGUGCU
396
975
AGCACUAGGUUGAUACAUC
506

975
UGUGGUAUUACAGUGUGGU
397
975
UGUGGUAUUACAGUGUGGU
397
993
ACCACACUGUAAUACCACA
507

993
UGCAGACUCAUUAUCUGGU
398
993
UGCAGACUCAUUAUCUGGU
398
1011
ACCAGAUAAUGAGUCUGCA
508

1011
UGAUAGACUGGGUUGUUUC
399
1011
UGAUAGACUGGGUUGUUUC
399
1029
GAAACAACCCAGUCUAUCA
509

1029
CAAUCUAACAGUCAAAGGU
400
1029
CAAUCUAACAGUCAAAGGU
400
1047
ACCUUUGACUGUUAGAUUG
510

1047
UCAUGCUAAAUGUGUAGAA
401
1047
UCAUGCUAAAUGUGUAGAA
401
1065
UUCUACACAUUUAGCAUGA
511

1065
AGUUGUAAAAACUUUUAAC
402
1065
AGUUGUAAAAACUUUUAAC
402
1083
GUUAAAAGUUUUUACAACU
512

1083
CUUACCAUUACUGAUGCUU
403
1083
CUUACCAUUACUGAUGCUU
403
1101
AAGCAUCAGUAAUGGUAAG
513

1101
UGGAGGAGGUGGCUACACA
404
1101
UGGAGGAGGUGGCUACACA
404
1119
UGUGUAGCCACCUCCUCCA
514

1119
AAUCCGUAAUGUUGCUCGA
405
1119
AAUCCGUAAUGUUGCUCGA
405
1137
UCGAGCAACAUUACGGAUU
515

1137
AUGUUGGACAUAUGAGACU
406
1137
AUGUUGGACAUAUGAGACU
406
1155
AGUCUCAUAUGUCCAACAU
516

1155
UGCAGUUGCCCUUGAUUGU
407
1155
UGCAGUUGCCCUUGAUUGU
407
1173
ACAAUCAAGGGCAACUGCA
517

1173
UGAGAUUCCCAAUGAGUUG
408
1173
UGAGAUUCCCAAUGAGUUG
408
1191
CAACUCAUUGGGAAUCUCA
518

1191
GCCAUAUAAUGAUUACUUU
409
1191
GCCAUAUAAUGAUUACUUU
409
1209
AAAGUAAUCAUUAUAUGGC
519

1209
UGAGUAUUUUGGACCAGAC
410
1209
UGAGUAUUUUGGACCAGAC
410
1227
GUCUGGUCCAAAAUACUCA
520

1227
CUUCAAACUGCAUAUUAGU
411
1227
CUUCAAACUGCAUAUUAGU
411
1245
ACUAAUAUGCAGUUUGAAG
521

1245
UCCUUCAAACAUGACAAAC
412
1245
UCCUUCAAACAUGACAAAC
412
1263
GUUUGUCAUGUUUGAAGGA
522

1263
CCAGAACACUCCAGAAUAU
413
1263
CCAGAACACUCCAGAAUAU
413
1281
AUAUUCUGGAGUGUUCUGG
523

1281
UAUGGAAAAGAUAAAACAG
414
1281
UAUGGAAAAGAUAAAACAG
414
1299
CUGUUUUAUCUUUUCCAUA
524

1299
GCGUUUGUUUGAAAAUUUG
415
1299
GCGUUUGUUUGAAAAUUUG
415
1317
CAAAUUUUCAAACAAACGC
525

1317
GCGCAUGUUACCUCAUGCA
416
1317
GCGCAUGUUACCUCAUGCA
416
1335
UGCAUGAGGUAACAUGCGC
526

1335
ACCUGGUGUCCAGAUGCAA
417
1335
ACCUGGUGUCCAGAUGCAA
417
1353
UUGCAUCUGGACACCAGGU
527

1353
AGCUAUUCCAGAAGAUGCU
418
1353
AGCUAUUCCAGAAGAUGCU
418
1371
AGCAUCUUCUGGAAUAGCU
528

1371
UGUUCAUGAAGACAGUGGA
419
1371
UGUUCAUGAAGACAGUGGA
419
1389
UCCACUGUCUUCAUGAACA
529

1389
AGAUGAAGAUGGAGAAGAU
420
1389
AGAUGAAGAUGGAGAAGAU
420
1407
AUCUUCUCCAUCUUCAUCU
530

1407
UCCAGACAAGAGAAUUUCU
421
1407
UCCAGACAAGAGAAUUUCU
421
1425
AGAAAUUCUCUUGUCUGGA
531

1425
UAUUCGAGCAUCAGACAAG
422
1425
UAUUCGAGCAUCAGACAAG
422
1443
CUUGUCUGAUGCUCGAAUA
532

1443
GCGGAUAGCUUGUGAUGAA
423
1443
GCGGAUAGCUUGUGAUGAA
423
1461
UUCAUCACAAGCUAUCCGC
533

1461
AGAAUUCUCAGAUUCUGAG
424
1461
AGAAUUCUCAGAUUCUGAG
424
1479
CUCAGAAUCUGAGAAUUCU
534

1479
GGAUGAAGGAGAAGGAGGU
425
1479
GGAUGAAGGAGAAGGAGGU
425
1497
ACCUCCUUCUCCUUCAUCC
535

1497
UCGAAGAAAUGUGGCUGAU
426
1497
UCGAAGAAAUGUGGCUGAU
426
1515
AUCAGCCACAUUUCUUCGA
536

1515
UCAUAAGAAAGGAGCAAAG
427
1515
UCAUAAGAAAGGAGCAAAG
427
1533
CUUUGCUCCUUUCUUAUGA
537

1533
GAAAGCUAGAAUUGAAGAA
428
1533
GAAAGCUAGAAUUGAAGAA
428
1551
UUCUUCAAUUCUAGCUUUC
538

1551
AGAUAAGAAAGAAACAGAG
429
1551
AGAUAAGAAAGAAACAGAG
429
1569
CUCUGUUUCUUUCUUAUCU
539

1569
GGACAAAAAAACAGACGUU
430
1569
GGACAAAAAAACAGACGUU
430
1587
AACGUCUGUUUUUUUGUCC
540

1587
UAAGGAAGAAGAUAAAUCC
431
1587
UAAGGAAGAAGAUAAAUCC
431
1605
GGAUUUAUCUUCUUCCUUA
541

1605
CAAGGACAACAGUGGUGAA
432
1605
CAAGGACAACAGUGGUGAA
432
1623
UUCACCACUGUUGUCCUUG
542

1623
AAAAACAGAUACCAAAGGA
433
1623
AAAAACAGAUACCAAAGGA
433
1641
UCCUUUGGUAUCUGUUUUU
543

1641
AACCAAAUCAGAACAGCUC
434
1641
AACCAAAUCAGAACAGCUC
434
1659
GAGCUGUUCUGAUUUGGUU
544

1659
CAGCAACCCCUGAAUUUGA
435
1659
CAGCAACCCCUGAAUUUGA
435
1677
UCAAAUUCAGGGGUUGCUG
545

1677
ACAGUCUCACCAAUUUCAG
436
1677
ACAGUCUCACCAAUUUCAG
436
1695
CUGAAAUUGGUGAGACUGU
546

1695
GAAAAUCAUUAAAAAGAAA
437
1695
GAAAAUCAUUAAAAAGAAA
437
1713
UUUCUUUUUAAUGAUUUUC
547

1713
AAUAUUGAAAGGAAAAUGU
438
1713
AAUAUUGAAAGGAAAAUGU
438
1731
ACAUUUUCCUUUCAAUAUU
548

1731
UUUUCUUUUUGAAGACUUC
439
1731
UUUUCUUUUUGAAGACUUC
439
1749
GAAGUCUUCAAAAAGAAAA
549

1749
CUGGCUUCAUUUUAUACUA
440
1749
CUGGCUUCAUUUUAUACUA
440
1767
UAGUAUAAAAUGAAGCCAG
550

1767
ACUUUGGCAUGGACUGUAU
441
1767
ACUUUGGCAUGGACUGUAU
441
1785
AUACAGUCCAUGCCAAAGU
551

1785
UUUAUUUUCAAAUGGGACU
442
1785
UUUAUUUUCAAAUGGGACU
442
1803
AGUCCCAUUUGAAAAUAAA
552

1803
UUUUUCGUUUUUGUUUUUC
443
1803
UUUUUCGUUUUUGUUUUUC
443
1821
GAAAAACAAAAACGAAAAA
553

1821
CUGGGCAAGUUUUAUUGUG
444
1821
CUGGGCAAGUUUUAUUGUG
444
1839
CACAAUAAAACUUGCCCAG
554

1839
GAGAUUUUCUAAUUAUGAA
445
1839
GAGAUUUUCUAAUUAUGAA
445
1857
UUCAUAAUUAGAAAAUCUC
555

1857
AGCAAAAUUUCUUUUCUCC
446
1857
AGCAAAAUUUCUUUUCUCC
446
1875
GGAGAAAAGAAAUUUUGCU
556

1875
CACCAUGCUUUAUGUGAUA
447
1875
CACCAUGCUUUAUGUGAUA
447
1893
UAUCACAUAAAGCAUGGUG
557

1893
AGUAUUUAAAAUUGAUGUG
448
1893
AGUAUUUAAAAUUGAUGUG
448
1911
CACAUCAAUUUUAAAUACU
558

1911
GAGUUAUUAUGUCAAAAAA
449
1911
GAGUUAUUAUGUCAAAAAA
449
1929
UUUUUUGACAUAAUAACUC
559

1929
AACUGAUCUAUUAAAGAAG
450
1929
AACUGAUCUAUUAAAGAAG
450
1947
CUUCUUUAAUAGAUCAGUU
560

1947
GUAAUUGGCCUUUCUGAGC
451
1947
GUAAUUGGCCUUUCUGAGC
451
1965
GCUCAGAAAGGCCAAUUAC
561

1965
CUGAAAAAAAAAAAAAAAA
452
1965
CUGAAAAAAAAAAAAAAAA
452
1983
UUUUUUUUUUUUUUUUCAG
562

HDAC3: NM_003883.2

3
GGCGGCCGCGGGCGGCGGG
675
3
GGCGGCCGCGGGCGGCGGG
675
21
CCCGCCGCCCGCGGCCGCC
783

21
GCGGCGGAGGUGCGGGGCC
676
21
GCGGCGGAGGUGCGGGGCC
676
39
GGCCCCGCACCUCCGCCGC
784

39
CUGCUCCCGCCGGCACCAU
677
39
CUGCUCCCGCCGGCACCAU
677
57
AUGGUGCCGGCGGGAGCAG
785

57
UGGCCAAGACCGUGGCCUA
678
57
UGGCCAAGACCGUGGCCUA
678
75
UAGGCCACGGUCUUGGCCA
786

75
AUUUCUACGACCCCGACGU
679
75
AUUUCUACGACCCCGACGU
679
93
ACGUCGGGGUCGUAGAAAU
787

93
UGGGCAACUUCCACUACGG
680
93
UGGGCAACUUCCACUACGG
680
111
CCGUAGUGGAAGUUGCCCA
788

111
GAGCUGGACACCCUAUGAA
681
111
GAGCUGGACACCCUAUGAA
681
129
UUCAUAGGGUGUCCAGCUC
789

129
AGCCCCAUCGCCUGGCAUU
682
129
AGCCCCAUCGCCUGGCAUU
682
147
AAUGCCAGGCGAUGGGGCU
790

147
UGACCCAUAGCCUGGUCCU
683
147
UGACCCAUAGCCUGGUCCU
683
165
AGGACCAGGCUAUGGGUCA
791

165
UGCAUUACGGUCUCUAUAA
684
165
UGCAUUACGGUCUCUAUAA
684
183
UUAUAGAGACCGUAAUGCA
792

183
AGAAGAUGAUCGUCUUCAA
685
183
AGAAGAUGAUCGUCUUCAA
685
201
UUGAAGACGAUCAUCUUCU
793

201
AGCCAUACCAGGCCUCCCA
686
201
AGCCAUACCAGGCCUCCCA
686
219
UGGGAGGCCUGGUAUGGCU
794

219
AGCAUGACAUGUGCCGCUU
687
219
AGCAUGACAUGUGCCGCUU
687
237
AAGCGGCACAUGUCAUGCU
795

237
UCCACUCCGAGGACUACAU
688
237
UCCACUCCGAGGACUACAU
688
255
AUGUAGUCCUCGGAGUGGA
796

255
UUGACUUCCUGCAGAGAGU
689
255
UUGACUUCCUGCAGAGAGU
689
273
ACUCUCUGCAGGAAGUCAA
797

273
UCAGCCCCACCAAUAUGCA
690
273
UCAGCCCCACCAAUAUGCA
690
291
UGCAUAUUGGUGGGGCUGA
798

291
AAGGCUUCACCAAGAGUCU
691
291
AAGGCUUCACCAAGAGUCU
691
309
AGACUCUUGGUGAAGCCUU
799

309
UUAAUGCCUUCAACGUAGG
692
309
UUAAUGCCUUCAACGUAGG
692
327
CCUACGUUGAAGGCAUUAA
800

327
GCGAUGACUGCCCAGUGUU
693
327
GCGAUGACUGCCCAGUGUU
693
345
AACACUGGGCAGUCAUCGC
801

345
UUCCCGGGCUCUUUGAGUU
694
345
UUCCCGGGCUCUUUGAGUU
694
363
AACUCAAAGAGCCCGGGAA
802

363
UCUGCUCGCGUUACACAGG
695
363
UCUGCUCGCGUUACACAGG
695
381
CCUGUGUAACGCGAGCAGA
803

381
GCGCAUCUCUGCAAGGAGC
696
381
GCGCAUCUCUGCAAGGAGC
696
399
GCUCCUUGCAGAGAUGCGC
804

399
CAACCCAGCUGAACAACAA
697
399
CAACCCAGCUGAACAACAA
697
417
UUGUUGUUCAGCUGGGUUG
805

417
AGAUCUGUGAUAUUGCCAU
698
417
AGAUCUGUGAUAUUGCCAU
698
435
AUGGCAAUAUCACAGAUCU
806

435
UUAACUGGGCUGGUGGUCU
699
435
UUAACUGGGCUGGUGGUCU
699
453
AGACCACCAGCCCAGUUAA
807

453
UGCACCAUGCCAAGAAGUU
700
453
UGCACCAUGCCAAGAAGUU
700
471
AACUUCUUGGCAUGGUGCA
808

471
UUGAGGCCUCUGGCUUCUG
701
471
UUGAGGCCUCUGGCUUCUG
701
489
CAGAAGCCAGAGGCCUCAA
809

489
GCUAUGUCAACGACAUUGU
702
489
GCUAUGUCAACGACAUUGU
702
507
ACAAUGUCGUUGACAUAGC
810

507
UGAUUGGCAUCCUGGAGCU
703
507
UGAUUGGCAUCCUGGAGCU
703
525
AGCUCCAGGAUGCCAAUCA
811

525
UGCUCAAGUACCACCCUCG
704
525
UGCUCAAGUACCACCCUCG
704
543
CGAGGGUGGUACUUGAGCA
812

543
GGGUGCUCUACAUUGACAU
705
543
GGGUGCUCUACAUUGACAU
705
561
AUGUCAAUGUAGAGCACCC
813

561
UUGACAUCGACCAUGGUGA
706
561
UUGACAUCCACCAUGGUGA
706
579
UCACCAUGGUGGAUGUCAA
814

579
ACGGGGUUCAAGAAGCUUU
707
579
ACGGGGUUCAAGAAGCUUU
707
597
AAAGCUUCUUGAACCCCGU
815

597
UCUACCUCACUGACCGGGU
708
597
UCUACCUCACUGACCGGGU
708
615
ACCCGGUCAGUGAGGUAGA
816

615
UCAUGACGGUGUCCUUCCA
709
615
UCAUGACGGUGUCCUUCCA
709
633
UGGAAGGACACCGUCAUGA
817

633
ACAAAUACGGAAAUUACUU
710
633
ACAAAUACGGAAAUUACUU
710
651
AAGUAAUUUCCGUAUUUGU
818

651
UCUUCCCUGGCACAGGUGA
711
651
UCUUCCCUGGCACAGGUGA
711
669
UCACCUGUGCCAGGGAAGA
819

669
ACAUGUAUGAAGUCGGGGC
712
669
ACAUGUAUGAAGUCGGGGC
712
687
GCCCCGACUUCAUACAUGU
820

687
CAGAGAGUGGCCGCUACUA
713
687
CAGAGAGUGGCCGCUACUA
713
705
UAGUAGCGGCCACUCUCUG
821

705
ACUGUCUGAACGUGCCCCU
714
705
ACUGUCUGAACGUGCCCCU
714
723
AGGGGCACGUUCAGACAGU
822

723
UGCGGGAUGGCAUUGAUGA
715
723
UGCGGGAUGGCAUUGAUGA
715
741
UCAUCAAUGCCAUCCCGCA
823

741
ACCAGAGUUACAAGCACCU
716
741
ACCAGAGUUACAAGCACCU
716
759
AGGUGCUUGUAACUCUGGU
824

759
UUUUCCAGCCGGUUAUCAA
717
759
UUUUCCAGCCGGUUAUCAA
717
777
UUGAUAACCGGCUGGAAAA
825

777
ACCAGGUAGUGGACUUCUA
718
777
ACCAGGUAGUGGACUUCUA
718
795
UAGAAGUCCACUACCUGGU
826

795
ACCAACCCACGUGCAUUGU
719
795
ACCAACCCACGUGCAUUGU
719
813
ACAAUGCACGUGGGUUGGU
827

813
UGCUCCAGUGUGGAGCUGA
720
813
UGCUCCAGUGUGGAGCUGA
720
831
UCAGCUCCACACUGGAGCA
828

831
ACUCUCUGGGCUGUGAUCG
721
831
ACUCUCUGGGCUGUGAUCG
721
849
CGAUCACAGCCCAGAGAGU
829

849
GAUUGGGCUGCUUUAACCU
722
849
GAUUGGGCUGCUUUAACCU
722
867
AGGUUAAAGCAGCCCAAUC
830

867
UCAGCAUCCGAGGGCAUGG
723
867
UCAGCAUCCGAGGGCAUGG
723
885
CCAUGCCCUCGGAUGCUGA
831

885
GGGAAUGCGUUGAAUAUGU
724
885
GGGAAUGCGUUGAAUAUGU
724
903
ACAUAUUCAACGCAUUCCC
832

903
UCAAGAGCUUCAAUAUCCC
725
903
UCAAGAGCUUCAAUAUCCC
725
921
GGGAUAUUGAAGCUCUUGA
833

921
CUCUACUCGUGCUGGGUGG
726
921
CUCUACUCGUGCUGGGUGG
726
939
CCACCCAGCACGAGUAGAG
834

939
GUGGUGGUUAUACUGUCCG
727
939
GUGGUGGUUAUACUGUCCG
727
957
CGGACAGUAUAACCACCAC
835

957
GAAAUGUUGCCCGCUGCUG
728
957
GAAAUGUUGCCCGCUGCUG
728
975
CAGCAGCGGGCAACAUUUC
836

975
GGACAUAUGAGACAUCGCU
729
975
GGACAUAUGAGACAUCGCU
729
993
AGCGAUGUCUCAUAUGUCC
837

993
UGCUGGUAGAAGAGGCCAU
730
993
UGCUGGUAGAAGAGGCCAU
730
1011
AUGGCCUCUUCUACCAGCA
838

1011
UUAGUGAGGAGCUUCCCUA
731
1011
UUAGUGAGGAGCUUCCCUA
731
1029
UAGGGAAGCUCCUCACUAA
839

1029
AUAGUGAAUACUUCGAGUA
732
1029
AUAGUGAAUACUUCGAGUA
732
1047
UACUCGAAGUAUUCACUAU
840

1047
ACUUUGCCCCAGACUUCAC
733
1047
ACUUUGCCCCAGACUUCAC
733
1065
GUGAAGUCUGGGGCAAAGU
841

1065
CACUUCAUCCAGAUGUCAG
734
1065
CACUUCAUCCAGAUGUCAG
734
1083
CUGACAUCUGGAUGAAGUG
842

1083
GCACCCGCAUCGAGAAUCA
735
1083
GCACCCGCAUCGAGAAUCA
735
1101
UGAUUCUCGAUGCGGGUGC
843

1101
AGAACUCACGCCAGUAUCU
736
1101
AGAACUCACGCCAGUAUCU
736
1119
AGAUACUGGCGUGAGUUCU
844

1119
UGGACCAGAUCCGCCAGAC
737
1119
UGGACCAGAUCCGCCAGAC
737
1137
GUCUGGCGGAUCUGGUCCA
845

1137
CAAUCUUUGAAAACCUGAA
738
1137
CAAUCUUUGAAAACCUGAA
738
1155
UUCAGGUUUUCAAAGAUUG
846

1155
AGAUGCUGAACCAUGCACC
739
1155
AGAUGCUGAACCAUGCACC
739
1173
GGUGCAUGGUUCAGCAUCU
847

1173
CUAGUGUCCAGAUUCAUGA
740
1173
CUAGUGUCCAGAUUCAUGA
740
1191
UCAUGAAUCUGGACACUAG
848

1191
ACGUGCCUGCAGACCUCCU
741
1191
ACGUGCCUGCAGACCUCCU
741
1209
AGGAGGUCUGCAGGCACGU
849

1209
UGACCUAUGACAGGACUGA
742
1209
UGACCUAUGACAGGACUGA
742
1227
UCAGUCCUGUCAUAGGUCA
850

1227
AUGAGGCUGAUGCAGAGGA
743
1227
AUGAGGCUGAUGCAGAGGA
743
1245
UCCUCUGCAUCAGCCUCAU
851

1245
AGAGGGGUCCUGAGGAGAA
744
1245
AGAGGGGUCCUGAGGAGAA
744
1263
UUCUCCUCAGGACCCCUCU
852

1263
ACUAUAGCAGGCCAGAGGC
745
1263
ACUAUAGCAGGCCAGAGGC
745
1281
GCCUCUGGCCUGCUAUAGU
853

1281
CACCCAAUGAGUUCUAUGA
746
1281
CACCCAAUGAGUUCUAUGA
746
1299
UCAUAGAACUCAUUGGGUG
854

1299
AUGGAGACCAUGACAAUGA
747
1299
AUGGAGACCAUGACAAUGA
747
1317
UCAUUGUCAUGGUCUCCAU
855

1317
ACAAGGAAAGCGAUGUGGA
748
1317
ACAAGGAAAGCGAUGUGGA
748
1335
UCCACAUCGCUUUCCUUGU
856

1335
AGAUUUAAGAGUGGCUUGG
749
1335
AGAUUUAAGAGUGGCUUGG
749
1353
CCAAGCCACUCUUAAAUCU
857

1353
GGAUGCUGUGUCCCAAGGA
750
1353
GGAUGCUGUGUCCCAAGGA
750
1371
UCCUUGGGACACAGCAUCC
858

1371
AAUUUCUUUUCACCUCUUG
751
1371
AAUUUCUUUUCACCUCUUG
751
1389
CAAGAGGUGAAAAGAAAUU
859

1389
GGUUGGGCUGGAGGGAAAA
752
1389
GGUUGGGCUGGAGGGAAAA
752
1407
UUUUCCCUCCAGCCCAACC
860

1407
AGGAGUGGCUCCUAGAGUC
753
1407
AGGAGUGGCUCCUAGAGUC
753
1425
GACUCUAGGAGCCACUCCU
861

1425
CCUGGGGGUCACCCCAGGG
754
1425
CCUGGGGGUCACCCCAGGG
754
1443
CCCUGGGGUGACCCCCAGG
862

1443
GCUUUUGCUGACUCUGGGA
755
1443
GCUUUUGCUGACUCUGGGA
755
1461
UCCCAGAGUCAGCAAAAGC
863

1461
AAAGAGUCUGGAGACCACA
756
1461
AAAGAGUCUGGAGACCACA
756
1479
UGUGGUCUCCAGACUCUUU
864

1479
AUUUGGUUCUCGAACCAUC
757
1479
AUUUGGUUCUCGAACCAUC
757
1497
GAUGGUUCGAGAACCAAAU
865

1497
CUACCUGCUUUUCCUCUCU
758
1497
CUACCUGCUUUUCCUCUCU
758
1515
AGAGAGGAAAAGCAGGUAG
866

1515
UCUCCCAAGGCCUGACAAU
759
1515
UCUCCCAAGGCCUGACAAU
759
1533
AUUGUCAGGCCUUGGGAGA
867

1533
UGGUACCUAUUAGGGAUGG
760
1533
UGGUACCUAUUAGGGAUGG
760
1551
CCAUCCCUAAUAGGUACCA
868

1551
GAGAUACAGACAAGGAUAG
761
1551
GAGAUACAGACAAGGAUAG
761
1569
CUAUCCUUGUCUGUAUCUC
869

1569
GCUAUCUGGGACAUUAUUG
762
1569
GCUAUCUGGGACAUUAUUG
762
1587
CAAUAAUGUCCCAGAUAGC
870

1587
GGCAGUGGGCCCUGGAGGC
763
1587
GGCAGUGGGCCCUGGAGGC
763
1605
GCCUCCAGGGCCCACUGCC
871

1605
CCAGUCCCUAGCCCCCCUU
764
1605
CCAGUCCCUAGCCCCCCUU
764
1623
AAGGGGGGCUAGGGACUGG
872

1623
UGCCCCUUAUUUCUUCCCU
765
1623
UGCCCCUUAUUUCUUCCCU
765
1641
AGGGAAGAAAUAAGGGGCA
873

1641
UGCUUCCCUCGAACCCAGA
766
1641
UGCUUCCCUCGAACCCAGA
766
1659
UCUGGGUUCGAGGGAAGCA
874

1659
AGAUUUUUGAGGGAUGAAC
767
1659
AGAUUUUUGAGGGAUGAAC
767
1677
GUUCAUCCCUCAAAAAUCU
875

1677
CGGGUAGACAAGGACUGAG
768
1677
CGGGUAGACAAGGACUGAG
768
1695
CUCAGUCCUUGUCUACCCG
876

1695
GAUUGCCUCUGACUUCCUC
769
1695
GAUUGCCUCUGACUUCCUC
769
1713
GAGGAAGUCAGAGGCAAUC
877

1713
CCUCCCCUGGGUUCUGACU
770
1713
CCUCCCCUGGGUUCUGACU
770
1731
AGUCAGAACCCAGGGGAGG
878

1731
UUCUUCCUCCCCUUGCUUC
771
1731
UUCUUCCUCCCCUUGCUUC
771
1749
GAAGCAAGGGGAGGAAGAA
879

1749
CCAGGGAAGAUGAAGAGAG
772
1749
CCAGGGAAGAUGAAGAGAG
772
1767
CUCUCUUCAUCUUCCCUGG
880

1767
GAGAGAUUUGGAAGGGGCU
773
1767
GAGAGAUUUGGAAGGGGCU
773
1785
AGCCCCUUCCAAAUCUCUC
881

1785
UCUGGCUCCCUAACACCUG
774
1785
UCUGGCUCCCUAACACCUG
774
1803
CAGGUGUUAGGGAGCCAGA
882

1803
GAAUCCCAGAUGAUGGGAA
775
1803
GAAUCCCAGAUGAUGGGAA
775
1821
UUCCCAUCAUCUGGGAUUC
883

1821
AGUAUGUUUUCAAGUGUGG
776
1821
AGUAUGUUUUCAAGUGUGG
776
1839
CCACACUUGAAAACAUACU
884

1839
GGGAGGAUAUGAAAAUGUU
777
1839
GGGAGGAUAUGAAAAUGUU
777
1857
AACAUUUUCAUAUCCUCCC
885

1857
UCUGUUCUCACUUUUGGCU
778
1857
UCUGUUCUCACUUUUGGCU
778
1875
AGCCAAAAGUGAGAACAGA
886

1875
UUUAUGUCCAUUUUACCAC
779
1875
UUUAUGUCCAUUUUACCAC
779
1893
GUGGUAAAAUGGACAUAAA
887

1893
CUGUUUUUAUCCAAUAAAC
780
1893
CUGUUUUUAUCCAAUAAAC
780
1911
GUUUAUUGGAUAAAAACAG
888

1911
CUAAGUCGGUAUUUUUUGU
781
1911
CUAAGUCGGUAUUUUUUGU
781
1929
ACAAAAAAUACCGACUUAG
889

1929
UACCUUUAAAAAAAAAAAA
782
1929
UACCUUUAAAAAAAAAAAA
782
1947
UUUUUUUUUUUUAAAGGUA
890

HDAC4: NM_006037.2

3
AGGUUGUGGGGCCGCCGCC
1003
3
AGGUUGUGGGGCCGCCGCC
1003
21
GGCGGCGGCCCCACAACCU
1472

21
CGCGGAGCACCGUCCCCGC
1004
21
CGCGGAGCACCGUCCCCGC
1004
39
GCGGGGACGGUGCUCCGCG
1473

39
CCGCCGCCCGAGCCCGAGC
1005
39
CCGCCGCCCGAGCCCGAGC
1005
57
GCUCGGGCUCGGGCGGCGG
1474

57
CCCGAGCCCGCGCACCCGC
1006
57
CCCGAGCCCGCGCACCCGC
1006
75
GCGGGUGCGCGGGCUCGGG
1475

75
CCCGCGCCGCCGCCGCCGC
1007
75
CCCGCGCCGCCGCCGCCGC
1007
93
GCGGCGGCGGCGGCGCGGG
1476

93
CCGCCCGAACAGCCUCCCA
1008
93
CCGCCCGAACAGCCUCCCA
1008
111
UGGGAGGCUGUUCGGGCGG
1477

111
AGCCUGGGCCCCCGGCGGC
1009
111
AGCCUGGGCCCCCGGCGGC
1009
129
GCCGCCGGGGGCCCAGGCU
1478

129
CGCCGUGGCCGCGUCCCGG
1010
129
CGCCGUGGCCGCGUCCCGG
1010
147
CCGGGACGCGGCCACGGCG
1479

147
GCUGUCGCCGCCCGAGCCC
1011
147
GCUGUCGCCGCCCGAGCCC
1011
165
GGGCUCGGGCGGCGACAGC
1480

165
CGAGCCCGCGCGCCGGCGG
1012
165
CGAGCCCGCGCGCCGGCGG
1012
183
CCGCCGGCGCGCGGGCUCG
1481

183
GGUGGCGGCGCAGGCUGAG
1013
183
GGUGGCGGCGCAGGCUGAG
1013
201
CUCAGCCUGCGCCGCCACC
1482

201
GGAGAUGCGGCGCGGAGCG
1014
201
GGAGAUGCGGCGCGGAGCG
1014
219
CGCUCCGCGCCGCAUCUCC
1483

219
GCCGGAGCAGGGCUAGAGC
1015
219
GCCGGAGCAGGGCUAGAGC
1015
237
GCUCUAGCCCUGCUCCGGC
1484

237
CCGGCCGCCGCCGCCCGCC
1016
237
CCGGCCGCCGCCGCCCGCC
1016
255
GGCGGGCGGCGGCGGCCGG
1485

255
CGCGGUAAGCGCAGCCCCG
1017
255
CGCGGUAAGCGCAGCCCCG
1017
273
CGGGGCUGCGCUUACCGCG
1486

273
GGCCCGGCGCCCGCGGGCC
1018
273
GGCCCGGCGCCCGCGGGCC
1018
291
GGCCCGCGGGCGCCGGGCC
1487

291
CAUUGUCCGCCGCCCGCCC
1019
291
CAUUGUCCGCCGCCCGCCC
1019
309
GGGCGGGCGGCGGACAAUG
1488

309
CCGCGCCCCGCGCAGCCUG
1020
309
CCGCGCCCCGCGCAGCCUG
1020
327
CAGGCUGCGCGGGGCGCGG
1489

327
GCAGGCCUUGGAGCCCGCG
1021
327
GCAGGCCUUGGAGCCCGCG
1021
345
CGCGGGCUCCAAGGCCUGC
1490

345
GGCAGGUGGACGCCGCCGG
1022
345
GGCAGGUGGACGCCGCCGG
1022
363
CCGGGGGCGUCCACCUGCC
1491

363
GUCCACACCCGCCCCGCGC
1023
363
GUCCACACCCGCCCCGCGC
1023
381
GCGCGGGGCGGGUGUGGAC
1492

381
CGCGGCCGUGGGAGGCGGG
1024
381
CGCGGCCGUGGGAGGCGGG
1024
399
CCCGCCUCCCACGGCCGCG
1493

399
GGGCCAGCGCUGGCCGCGC
1025
399
GGGCCAGCGCUGGCCGCGC
1025
417
GCGCGGCCAGCGCUGGCCC
1494

417
CGCCGUGGGACCCGCCGGU
1026
417
CGCCGUGGGACCCGCCGGU
1026
435
ACCGGCGGGUCCCACGGCG
1495

435
UCCCCAGGGCCGCCCGGCC
1027
435
UCCCCAGGGCCGCCCGGCC
1027
453
GGCCGGGCGGCCCUGGGGA
1496

453
CCCUUCUGGACCUUUCCAC
1028
453
CCCUUCUGGACCUUUCCAC
1028
471
GUGGAAAGGUCCAGAAGGG
1497

471
CCCGCGCCGCGAGGCGGCU
1029
471
CCCGCGCCGCGAGGCGGCU
1029
489
AGCCGCCUCGCGGCGCGGG
1498

489
UUCGCCCGCCGGGGCGGGG
1030
489
UUCGCCCGCCGGGGCGGGG
1030
507
CCCCGCCCCGGCGGGCGAA
1499

507
GGCGCGGGGGUGGGCACGG
1031
507
GGCGCGGGGGUGGGCACGG
1031
525
CCGUGCCCACCCCCGCGCC
1500

525
GCAGGCAGCGGCGCCGUCU
1032
525
GCAGGCAGCGGCGCCGUCU
1032
543
AGACGGCGCCGCUGCCUGC
1501

543
UCCCGGUGCGGGGCCCGCG
1033
543
UCCCGGUGCGGGGCCCGCG
1033
561
CGCGGGCCCCGCACCGGGA
1502

561
GCCCCCCGAGCAGGUUCAU
1034
561
GCCCCCCGAGCAGGUUCAU
1034
579
AUGAACCUGCUCGGGGGGC
1503

579
UCUGCAGAAGCCAGCGGAC
1035
579
UCUGCAGAAGCCAGCGGAC
1035
597
GUCCGCUGGCUUCUGCAGA
1504

597
CGCCUCUGUUCAACUUGUG
1036
597
CGCCUCUGUUCAACUUGUG
1036
615
CACAAGUUGAACAGAGGCG
1505

615
GGGUUACCUGGCUCAUGAG
1037
615
GGGUUACCUGGCUCAUGAG
1037
633
CUCAUGAGCCAGGUAACCC
1506

633
GACCUUGCCGGCGAGGCUC
1038
633
GACCUUGCCGGCGAGGCUC
1038
651
GAGCCUCGCCGGCAAGGUC
1507

651
CGGCGCUUGAACGUCUGUG
1039
651
CGGCGCUUGAACGUCUGUG
1039
669
CACAGACGUUCAAGCGCCG
1508

669
GACCCAGCCCUCACCGUCC
1040
669
GACCCAGCCCUCACCGUCC
1040
687
GGACGGUGAGGGCUGGGUC
1509

687
CCGGUACUUGUAUGUGUUG
1041
687
CCGGUACUUGUAUGUGUUG
1041
705
CAACACAUACAAGUACCGG
1510

705
GGUGGGAGUUUGGAGCUCG
1042
705
GGUGGGAGUUUGGAGCUCG
1042
723
CGAGCUCCAAACUCCCACC
1511

723
GUUGGAGCUAUCGUUUCCG
1043
723
GUUGGAGCUAUCGUUUCCG
1043
741
CGGAAACGAUAGCUCCAAC
1512

741
GUGGAAAUUUUGAGCCAUU
1044
741
GUGGAAAUUUUGAGCCAUU
1044
759
AAUGGCUCAAAAUUUCCAC
1513

759
UUCGAAUCACUUAAAGGAG
1045
759
UUCGAAUCACUUAAAGGAG
1045
777
CUCCUUUAAGUGAUUCGAA
1514

777
GUGGACAUUGCUAGCAAUG
1046
777
GUGGACAUUGCUAGCAAUG
1046
795
CAUUGCUAGCAAUGUCCAC
1515

795
GAGCUCCCAAAGCCAUCCA
1047
795
GAGCUCCCAAAGCCAUCCA
1047
813
UGGAUGGCUUUGGGAGCUC
1516

813
AGAUGGACUUUCUGGCCGA
1048
813
AGAUGGACUUUCUGGCCGA
1048
831
UCGGCCAGAAAGUCCAUCU
1517

831
AGACCAGCCAGUGGAGCUG
1049
831
AGACCAGCCAGUGGAGCUG
1049
849
CAGCUCCACUGGCUGGUCU
1518

849
GCUGAAUCCUGCCCGCGUG
1050
849
GCUGAAUCCUGCCCGCGUG
1050
867
CACGCGGGCAGGAUUCAGC
1519

867
GAACCACAUGCCCAGCACG
1051
867
GAACCACAUGCCCAGCACG
1051
885
CGUGCUGGGCAUGUGGUUC
1520

885
GGUGGAUGUGGCCACGGCG
1052
885
GGUGGAUGUGGCCACGGCG
1052
903
CGCCGUGGCCACAUCCACC
1521

903
GCUGCCUCUGCAAGUGGCC
1053
903
GCUGCCUCUGCAAGUGGCC
1053
921
GGCCACUUGCAGAGGCAGC
1522

921
CCCCUCGGCAGUGCCCAUG
1054
921
CCCCUCGGCAGUGCCCAUG
1054
939
CAUGGGCACUGCCGAGGGG
1523

939
GGACCUGCGCCUGGACCAC
1055
939
GGACCUGCGCCUGGACCAC
1055
957
GUGGUCCAGGCGCAGGUCC
1524

957
CCAGUUCUCACUGCCUGUG
1056
957
CCAGUUCUCACUGCCUGUG
1056
975
CACAGGCAGUGAGAACUGG
1525

975
GGCAGAGCCGGCCCUGCGG
1057
975
GGCAGAGCCGGCCCUGCGG
1057
993
CCGCAGGGCCGGCUCUGCC
1526

993
GGAGCAGCAGCUGCAGCAG
1058
993
GGAGCAGCAGCUGCAGCAG
1058
1011
CUGCUGCAGCUGCUGCUCC
1527

1011
GGAGCUCCUGGCGCUCAAG
1059
1011
GGAGCUCCUGGCGCUCAAG
1059
1029
CUUGAGCGCCAGGAGCUCC
1528

1029
GCAGAAGCAGCAGAUCCAG
1060
1029
GCAGAAGCAGCAGAUCCAG
1060
1047
CUGGAUCUGCUGCUUCUGC
1529

1047
GAGGCAGAUCCUCAUCGCU
1061
1047
GAGGCAGAUCCUCAUCGCU
1061
1065
AGCGAUGAGGAUCUGCCUC
1530

1065
UGAGUUCCAGAGGCAGCAC
1062
1065
UGAGUUCCAGAGGCAGCAC
1062
1083
GUGCUGCCUCUGGAACUCA
1531

1083
CGAGCAGCUCUCCCGGCAG
1063
1083
CGAGCAGCUCUCCCGGCAG
1063
1101
CUGCCGGGAGAGCUGCUCG
1532

1101
GCACGAGGCGCAGCUCCAC
1064
1101
GCACGAGGCGCAGCUCCAC
1064
1119
GUGGAGCUGCGCCUCGUGC
1533

1119
CGAGCACAUCAAGCAACAA
1065
1119
CGAGCACAUCAAGCAACAA
1065
1137
UUGUUGCUUGAUGUGCUCG
1534

1137
ACAGGAGAUGCUGGCCAUG
1066
1137
ACAGGAGAUGCUGGCCAUG
1066
1155
CAUGGCCAGCAUCUCCUGU
1535

1155
GAAGCACCAGCAGGAGCUG
1067
1155
GAAGCACCAGCAGGAGCUG
1067
1173
CAGCUCCUGCUGGUGCUUC
1536

1173
GCUGGAACACCAGCGGAAG
1068
1173
GCUGGAACACCAGCGGAAG
1068
1191
CUUCCGCUGGUGUUCCAGC
1537

1191
GCUGGAGAGGCACCGCCAG
1069
1191
GCUGGAGAGGCACCGCCAG
1069
1209
CUGGCGGUGCCUCUCCAGC
1538

1209
GGAGCAGGAGCUGGAGAAG
1070
1209
GGAGCAGGAGCUGGAGAAG
1070
1227
CUUCUCCAGCUCCUGCUCC
1539

1227
GCAGCACCGGGAGCAGAAG
1071
1227
GCAGCACCGGGAGCAGAAG
1071
1245
CUUCUGCUCCCGGUGCUGC
1540

1245
GCUGCAGCAGCUCAAGAAC
1072
1245
GCUGCAGCAGCUCAAGAAC
1072
1263
GUUCUUGAGCUGCUGCAGC
1541

1263
CAAGGAGAAGGGCAAAGAG
1073
1263
CAAGGAGAAGGGCAAAGAG
1073
1281
CUCUUUGCCCUUCUCCUUG
1542

1281
GAGUGCCGUGGCCAGCACA
1074
1281
GAGUGCCGUGGCCAGCACA
1074
1299
UGUGCUGGCCACGGCACUC
1543

1299
AGAAGUGAAGAUGAAGUUA
1075
1299
AGAAGUGAAGAUGAAGUUA
1075
1317
UAACUUCAUCUUCACUUCU
1544

1317
ACAAGAAUUUGUCCUCAAU
1076
1317
ACAAGAAUUUGUCCUCAAU
1076
1335
AUUGAGGACAAAUUCUUGU
1545

1335
UAAAAAGAAGGCGCUGGCC
1077
1335
UAAAAAGAAGGCGCUGGCC
1077
1353
GGCCAGCGCCUUCUUUUUA
1546

1353
CCACCGGAAUCUGAACCAC
1078
1353
CCACCGGAAUCUGAACCAC
1078
1371
GUGGUUCAGAUUCCGGUGG
1547

1371
CUGCAUUUCCAGCGACCCU
1079
1371
CUGCAUUUCCAGCGAGCCU
1079
1389
AGGGUCGCUGGAAAUGCAG
1548

1389
UCGCUACUGGUACGGGAAA
1080
1389
UCGCUACUGGUACGGGAAA
1080
1407
UUUCCCGUACCAGUAGCGA
1549

1407
AACGCAGCACAGUUCCCUU
1081
1407
AACGCAGCACAGUUCCCUU
1081
1425
AAGGGAACUGUGCUGCGUU
1550

1425
UGACCAGAGUUCUCCACCC
1082
1425
UGACCAGAGUUCUCCACCC
1082
1443
GGGUGGAGAACUCUGGUCA
1551

1443
CCAGAGCGGAGUGUCGACC
1083
1443
CCAGAGCGGAGUGUCGACC
1083
1461
GGUCGACACUCCGCUCUGG
1552

1461
CUCCUAUAACCACCCGGUC
1084
1461
CUCCUAUAACCACCCGGUC
1084
1479
GACCGGGUGGUUAUAGGAG
1553

1479
CCUGGGAAUGUACGACGCC
1085
1479
CCUGGGAAUGUACGACGCC
1085
1497
GGCGUCGUACAUUCCCAGG
1554

1497
CAAAGAUGACUUCCCUCUU
1086
1497
CAAAGAUGACUUCCCUCUU
1086
1515
AAGAGGGAAGUCAUCUUUG
1555

1515
UAGGAAAACAGCUUCUGAA
1087
1515
UAGGAAAACAGCUUCUGAA
1087
1533
UUCAGAAGCUGUUUUCCUA
1556

1533
ACCGAAUCUGAAAUUAGGG
1088
1533
ACCGAAUCUGAAAUUACGG
1088
1551
CCGUAAUUUCAGAUUCGGU
1557

1551
GUCCAGGCUAAAGCAGAAA
1089
1551
GUCCAGGCUAAAGCAGAAA
1089
1569
UUUCUGCUUUAGCCUGGAC
1558

1569
AGUGGCCGAAAGACGGAGC
1090
1569
AGUGGCCGAAAGACGGAGC
1090
1587
GCUCCGUCUUUCGGCCACU
1559

1587
CAGCCCCCUGUUACGCAGG
1091
1587
CAGCCCCCUGUUACGCAGG
1091
1605
CCUGCGUAACAGGGGGCUG
1560

1605
GAAAGACGGGCCAGUGGUC
1092
1605
GAAAGACGGGCCAGUGGUC
1092
1623
GACCACUGGCCCGUCUUUC
1561

1623
CACUGCUCUAAAAAAGCGU
1093
1623
CACUGCUCUAAAAAAGCGU
1093
1641
ACGCUUUUUUAGAGCAGUG
1562

1641
UCCGUUGGAUGUCACAGAC
1094
1641
UCCGUUGGAUGUCACAGAC
1094
1659
GUCUGUGACAUCCAACGGA
1563

1659
CUCCGCGUGCAGCAGCGCC
1095
1659
CUCCGCGUGCAGCAGCGCC
1095
1677
GGCGCUGCUGCACGCGGAG
1564

1677
CCCAGGCUCCGGACCCAGC
1096
1677
CCCAGGCUCCGGACCCAGC
1096
1695
GCUGGGUCCGGAGCCUGGG
1565

1695
CUCACCCAACAACAGCUCC
1097
1695
CUCACCCAACAACAGCUCC
1097
1713
GGAGCUGUUGUUGGGUGAG
1566

1713
CGGGAGCGUCAGCGCGGAG
1098
1713
CGGGAGCGUCAGCGCGGAG
1098
1731
CUCCGCGCUGACGCUCCCG
1567

1731
GAACGGUAUCGCGCCCGCC
1099
1731
GAACGGUAUCGCGCCCGCC
1099
1749
GGCGGGCGCGAUACCGUUC
1568

1749
CGUCCCCAGCAUCCCGGCG
1100
1749
CGUCCCCAGCAUCCCGGCG
1100
1767
CGCCGGGAUGCUGGGGACG
1569

1767
GGAGACGAGUUUGGCGCAC
1101
1767
GGAGACGAGUUUGGCGCAC
1101
1785
GUGCGCCAAACUCGUCUCC
1570

1785
CAGACUUGUGGCACGAGAA
1102
1785
CAGACUUGUGGCACGAGAA
1102
1803
UUCUCGUGCCACAAGUCUG
1571

1803
AGGCUCGGCCGCUCCACUU
1103
1803
AGGCUCGGCCGCUCCACUU
1103
1821
AAGUGGAGCGGCCGAGCCU
1572

1821
UCCCCUCUACACAUCGCCA
1104
1821
UCCCCUCUACACAUCGCCA
1104
1839
UGGCGAUGUGUAGAGGGGA
1573

1839
AUCCUUGCCCAACAUCACG
1105
1839
AUCCUUGCCCAACAUCACG
1105
1857
CGUGAUGUUGGGCAAGGAU
1574

1857
GCUGGGCCUGCCUGCCACC
1106
1857
GCUGGGCCUGCCUGCCACC
1106
1875
GGUGGCAGGCAGGCCCAGC
1575

1875
CGGCCCCUCUGCGGGCACG
1107
1875
CGGCCCCUCUGCGGGCACG
1107
1893
CGUGCCCGCAGAGGGGCCG
1576

1893
GGCGGGCCAGCAGGACACC
1108
1893
GGCGGGCCAGCAGGACACC
1108
1911
GGUGUCCUGCUGGCCCGCC
1577

1911
CGAGAGACUCACCCUUCCC
1109
1911
CGAGAGACUCACCCUUCCC
1109
1929
GGGAAGGGUGAGUCUCUCG
1578

1929
CGCCCUCCAGCAGAGGCUC
1110
1929
CGCCCUCCAGCAGAGGCUC
1110
1947
GAGCCUCUGCUGGAGGGCG
1579

1947
CUCCCUUUUCCCCGGCACC
1111
1947
CUCCCUUUUCCCCGGCACC
1111
1965
GGUGCCGGGGAAAAGGGAG
1580

1965
CCACCUCACUCCCUACCUG
1112
1965
CCACCUCACUCCCUACCUG
1112
1983
CAGGUAGGGAGUGAGGUGG
1581

1983
GAGCACCUCGCCCUUGGAG
1113
1983
GAGCACCUCGCCCUUGGAG
1113
2001
CUCCAAGGGCGAGGUGCUC
1582

2001
GCGGGACGGAGGGGCAGCG
1114
2001
GCGGGACGGAGGGGCAGCG
1114
2019
CGCUGCCCCUCCGUCCCGC
1583

2019
GCACAGCCCUCUUCUGCAG
1115
2019
GCACAGCCCUCUUCUGCAG
1115
2037
CUGCAGAAGAGGGCUGUGC
1584

2037
GCACAUGGUCUUACUGGAG
1116
2037
GCACAUGGUCUUACUGGAG
1116
2055
CUCCAGUAAGACCAUGUGC
1585

2055
GCAGCCACCGGCACAAGCA
1117
2055
GCAGCCACCGGCACAAGCA
1117
2073
UGCUUGUGCCGGUGGCUGC
1586

2073
ACCCCUCGUCACAGGCCUG
1118
2073
ACCCCUCGUCACAGGCCUG
1118
2091
CAGGCCUGUGACGAGGGGU
1587

2091
GGGAGCACUGCCCCUCCAC
1119
2091
GGGAGCACUGCCCCUCCAC
1119
2109
GUGGAGGGGCAGUGCUCCC
1588

2109
CGCACAGUCCUUGGUUGGU
1120
2109
CGCAGAGUCCUUGGUUGGU
1120
2127
ACCAACCAAGGACUGUGCG
1589

2127
UGCAGACCGGGUGUCCCCC
1121
2127
UGCAGACCGGGUGUCCCCC
1121
2145
GGGGGACACCCGGUCUGCA
1590

2145
CUCCAUCCAGAAGCUGCGG
1122
2145
CUCCAUCCACAAGCUGCGG
1122
2163
CCGCAGCUUGUGGAUGGAG
1591

2163
GCAGCACCGCCCACUGGGG
1123
2163
GCAGCACCGCCCACUGGGG
1123
2181
CCCCAGUGGGCGGUGCUGC
1592

2181
GCGGACCCAGUCGGCCCCG
1124
2181
GCGGACCCAGUCGGCCCCG
1124
2199
CGGGGCCGACUGGGUCCGC
1593

2199
GCUGCCCCAGAACGCCCAG
1125
2199
GCUGCCCCAGAACGCCCAG
1125
2217
CUGGGCGUUCUGGGGCAGC
1594

2217
GGCUCUGCAGCACCUGGUC
1126
2217
GGCUCUGCAGCACCUGGUC
1126
2235
GACCAGGUGCUGCAGAGCC
1595

2235
CAUCCAGCAGCAGCAUCAG
1127
2235
CAUCCAGCAGCAGCAUCAG
1127
2253
CUGAUGCUGCUGCUGGAUG
1596

2253
GCAGUUUCUGGAGAAACAC
1128
2253
GCAGUUUCUGGAGAAACAC
1128
2271
GUGUUUCUCCAGAAACUGC
1597

2271
CAAGCAGCAGUUCCAGCAG
1129
2271
CAAGCAGCAGUUCCAGCAG
1129
2289
CUGCUGGAACUGCUGCUUG
1598

2289
GCAGCAACUGCAGAUGAAC
1130
2289
GCAGCAACUGCAGAUGAAC
1130
2307
GUUCAUCUGCAGUUGCUGC
1599

2307
CAAGAUCAUCCCCAAGCCA
1131
2307
CAAGAUCAUCCCCAAGCCA
1131
2325
UGGCUUGGGGAUGAUCUUG
1600

2325
AAGCGAGCCAGCCCGGCAG
1132
2325
AAGCGAGCCAGCCCGGCAG
1132
2343
CUGCCGGGCUGGCUCGCUU
1601

2343
GCCGGAGAGCCACCCGGAG
1133
2343
GCCGGAGAGCCACCCGGAG
1133
2361
CUCCGGGUGGCUCUCCGGC
1602

2361
GGAGACGGAGGAGGAGCUC
1134
2361
GGAGACGGAGGAGGAGCUC
1134
2379
GAGCUCCUCCUCCGUCUCC
1603

2379
CCGUGAGCACCAGGCUCUG
1135
2379
CCGUGAGCACCAGGCUCUG
1135
2397
CAGAGCCUGGUGCUCACGG
1604

2397
GCUGGACGAGCCCUACCUG
1136
2397
GCUGGACGAGCCCUACCUG
1136
2415
CAGGUAGGGCUCGUCCAGC
1605

2415
GGACCGGCUGCCGGGGCAG
1137
2415
GGACCGGCUGCCGGGGCAG
1137
2433
CUGCCCCGGCAGCCGGUCC
1606

2433
GAAGGAGGCGCACGCACAG
1138
2433
GAAGGAGGCGCACGCACAG
1138
2451
CUGUGCGUGCGCCUCCUUC
1607

2451
GGCCGGCGUGCAGGUGAAG
1139
2451
GGCCGGCGUGCAGGUGAAG
1139
2469
CUUCACCUGCACGCCGGCC
1608

2469
GCAGGAGCCCAUUGAGAGC
1140
2469
GCAGGAGCCCAUUGAGAGC
1140
2487
GCUCUCAAUGGGCUCCUGC
1609

2487
CGAUGAGGAAGAGGCAGAG
1141
2487
CGAUGAGGAAGAGGCAGAG
1141
2505
CUCUGCCUCUUCCUCAUCG
1610

2505
GCCCCCACGGGAGGUGGAG
1142
2505
GCCCCCACGGGAGGUGGAG
1142
2523
CUCCACCUCCCGUGGGGGC
1611

2523
GCCGGGCCAGCGCCAGCCC
1143
2523
GCCGGGCCAGCGCCAGCCC
1143
2541
GGGCUGGCGCUGGCCCGGC
1612

2541
CAGUGAGCAGGAGCUGCUC
1144
2541
CAGUGAGCAGGAGCUGCUC
1144
2559
GAGCAGCUCCUGCUCACUG
1613

2559
CUUCAGACAGCAAGCCCUC
1145
2559
CUUCAGACAGCAAGCCCUC
1145
2577
GAGGGCUUGCUGUCUGAAG
1614

2577
CCUGCUGGAGCAGCAGCGG
1146
2577
CCUGCUGGAGCAGCAGCGG
1146
2595
CCGCUGCUGCUCCAGCAGG
1615

2595
GAUCCACCAGCUGAGGAAC
1147
2595
GAUCCACCAGCUGAGGAAC
1147
2613
GUUCCUCAGCUGGUGGAUC
1616

2613
CUACCAGGCGUCCAUGGAG
1148
2613
CUACCAGGCGUCCAUGGAG
1148
2631
CUCCAUGGACGCCUGGUAG
1617

2631
GGCCGCCGGCAUCCCCGUG
1149
2631
GGCCGCCGGCAUCCCCGUG
1149
2649
CACGGGGAUGCCGGCGGCC
1618

2649
GUCCUUCGGCGGCCACAGG
1150
2649
GUCCUUCGGCGGCCACAGG
1150
2667
CCUGUGGCCGCCGAAGGAC
1619

2667
GCCUCUGUCCCGGGCGCAG
1151
2667
GCCUCUGUCCCGGGCGCAG
1151
2685
CUGCGCCCGGGACAGAGGC
1620

2685
GUCCUCACCCGCGUCUGCC
1152
2685
GUCCUCACCCGCGUCUGCC
1152
2703
GGCAGACGCGGGUGAGGAC
1621

2703
CACCUUCCCCGUGUCUGUG
1153
2703
CACCUUCCCCGUGUCUGUG
1153
2721
CACAGACACGGGGAAGGUG
1622

2721
GCAGGAGCCCCCCACCAAG
1154
2721
GCAGGAGCCCCCCACCAAG
1154
2739
CUUGGUGGGGGGCUCCUGC
1623

2739
GCCGAGGUUCACGACAGGC
1155
2739
GCCGAGGUUCACGACAGGC
1155
2757
GGCUGUCGUGAACCUCGGC
1624

2757
CCUCGUGUAUGACACGCUG
1156
2757
CCUCGUGUAUGACACGCUG
1156
2775
CAGCGUGUCAUACACGAGG
1625

2775
GAUGCUGAAGCACCAGUGC
1157
2775
GAUGCUGAAGCACCAGUGC
1157
2793
GCACUGGUGCUUCAGCAUC
1626

2793
CACCUGCGGGAGUAGCAGC
1158
2793
CACCUGCGGGAGUAGCAGC
1158
2811
GCUGCUACUCCCGCAGGUG
1627

2811
CAGCCACCCCGAGCACGCC
1159
2811
CAGCCACCCCGAGCACGCC
1159
2829
GGCGUGCUCGGGGUGGCUG
1628

2829
CGGGAGGAUCCAGAGCAUC
1160
2829
CGGGAGGAUCCAGAGCAUC
1160
2847
GAUGCUCUGGAUCCUCCCG
1629

2847
CUGGUCCCGCCUGCAGGAG
1161
2847
CUGGUCCCGCCUGCAGGAG
1161
2865
CUCCUGCAGGCGGGACCAG
1630

2865
GACGGGCCUCCGGGGCAAA
1162
2865
GACGGGCCUCCGGGGCAAA
1162
2883
UUUGCCCCGGAGGCCCGUC
1631

2883
AUGCGAGUGCAUCCGCGGA
1163
2883
AUGCGAGUGCAUCCGCGGA
1163
2901
UCCGCGGAUGCACUCGCAU
1632

2901
ACGCAAGGCCACCCUGGAG
1164
2901
ACGCAAGGCCACCCUGGAG
1164
2919
CUCCAGGGUGGCCUUGCGU
1633

2919
GGAGCUACAGACGGUGCAC
1165
2919
GGAGCUACAGACGGUGCAC
1165
2937
GUGCACCGUCUGUAGCUCC
1634

2937
CUCGGAAGCCCACACCCUC
1166
2937
CUCGGAAGCCCACACCCUC
1166
2955
GAGGGUGUGGGCUUCCGAG
1635

2955
CCUGUAUGGCACGAACCCC
1167
2955
CCUGUAUGGCACGAACCCC
1167
2973
GGGGUUCGUGCCAUACAGG
1636

2973
CCUCAACCGGCAGAAACUG
1168
2973
CCUCAACCGGCAGAAACUG
1168
2991
CAGUUUCUGCCGGUUGAGG
1637

2991
GGACAGUAAGAAACUUCUA
1169
2991
GGACAGUAAGAAACUUCUA
1169
3009
UAGAAGUUUCUUACUGUCC
1638

3009
AGGCUCGCUCGCCUCCGUG
1170
3009
AGGCUCGCUCGCCUCCGUG
1170
3027
CACGGAGGCGAGCGAGCCU
1639

3027
GUUCGUCCGGCUCCCUUGC
1171
3027
GUUCGUCCGGCUCCCUUGC
1171
3045
GCAAGGGAGCCGGACGAAC
1640

3045
CGGUGGUGUUGGGGUGGAC
1172
3045
CGGUGGUGUUGGGGUGGAC
1172
3063
GUCCACCCCAACACCACCG
1641

3063
CAGUGACACCAUAUGGAAC
1173
3063
CAGUGACACCAUAUGGAAC
1173
3081
GUUCCAUAUGGUGUCACUG
1642

3081
CGAGGUGCACUCGGCGGGG
1174
3081
CGAGGUGCACUCGGCGGGG
1174
3099
CCCCGCCGAGUGCACCUCG
1643

3099
GGCAGCCCGCCUGGCUGUG
1175
3099
GGCAGCCCGCCUGGCUGUG
1175
3117
CACAGCCAGGCGGGCUGCC
1644

3117
GGGCUGCGUGGUAGAGCUG
1176
3117
GGGCUGCGUGGUAGAGCUG
1176
3135
CAGCUCUACCACGCAGCCC
1645

3135
GGUCUUCAAGGUGGCCACA
1177
3135
GGUCUUCAAGGUGGCCACA
1177
3153
UGUGGCCACCUUGAAGACC
1646

3153
AGGGGAGCUGAAGAAUGGC
1178
3153
AGGGGAGCUGAAGAAUGGC
1178
3171
GCCAUUCUUCAGCUCCCCU
1647

3171
CUUUGCUGUGGUCCGCCCC
1179
3171
CUUUGCUGUGGUCCGCCCC
1179
3189
GGGGCGGACCACAGCAAAG
1648

3189
CCCUGGACACCAUGCGGAG
1180
3189
CCCUGGACACCAUGCGGAG
1180
3207
CUCCGCAUGGUGUCCAGGG
1649

3207
GGAGAGCACGCCCAUGGGC
1181
3207
GGAGAGCACGCCCAUGGGC
1181
3225
GCCCAUGGGCGUGCUCUCC
1650

3225
CUUUUGCUACUUCAACUCC
1182
3225
CUUUUGCUACUUCAACUCC
1182
3243
GGAGUUGAAGUAGCAAAAG
1651

3243
CGUGGCCGUGGCAGCCAAG
1183
3243
CGUGGCCGUGGCAGCCAAG
1183
3261
CUUGGCUGCCACGGCCACG
1652

3261
GCUUCUGCAGCAGAGGUUG
1184
3261
GCUUCUGCAGCAGAGGUUG
1184
3279
CAACCUCUGCUGCAGAAGC
1653

3279
GAGCGUGAGCAAGAUCCUC
1185
3279
GAGCGUGAGCAAGAUCCUC
1185
3297
GAGGAUCUUGCUCACGCUC
1654

3297
CAUCGUGGACUGGGACGUG
1186
3297
CAUCGUGGACUGGGACGUG
1186
3315
CACGUCCCAGUCCACGAUG
1655

3315
GCACCAUGGAAACGGGACC
1187
3315
GCACCAUGGAAACGGGACC
1187
3333
GGUCCCGUUUCCAUGGUGC
1656

3333
GCAGCAGGCUUUCUACAGC
1188
3333
CCAGCAGGCUUUCUACAGC
1188
3351
GCUGUAGAAAGCCUGCUGG
1657

3351
CGACCCUAGCGUCCUGUAC
1189
3351
CGACCCUAGCGUCCUGUAC
1189
3369
GUACAGGACGCUAGGGUCG
1658

3369
CAUGUCCCUCCACCGCUAC
1190
3369
CAUGUCCCUCCACCGGUAC
1190
3387
GUAGCGGUGGAGGGACAUG
1659

3387
CGACGAUGGGAACUUCUUC
1191
3387
CGACGAUGGGAACUUCUUC
1191
3405
GAAGAAGUUCCCAUCGUCG
1660

3405
CCCAGGCAGCGGGGCUCCU
1192
3405
CCCAGGCAGCGGGGCUCCU
1192
3423
AGGAGCCCCGCUGCCUGGG
1661

3423
UGAUGAGGUGGGCACAGGG
1193
3423
UGAUGAGGUGGGCACAGGG
1193
3441
CCCUGUGCCCACCUCAUCA
1662

3441
GCCCGGCGUGGGUUUCAAC
1194
3441
GCCCGGCGUGGGUUUCAAC
1194
3459
GUUGAAACCCACGCCGGGC
1663

3459
CGUCAACAUGGCUUUCACC
1195
3459
CGUCAACAUGGCUUUCACC
1195
3477
GGUGAAAGCCAUGUUGACG
1664

3477
CGGCGGCCUGGACCCCCCC
1196
3477
CGGCGGCCUGGACCCCCCC
1196
3495
GGGGGGGUCCAGGCCGCCG
1665

3495
CAUGGGAGACGCUGAGUAC
1197
3495
CAUGGGAGACGCUGAGUAC
1197
3513
GUACUCAGCGUCUCCCAUG
1666

3513
CUUGGCGGCCUUCAGAACG
1198
3513
CUUGGCGGCCUUCAGAACG
1198
3531
CGUUCUGAAGGCCGCCAAG
1667

3531
GGUGGUCAUGCCGAUCGCC
1199
3531
GGUGGUCAUGCCGAUCGCC
1199
3549
GGCGAUCGGCAUGACCACC
1668

3549
CAGCGAGUUUGCCCCGGAU
1200
3549
CAGCGAGUUUGCCCCGGAU
1200
3567
AUCCGGGGCAAACUCGCUG
1669

3567
UGUGGUGCUGGUGUCAUCA
1201
3567
UGUGGUGCUGGUGUCAUCA
1201
3585
UGAUGACACCAGCACCACA
1670

3585
AGGCUUCGAUGCCGUGGAG
1202
3585
AGGCUUCGAUGCCGUGGAG
1202
3603
CUCCACGGCAUCGAAGCCU
1671

3603
GGGCCACCCCACCCCUCUU
1203
3603
GGGCCACCCCACCCCUCUU
1203
3621
AAGAGGGGUGGGGUGGCCC
1672

3621
UGGGGGCUACAACCUCUCC
1204
3621
UGGGGGCUACAACCUCUCC
1204
3639
GGAGAGGUUGUAGCCCCCA
1673

3639
CGCCAGAUGCUUCGGGUAC
1205
3639
CGCCAGAUGCUUCGGGUAC
1205
3657
GUACCCGAAGCAUCUGGCG
1674

3657
CCUGACGAAGCAGCUGAUG
1206
3657
CCUGACGAAGCAGCUGAUG
1206
3675
CAUCAGCUGCUUCGUCAGG
1675

3675
GGGCCUGGCUGGCGGCCGG
1207
3675
GGGCCUGGCUGGCGGCCGG
1207
3693
CCGGCCGCCAGCCAGGCCC
1676

3693
GAUUGUCCUGGCCCUCGAG
1208
3693
GAUUGUCCUGGCCCUCGAG
1208
3711
CUCGAGGGCCAGGACAAUC
1677

3711
GGGAGGCCACGACCUGACC
1209
3711
GGGAGGCCACGACCUGACC
1209
3729
GGUCAGGUCGUGGCCUCCC
1678

3729
CGCCAUUUGCGACGCCUCG
1210
3729
CGCCAUUUGCGACGCCUCG
1210
3747
CGAGGCGUCGCAAAUGGCG
1679

3747
GGAAGCAUGUGUUUCUGCC
1211
3747
GGAAGCAUGUGUUUCUGCC
1211
3765
GGCAGAAACACAUGCUUCC
1680

3765
CUUGCUGGGAAACGAGCUU
1212
3765
CUUGCUGGGAAACGAGCUU
1212
3783
AAGCUCGUUUCCCAGCAAG
1681

3783
UGAUCCUCUCCCAGAAAAG
1213
3783
UGAUCCUCUCCCAGAAAAG
1213
3801
CUUUUCUGGGAGAGGAUCA
1682

3801
GGUUUUACAGCAAAGACCC
1214
3801
GGUUUUACAGCAAAGACCC
1214
3819
GGGUCUUUGCUGUAAAACC
1683

3819
CAAUGCAAACGCUGUCCGU
1215
3819
CAAUGCAAACGCUGUCCGU
1215
3837
ACGGACAGCGUUUGCAUUG
1684

3837
UUCCAUGGAGAAAGUCAUG
1216
3837
UUCCAUGGAGAAAGUCAUG
1216
3855
CAUGACUUUCUCCAUGGAA
1685

3855
GGAGAUCCACAGCAAGUAC
1217
3855
GGAGAUCCACAGCAAGUAC
1217
3873
GUACUUGCUGUGGAUCUCC
1686

3873
CUGGCGCUGCCUGCAGCGC
1218
3873
CUGGCGCUGCCUGCAGCGC
1218
3891
GCGCUGCAGGCAGCGCCAG
1687

3891
CACAACCUCCACAGCGGGG
1219
3891
CACAACGUCCACAGCGGGG
1219
3909
CCCCGCUGUGGAGGUUGUG
1688

3909
GCGUUCUCUGAUCGAGGCU
1220
3909
GCGUUCUCUGAUCGAGGCU
1220
3927
AGGCUCGAUCAGAGAACGC
1689

3927
UCAGACUUGCGAGAACGAA
1221
3927
UCAGACUUGCGAGAACGAA
1221
3945
UUCGUUCUCGCAAGUCUGA
1690

3945
AGAAGCCGAGACGGUCACC
1222
3945
AGAAGCCGAGACGGUCACC
1222
3963
GGUGACCGUCUCGGCUUCU
1691

3963
CGCCAUGGCCUCGCUGUCC
1223
3963
CGCCAUGGCCUCGCUGUCC
1223
3981
GGACAGCGAGGCCAUGGCG
1692

3981
CGUGGGCGUGAAGCCCGCC
1224
3981
CGUGGGCGUGAAGCCCGCC
1224
3999
GGCGGGCUUCACGCCCACG
1693

3999
CGAAAAGAGACCAGAUGAG
1225
3999
CGAAAAGAGACCAGAUGAG
1225
4017
CUCAUCUGGUCUCUUUUCG
1694

4017
GGAGCCCAUGGAAGAGGAG
1226
4017
GGAGCCCAUGGAAGAGGAG
1226
4035
CUCCUCUUCCAUGGGCUCC
1695

4035
GCCGCCCCUGUAGCACUCC
1227
4035
GCCGCCCCUGUAGCACUCC
1227
4053
GGAGUGCUACAGGGGCGGC
1696

4053
CCUCGAAGCUGCUGUUCUC
1228
4053
CCUCGAAGCUGCUGUUCUC
1228
4071
GAGAACAGCAGCUUCGAGG
1697

4071
CUUGUCUGUCUGUCUCUGU
1229
4071
CUUGUCUGUCUGUCUCUGU
1229
4089
ACAGAGACAGACAGACAAG
1698

4089
UCUUGAAGCUCAGCCAAGA
1230
4089
UCUUGAAGCUCAGCCAAGA
1230
4107
UCUUGGCUGAGCUUCAAGA
1699

4107
AAACUUUCCCGUGUCACGC
1231
4107
AAACUUUCCCGUGUCACGC
1231
4125
GCGUGACACGGGAAAGUUU
1700

4125
CCUGCGUCCCACCGUGGGG
1232
4125
CCUGCGUCCCACCGUGGGG
1232
4143
CCCCACGGUGGGACGCAGG
1701

4143
GCUCUCUUGGAGCACCCAG
1233
4143
GCUCUCUUGGAGCACCCAG
1233
4161
CUGGGUGCUCCAAGAGAGC
1702

4161
GGGACACCCAGCGUGCAAC
1234
4161
GGGACACCCAGCGUGCAAC
1234
4179
GUUGCACGCUGGGUGUCCC
1703

4179
CAGCCACGGGAAGCCUUUC
1235
4179
CAGCCACGGGAAGCCUUUC
1235
4197
GAAAGGCUUCCCGUGGCUG
1704

4197
CUGCCGCCCAGGCCCACAG
1236
4197
CUGCCGCCCAGGCCCACAG
1236
4215
CUGUGGGCCUGGGCGGCAG
1705

4215
GGUCUCGAGACGCACAUGC
1237
4215
GGUCUCGAGACGCACAUGC
1237
4233
GCAUGUGCGUCUCGAGACC
1706

4233
CACGCCUGGGCGUGGCAGC
1238
4233
CACGCCUGGGCGUGGCAGC
1238
4251
GCUGCCACGCCCAGGCGUG
1707

4251
CCUCACAGGGAACACGGGA
1239
4251
CCUCACAGGGAACACGGGA
1239
4269
UCCCGUGUUCCCUGUGAGG
1708

4269
ACAGACGCCGGCGACGCGC
1240
4269
ACAGACGCCGGCGACGCGC
1240
4287
GCGCGUCGCCGGCGUCUGU
1709

4287
CAGACACACGGACACGCGG
1241
4287
CAGACACACGGACACGCGG
1241
4305
CCGCGUGUCCGUGUGUCUG
1710

4305
GAAGCCAAGCACACUCUGG
1242
4305
GAAGCCAAGCACACUCUGG
1242
4323
CCAGAGUGUGCUUGGCUUC
1711

4323
GCGGGUCCCGCAAGGGACG
1243
4323
GCGGGUCCCGCAAGGGACG
1243
4341
CGUCCCUUGCGGGACCCGC
1712

4341
GCCGUGGAAGAAAGGAGCC
1244
4341
GCCGUGGAAGAAAGGAGCC
1244
4359
GGCUCCUUUCUUCCACGGC
1713

4359
CUGUGGCAACAGGCGGCCG
1245
4359
CUGUGGCAACAGGCGGCCG
1245
4377
CGGCCGCCUGUUGCCACAG
1714

4377
GAGCUGCCGAAUUCAGUUG
1246
4377
GAGCUGCCGAAUUCAGUUG
1246
4395
CAACUGAAUUCGGCAGCUC
1715

4395
GACACGAGGCACAGAAAAC
1247
4395
GACACGAGGCACAGAAAAC
1247
4413
GUUUUCUGUGCCUCGUGUC
1716

4413
CAAAUAUCAAAGAUCUAAU
1248
4413
CAAAUAUCAAAGAUCUAAU
1248
4431
AUUAGAUCUUUGAUAUUUG
1717

4431
UAAUACAAAACAAACUUGA
1249
4431
UAAUACAAAACAAACUUGA
1249
4449
UCAAGUUUGUUUUGUAUUA
1718

4449
AUUAAAACUGGUGCUUAAA
1250
4449
AUUAAAACUGGUGCUUAAA
1250
4467
UUUAAGCACCAGUUUUAAU
1719

4467
AGUUUAUUACCCACAACUC
1251
4467
AGUUUAUUACCCACAACUC
1251
4485
GAGUUGUGGGUAAUAAACU
1720

4485
CCACAGUCUCUGUGUAAAC
1252
4485
CCACAGUCUCUGUGUAAAC
1252
4503
GUUUACACAGAGACUGUGG
1721

4503
CCACUCGACUCAUCUUGUA
1253
4503
CCACUCGACUCAUCUUGUA
1253
4521
UACAAGAUGAGUCGAGUGG
1722

4521
AGCUUAUUUUUUUUUUAAA
1254
4521
AGCUUAUUUUUUUUUUAAA
1254
4539
UUUAAAAAAAAAAUAAGCU
1723

4539
AGAGGACGUUUUCUACGGC
1255
4539
AGAGGACGUUUUCUACGGC
1255
4557
GCCGUAGAAAACGUCCUCU
1724

4557
CUGUGGCCCGCCUCUGUGA
1256
4557
CUGUGGCCCGCCUCUGUGA
1256
4575
UCACAGAGGCGGGCCACAG
1725

4575
AACCAUAGCGGUGUGCGGC
1257
4575
AACCAUAGCGGUGUGCGGC
1257
4593
GCCGCACACCGCUAUGGUU
1726

4593
CGGGGGGUCUGCACCCGGG
1258
4593
CGGGGGGUCUGCACCCGGG
1258
4611
CCCGGGUGCAGACCCCCCG
1727

4611
GUGGGGGACAGAGGGACCU
1259
4611
GUGGGGGACAGAGGGACCU
1259
4629
AGGUCCCUCUGUCCCCCAC
1728

4629
UUUAAAGAAAACAAAACUG
1260
4629
UUUAAAGAAAACAAAACUG
1260
4647
CAGUUUUGUUUUCUUUAAA
1729

4647
GGACAGAAACAGGAAUGUG
1261
4647
GGACAGAAACAGGAAUGUG
1261
4665
CACAUUCCUGUUUCUGUCC
1730

4665
GAGCUGGGGGAGCUGGCUU
1262
4665
GAGCUGGGGGAGCUGGCUU
1262
4683
AAGCCAGCUCCCCCAGCUC
1731

4683
UGAGUUUCUCAAAAGCCAU
1263
4683
UGAGUUUCUCAAAAGCCAU
1263
4701
AUGGCUUUUGAGAAACUCA
1732

4701
UCGGAAGAUGCGAGUUUGU
1264
4701
UCGGAAGAUGCGAGUUUGU
1264
4719
ACAAACUCGCAUCUUCCGA
1733

4719
UGCCUUUUUUUUUAUUGCU
1265
4719
UGCCUUUUUUUUUAUUGCU
1265
4737
AGCAAUAAAAAAAAAGGCA
1734

4737
UCUGGUGGAUUUUUGUGGC
1266
4737
UCUGGUGGAUUUUUGUGGC
1266
4755
GCCACAAAAAUCCACCAGA
1735

4755
CUGGGUUUUCUGAAGUCUG
1267
4755
CUGGGUUUUCUGAAGUCUG
1267
4773
CAGACUUCAGAAAACCCAG
1736

4773
GAGGAACAAUGCCUUAAGA
1268
4773
GAGGAACAAUGCCUUAAGA
1268
4791
UCUUAAGGCAUUGUUCCUC
1737

4791
AAAAAACAAACAGCAGGAA
1269
4791
AAAAAACAAACAGCAGGAA
1269
4809
UUCCUGCUGUUUGUUUUUU
1738

4809
AUCGGUGGGACAGUUUCCU
1270
4809
AUCGGUGGGACAGUUUCCU
1270
4827
AGGAAACUGUCCCACCGAU
1739

4827
UGUGGCCAGCCGAGCCUGG
1271
4827
UGUGGCCAGCCGAGCCUGG
1271
4845
CCAGGCUCGGCUGGCCACA
1740

4845
GCAGUGCUGGCACCGCGAG
1272
4845
GCAGUGCUGGCACCGCGAG
1272
4863
CUCGCGGUGCCAGCACUGC
1741

4863
GCUGGCCUGACGCCUCAAG
1273
4863
GCUGGCCUGACGCCUCAAG
1273
4881
CUUGAGGCGUCAGGCCAGC
1742

4881
GCACGGGCACCAGCCGUCA
1274
4881
GGACGGGCACCAGCCGUCA
1274
4899
UGACGGCUGGUGCCCGUGC
1743

4899
AUCUCCGGGGCCAGGGGCU
1275
4899
AUCUCCGGGGCCAGGGGCU
1275
4917
AGCCCCUGGCCCCGGAGAU
1744

4917
UGCAGCCCGGCGGUCCCUG
1276
4917
UGCAGCCCGGCGGUCCCUG
1276
4935
CAGGGACCGCCGGGCUGCA
1745

4935
GUUUUGCUUUAUUGCUGUU
1277
4935
GUUUUGCUUUAUUGCUGUU
1277
4953
AACAGCAAUAAAGCAAAAC
1746

4953
UUAAGAAAAAUGGAGGUAG
1278
4953
UUAAGAAAAAUGGAGGUAG
1278
4971
CUACCUCCAUUUUUCUUAA
1747

4971
GUUCCAAAAAAGUGGCAAA
1279
4971
GUUCCAAAAAAGUGGCAAA
1279
4989
UUUGCCACUUUUUUGGAAC
1748

4989
AUCCCGUUGGAGGUUUUGA
1280
4989
AUCCCGUUGGAGGUUUUGA
1280
5007
UCAAAACCUCCAACGGGAU
1749

5007
AAGUCCAACAAAUUUUAAA
1281
5007
AAGUCCAACAAAUUUUAAA
1281
5025
UUUAAAAUUUGUUGGACUU
1750

5025
ACGAAUCCAAAGUGUUCUC
1282
5025
ACGAAUCCAAAGUGUUCUC
1282
5043
GAGAACACUUUGGAUUCGU
1751

5043
CACAGGUCACAUACGAUUG
1283
5043
CACACGUCACAUACGAUUG
1283
5061
CAAUCGUAUGUGACGUGUG
1752

5061
GAGCAUCUCCAUCUGGUCG
1284
5061
GAGCAUCUCCAUCUGGUCG
1284
5079
CGACCAGAUGGAGAUGCUC
1753

5079
GUGAAGCAUGUGGUAGGCA
1285
5079
GUGAAGCAUGUGGUAGGCA
1285
5097
UGCCUACCACAUGCUUCAC
1754

5097
ACACUUGCAGUGUUACGAU
1286
5097
ACACUUGCAGUGUUACGAU
1286
5115
AUCGUAACACUGCAAGUGU
1755

5115
UCGGAAUGCUUUUUAUUAA
1287
5115
UCGGAAUGCUUUUUAUUAA
1287
5133
UUAAUAAAAAGCAUUCCGA
1756

5133
AAAGCAAGUAGCAUGAAGU
1288
5133
AAAGCAAGUAGCAUGAAGU
1288
5151
ACUUCAUGCUACUUGCUUU
1757

5151
UAUUGCUUAAAUUUUAGGU
1289
5151
UAUUGCUUAAAUUUUAGGU
1289
5169
ACCUAAAAUUUAAGCAAUA
1758

5169
UAUAAAUAAAUAUAUAUAU
1290
5169
UAUAAAUAAAUAUAUAUAU
1290
5187
AUAUAUAUAUUUAUUUAUA
1759

5187
UGUAUAAUAUAUAUUCCAA
1291
5187
UGUAUAAUAUAUAUUCCAA
1291
5205
UUGGAAUAUAUAUUAUACA
1760

5205
AUGUAUUCCAAGCUAAGAA
1292
5205
AUGUAUUCCAAGCUAAGAA
1292
5223
UUCUUAGCUUGGAAUACAU
1761

5223
AACUUACUUGAUUCUUAUG
1293
5223
AACUUACUUGAUUCUUAUG
1293
5241
CAUAAGAAUCAAGUAAGUU
1762

5241
GAAAUCUUGAUAAAAUAUU
1294
5241
GAAAUCUUGAUAAAAUAUU
1294
5259
AAUAUUUUAUCAAGAUUUC
1763

5259
UUAUAAUGCAUUUAUAGAA
1295
5259
UUAUAAUGCAUUUAUAGAA
1295
5277
UUCUAUAAAUGCAUUAUAA
1764

5277
AAAAGUAUAUAUAUAUAUA
1296
5277
AAAAGUAUAUAUAUAUAUA
1296
5295
UAUAUAUAUAUAUACUUUU
1765

5295
AUAAAAUGAAUGCAGAUUG
1297
5295
AUAAAAUGAAUGCAGAUUG
1297
5313
CAAUCUGCAUUCAUUUUAU
1766

5313
GCGAAGGUCCCUGCAAAUG
1298
5313
GCGAAGGUCCCUGCAAAUG
1298
5331
CAUUUGCAGGGACCUUCGC
1767

5331
GGAUGGCUUGUGAAUUUGC
1299
5331
GGAUGGCUUGUGAAUUUGC
1299
5349
GCAAAUUCACAAGCCAUCC
1768

5349
CUCUCAAGGUGCUUAUGGA
1300
5349
CUCUCAAGGUGCUUAUGGA
1300
5367
UCCAUAAGCACCUUGAGAG
1769

5367
AAAGGGAUCCUGAUUGAUU
1301
5367
AAAGGGAUCCUGAUUGAUU
1301
5385
AAUCAAUCAGGAUCCCUUU
1770

5385
UGAAAUUCAUGUUUUCUCA
1302
5385
UGAAAUUCAUGUUUUCUCA
1302
5403
UGAGAAAACAUGAAUUUCA
1771

5403
AAGCUCCAGAUUGGCUAGA
1303
5403
AAGCUCCAGAUUGGCUAGA
1303
5421
UCUAGCCAAUCUGGAGCUU
1772

5421
AUUUCAGAUCGCCAACACA
1304
5421
AUUUCAGAUCGCCAACACA
1304
5439
UGUGUUGGCGAUCUGAAAU
1773

5439
AUUCGCCACUGGGCAACUA
1305
5439
AUUCGCCACUGGGCAACUA
1305
5457
UAGUUGCCCAGUGGCGAAU
1774

5457
ACCCUACAAGUUUGUACUU
1306
5457
ACCCUACAAGUUUGUACUU
1306
5475
AAGUACAAACUUGUAGGGU
1775

5475
UUCAUUUUAAUUAUUUUCU
1307
5475
UUCAUUUUAAUUAUUUUCU
1307
5493
AGAAAAUAAUUAAAAUGAA
1776

5493
UAACAGAACCGCUCCCGUC
1308
5493
UAACAGAACCGCUCCCGUC
1308
5511
GACGGGAGCGGUUCUGUUA
1777

5511
CUCCAAGCCUUCAUGCACA
1309
5511
CUCCAAGCCUUCAUGCACA
1309
5529
UGUGCAUGAAGGCUUGGAG
1778

5529
AUAUGUACCUAAUGAGUUU
1310
5529
AUAUGUACCUAAUGAGUUU
1310
5547
AAACUCAUUAGGUACAUAU
1779

5547
UUUAUAGCAAAGAAUAUAA
1311
5547
UUUAUAGCAAAGAAUAUAA
1311
5565
UUAUAUUCUUUGCUAUAAA
1780

5565
AAUUUGCUGUUGAUUUUUG
1312
5565
AAUUUGCUGUUGAUUUUUG
1312
5583
CAAAAAUCAACAGCAAAUU
1781

5583
GUAUGAAUUUUUUCACAAA
1313
5583
GUAUGAAUUUUUUCACAAA
1313
5601
UUUGUGAAAAAAUUCAUAC
1782

5601
AAAGAUCCUGAAUAAGCAU
1314
5601
AAAGAUCCUGAAUAAGCAU
1314
5619
AUGCUUAUUCAGGAUCUUU
1783

5619
UUGUUUUAUGAAUUUUACA
1315
5619
UUGUUUUAUGAAUUUUACA
1315
5637
UGUAAAAUUCAUAAAACAA
1784

5637
AUUUUUCCUCACCAUUUAG
1316
5637
AUUUUUCCUCACCAUUUAG
1316
5655
CUAAAUGGUGAGGAAAAAU
1785

5655
GCAAUUUUCUGAAUGGUAA
1317
5655
GCAAUUUUCUGAAUGGUAA
1317
5673
UUACCAUUCAGAAAAUUGC
1786

5673
AUAAUGUCUAAAUCUUUUU
1318
5673
AUAAUGUCUAAAUCUUUUU
1318
5691
AAAAAGAUUUAGACAUUAU
1787

5691
UCCUUUCUGAAUUCUUGCU
1319
5691
UCCUUUCUGAAUUCUUGCU
1319
5709
AGCAAGAAUUCAGAAAGGA
1788

5709
UUGUACAUUUUUUUUUACC
1320
5709
UUGUACAUUUUUUUUUACC
1320
5727
GGUAAAAAAAAAUGUACAA
1789

5727
CUUUCAAAGGUUUUUAAUU
1321
5727
CUUUCAAAGGUUUUUAAUU
1321
5745
AAUUAAAAACCUUUGAAAG
1790

5745
UAUUUUUGUUUUUAUUUUU
1322
5745
UAUUUUUGUUUUUAUUUUU
1322
5763
AAAAAUAAAAACAAAAAUA
1791

5763
UGUACGAUGAGUUUUCUGC
1323
5763
UGUACGAUGAGUUUUCUGC
1323
5781
GCAGAAAACUCAUCGUACA
1792

5781
CAGCGUACAGAAUUGUUGC
1324
5781
CAGCGUACAGAAUUGUUGC
1324
5799
GCAACAAUUCUGUACGCUG
1793

5799
CUGUCAGAUUCUAUUUUCA
1325
5799
CUGUCAGAUUCUAUUUUCA
1325
5817
UGAAAAUAGAAUCUGACAG
1794

5817
AGAAAGUGAGAGGAGGGAC
1326
5817
AGAAAGUGAGAGGAGGGAC
1326
5835
GUCCCUCCUCUCACUUUCU
1795

5835
CCGUAGGUCUUUUCGGAGU
1327
5835
CCGUAGGUCUUUUCGGAGU
1327
5853
ACUCCGAAAAGACCUACGG
1796

5853
UGACACCAACGAUUGUGUC
1328
5853
UGACACCAACGAUUGUGUC
1328
5871
GACACAAUCGUUGGUGUCA
1797

5871
CUUUCCUGGUCUGUCCUAG
1329
5871
CUUUCCUGGUCUGUCCUAG
1329
5889
CUAGGACAGACCAGGAAAG
1798

5889
GGAGCUGUAUAAAGAAGCC
1330
5889
GGAGCUGUAUAAAGAAGCC
1330
5907
GGCUUCUUUAUACAGCUCC
1799

5907
CCAGGGGCUCUUUUUAACU
1331
5907
CCAGGGGCUCUUUUUAACU
1331
5925
AGUUAAAAAGAGCCCCUGG
1800

5925
UUUCAACACUAGUAGUAUU
1332
5925
UUUCAACACUAGUAGUAUU
1332
5943
AAUACUACUAGUGUUGAAA
1801

5943
UACGAGGGGUGGUGUGUUU
1333
5943
UACGAGGGGUGGUGUGUUU
1333
5961
AAACACACCACCCCUCGUA
1802

5961
UUUCCCCUCCGUGGCAAGG
1334
5961
UUUCCCCUCCGUGGCAAGG
1334
5979
CCUUGCCACGGAGGGGAAA
1803

5979
GGCAGGGAGGGUUGCUUAG
1335
5979
GGCAGGGAGGGUUGCUUAG
1335
5997
CUAAGCAACCCUCCCUGCC
1804

5997
GGAUGCCCGGCCACCCUGG
1336
5997
GGAUGCCCGGCCACCCUGG
1336
6015
CCAGGGUGGCCGGGCAUCC
1805

6015
GGAGGCUUGCCAGAUGCCG
1337
6015
GGAGGCUUGCCAGAUGCCG
1337
6033
CGGCAUCUGGCAAGCCUCC
1806

6033
GGGGGCAGUCAGCAUUAAU
1338
6033
GGGGGCAGUCAGCAUUAAU
1338
6051
AUUAAUGCUGACUGCCCCC
1807

6051
UGAAACUCAUGUUUAAACU
1339
6051
UGAAACUCAUGUUUAAACU
1339
6069
AGUUUAAACAUGAGUUUCA
1808

6069
UUCUCUGACCACAUCGUCA
1340
6069
UUCUCUGACCACAUCGUCA
1340
6087
UGACGAUGUGGUCAGAGAA
1809

6O87
AGGAUAGAAUUCUAACUUG
1341
6087
AGGAUAGAAUUCUAACUUG
1341
6105
CAAGUUAGAAUUCUAUCCU
1810

6105
GAGUUUUCCAAAGACCUUU
1342
6105
GAGUUUUCCAAAGACCUUU
1342
6123
AAAGGUCUUUGGAAAACUC
1811

6123
UUGAGCAUGUCAGCAAUGC
1343
6123
UUGAGCAUGUCAGCAAUGC
1343
6141
GCAUUGCUGACAUGCUCAA
1812

6141
CAUGGGGCACACGUGGGGC
1344
6141
CAUGGGGCACACGUGGGGC
1344
6159
GCCCCACGUGUGCCCCAUG
1813

6159
CUCUUUACCCACUUGGGUU
1345
6159
CUCUUUACCCACUUGGGUU
1345
6177
AACCCAAGUGGGUAAAGAG
1814

6177
UUUUCCACUGCAGCCACGU
1346
6177
UUUUCCACUGCAGCCACGU
1346
6195
ACGUGGCUGCAGUGGAAAA
1815

6195
UGGCCAGCCCUGGAUUUUG
1347
6195
UGGCCAGCCCUGGAUUUUG
1347
6213
CAAAAUCCAGGGCUGGCCA
1816

6213
GGAGCCUGUGGCUGCAAGG
1348
6213
GGAGCCUGUGGCUGCAAGG
1348
6231
CCUUGCAGCCACAGGCUCC
1817

6231
GAACCCAGGGACCCUUGUU
1349
6231
GAACCCAGGGACCCUUGUU
1349
6249
AACAAGGGUCCCUGGGUUC
1818

6249
UGCCUGGUGAACCUGCAGG
1350
6249
UGCCUGGUGAACCUGCAGG
1350
6267
CCUGCAGGUUCACCAGGCA
1819

6267
GGAGGGUAUGAUUGCCUGA
1351
6267
GGAGGGUAUGAUUGCCUGA
1351
6285
UCAGGCAAUCAUACCCUCC
1820

6285
ACCAGGACAGCCAGUCUUU
1352
6285
ACCAGGACAGCCAGUCUUU
1352
6303
AAAGACUGGCUGUCCUGGU
1821

6303
UACUCUUUUUCUCUUCAAC
1353
6303
UACUCUUUUUCUCUUCAAC
1353
6321
GUUGAAGAGAAAAAGAGUA
1822

6321
CAGUAACUGACAGUCACGU
1354
6321
CAGUAACUGACAGUCACGU
1354
6339
ACGUGACUGUCAGUUACUG
1823

6339
UUUUACUGGUAACUUAUUU
1355
6339
UUUUACUGGUAACUUAUUU
1355
6357
AAAUAAGUUACCAGUAAAA
1824

6357
UUCCAGCACAUGAAGCCAC
1356
6357
UUCCAGCACAUGAAGCCAC
1356
6375
GUGGCUUCAUGUGCUGGAA
1825

6375
CCAGUUUCAUUCCAAAGUG
1357
6375
CCAGUUUCAUUCCAAAGUG
1357
6393
CACUUUGGAAUGAAACUGG
1826

6393
GUAUAUUGGGUUCAGACUU
1358
6393
GUAUAUUGGGUUCAGACUU
1358
6411
AAGUCUGAACCCAAUAUAC
1827

6411
UGGGGGCAGAAGUUCAGAC
1359
6411
UGGGGGCAGAAGUUCAGAC
1359
6429
GUCUGAACUUCUGCCCCCA
1828

6429
CACACCGUGCUCAGGAGGG
1360
6429
CACACCGUGCUCAGGAGGG
1360
6447
CCCUCCUGAGCACGGUGUG
1829

6447
GACCCAGAGCCGAGUUUCG
1361
6447
GACCCAGAGCCGAGUUUCG
1361
6465
CGAAACUCGGCUCUGGGUC
1830

6465
GGAGUUUGGUAAAGUUUAC
1362
6465
GGAGUUUGGUAAAGUUUAC
1362
6483
GUAAACUUUACCAAACUCC
1831

6483
CAGGGUAGCUUCUGAAAUU
1363
6483
CAGGGUAGCUUCUGAAAUU
1363
6501
AAUUUCAGAAGCUACCCUG
1832

6501
UAACUCAAACUUUUGACCA
1364
6501
UAACUCAAACUUUUGACCA
1364
6519
UGGUCAAAAGUUUGAGUUA
1833

6519
AAAUGAGUGCAGAUUCUUG
1365
6519
AAAUGAGUGCAGAUUCUUG
1365
6537
CAAGAAUCUGCACUCAUUU
1834

6537
GGAUUCACUUGGUCACUGG
1366
6537
GGAUUCACUUGGUCACUGG
1366
6555
CCAGUGACCAAGUGAAUCC
1835

6555
GGCUGCUGAUGGUCAGCUC
1367
6555
GGCUGCUGAUGGUCAGCUC
1367
6573
GAGCUGACCAUCAGCAGCC
1836

6573
CUGAGACAGUGGUUUGAGA
1368
6573
CUGAGACAGUGGUUUGAGA
1368
6591
UCUCAAACCACUGUCUCAG
1837

6591
AGCAGGCAGAACGGUCUUG
1369
6591
AGCAGGCAGAACGGUCUUG
1369
6609
CAAGACCGUUCUGCCUGCU
1838

6609
GGGACUUGUUUGACUUUCC
1370
6609
GGGACUUGUUUGACUUUCC
1370
6627
GGAAAGUCAAACAAGUCCC
1839

6627
CCCUCCCUGGUGGCCACUC
1371
6627
CCCUCCCUGGUGGCCACUC
1371
6645
GAGUGGCCACCAGGGAGGG
1840

6645
CUUUGCUCUGAAGCCCAGA
1372
6645
CUUUGCUCUGAAGCCCAGA
1372
6663
UCUGGGCUUCAGAGCAAAG
1841

6663
AUUGGCAAGAGGAGCUGGU
1373
6663
AUUGGCAAGAGGAGCUGGU
1373
6681
ACCAGCUCCUCUUGCCAAU
1842

6681
UCCAUUCCCCAUUCAUGGC
1374
6681
UCCAUUCCCCAUUCAUGGC
1374
6699
GCCAUGAAUGGGGAAUGGA
1843

6699
CACAGAGCAGUGGCAGGGC
1375
6699
CACAGAGCAGUGGCAGGGC
1375
6717
GCCCUGCCACUGCUCUGUG
1844

6717
CCCAGCUAGCAGGCUCUUC
1376
6717
CCCAGCUAGCAGGCUCUUC
1376
6735
GAAGAGCCUGCUAGCUGGG
1845

6735
CUGGCCUCCUUGGCCUCAU
1377
6735
CUGGCCUCCUUGGCCUCAU
1377
6753
AUGAGGCCAAGGAGGCCAG
1846

6753
UUCUCUGCAUAGCCCUCUG
1378
6753
UUCUCUGCAUAGCCCUCUG
1378
6771
CAGAGGGCUAUGCAGAGAA
1847

6771
GGGGAUCCUGCCACCUGCC
1379
6771
GGGGAUCCUGCCACCUGCC
1379
6789
GGCAGGUGGCAGGAUCCCC
1848

6789
CCUCUUACCCCGCCGUGGC
1380
6789
CCUCUUACCCCGCCGUGGC
1380
6807
GCCACGGCGGGGUAAGAGG
1849

6807
CUUAUGGGGAGGAAUGCAU
1381
6807
CUUAUGGGGAGGAAUGGAU
1381
6825
AUGCAUUCCUCCCCAUAAG
1850

6825
UCAUCUCACUUUUUUUUUU
1382
6825
UCAUCUCACUUUUUUUUUU
1382
6843
AAAAAAAAAAGUGAGAUGA
1851

6843
UUAAGCAGAUGAUGGGAUA
1383
6843
UUAAGCAGAUGAUGGGAUA
1383
6861
UAUCCCAUCAUCUGCUUAA
1852

6861
AACAUGGACUGCUCAGUGG
1384
6861
AACAUGGACUGCUCAGUGG
1384
6879
CCACUGAGCAGUCCAUGUU
1853

6879
GCCAGGUUAUCAGUGGGGG
1385
6879
GCCAGGUUAUCAGUGGGGG
1385
6897
CCCCCACUGAUAACCUGGC
1854

6897
GGACUUAAUUCUAAUCUCA
1386
6897
GGACUUAAUUCUAAUCUCA
1386
6915
UGAGAUUAGAAUUAAGUCC
1855

6915
AUUCAAAUGGAGACGCCCU
1387
6915
AUUCAAAUGGAGACGCCCU
1387
6933
AGGGCGUCUCCAUUUGAAU
1856

6933
UCUGCAAAGGCCUGGCAGG
1388
6933
UCUGCAAAGGCCUGGCAGG
1388
6951
CCUGCCAGGCCUUUGCAGA
1857

6951
GGGGAGGCACGUUUCAUCU
1389
6951
GGGGAGGCACGUUUCAUCU
1389
6969
AGAUGAAACGUGCCUCCCC
1858

6969
UGUCAGCUCACUCCAGGUU
1390
6969
UGUCAGCUCACUCCAGCUU
1390
6987
AAGCUGGAGUGAGCUGACA
1859

6987
UCACAAAUGUGCUGAGAGC
1391
6987
UCACAAAUGUGCUGAGAGC
1391
7005
GCUCUCAGCACAUUUGUGA
1860

7005
CAUUACUGUGUAGCCUUUU
1392
7005
CAUUACUGUGUAGCCUUUU
1392
7023
AAAAGGCUACACAGUAAUG
1861

7023
UCUUUGAAGACACACUCGG
1393
7023
UCUUUGAAGACACACUCGG
1393
7041
CCGAGUGUGUCUUCAAAGA
1862

7041
GCUCUUCUCCACAGCAAGC
1394
7041
GCUCUUCUCCACAGCAAGC
1394
7059
GCUUGCUGUGGAGAAGAGC
1863

7059
CGUCCAGGGCAGAUGGCAG
1395
7059
CGUCCAGGGCAGAUGGCAG
1395
7077
CUGCCAUCUGCCCUGGACG
1864

7077
GAGGAUCUGCCUCGGCGUC
1396
7077
GAGGAUCUGCCUCGGCGUC
1396
7095
GACGCCGAGGCAGAUCCUC
1865

7095
CUGCAGGCGGGACGACGUC
1397
7095
CUGCAGGCGGGACCACGUC
1397
7113
GACGUGGUCCCGCCUGCAG
1866

7113
CAGGGAGGGUUCCUUCAUG
1398
7113
CAGGGAGGGUUCCUUCAUG
1398
7131
CAUGAAGGAACCCUCCCUG
1867

7131
GUGUUCUCCCUGUGGGUCC
1399
7131
GUGUUCUCCCUGUGGGUCC
1399
7149
GGACCCACAGGGAGAACAC
1868

7149
CUUGGACCUUUAGCCUUUU
1400
7149
CUUGGACCUUUAGCCUUUU
1400
7167
AAAAGGCUAAAGGUCCAAG
1869

7167
UUCUUCCUUUGCAAAGGCC
1401
7167
UUCUUCCUUUGCAAAGGCC
1401
7185
GGCCUUUGCAAAGGAAGAA
1870

7185
CUUGGGGGCACUGGCUGGG
1402
7185
CUUGGGGGCACUGGCUGGG
1402
7203
CCCAGCCAGUGCCCCCAAG
1871

7203
GAGUCAGCAAGCGAGCACU
1403
7203
GAGUCAGCAAGCGAGCACU
1403
7221
AGUGCUCGCUUGCUGACUC
1872

7221
UUUAUAUCCCUUUGAGGGA
1404
7221
UUUAUAUCCCUUUGAGGGA
1404
7239
UCCCUCAAAGGGAUAUAAA
1873

7239
AAACCCUGAUGACGCCACU
1405
7239
AAACCCUGAUGACGCCACU
1405
7257
AGUGGCGUCAUCAGGGUUU
1874

7257
UGGGCCUCUUGGCGUCUGC
1406
7257
UGGGCCUCUUGGCGUCUGC
1406
7275
GCAGACGCCAAGAGGCCCA
1875

7275
CCCUGCCCUCGCGGCUUCC
1407
7275
CCCUGCCCUCGCGGCUUCC
1407
7293
GGAAGCCGCGAGGGCAGGG
1876

7293
CCGCCGUGCCGCAGCGUGC
1408
7293
CCGCCGUGCCGCAGCGUGC
1408
7311
GCACGCUGCGGCACGGCGG
1877

7311
CCCACGUGCCCACGCCCCA
1409
7311
CCCACGUGCCCACGCCCCA
1409
7329
UGGGGCGUGGGCACGUGGG
1878

7329
ACCAGCAGGCGGCUGUCCC
1410
7329
ACCAGCAGGCGGCUGUCCC
1410
7347
GGGACAGCCGCCUGCUGGU
1879

7347
CGGAGGCCGUGGCCCGCUG
1411
7347
CGGAGGCCGUGGCCCGCUG
1411
7365
CAGCGGGCCACGGCCUCCG
1880

7365
GGGACUGGCCGCCCCUCCC
1412
7365
GGGACUGGCCGCCCCUCCC
1412
7383
GGGAGGGGCGGCCAGUCCC
1881

7383
CCAGCGUCCCAGGGCUCUG
1413
7383
CCAGCGUCCCAGGGCUCUG
1413
7401
CAGAGCCCUGGGACGCUGG
1882

7401
GGUUCUGGAGGGCCACUUU
1414
7401
GGUUCUGGAGGGCCACUUU
1414
7419
AAAGUGGCCCUCCAGAACC
1883

7419
UGUCAAGGUGUUUCAGUUU
1415
7419
UGUCAAGGUGUUUCAGUUU
1415
7437
AAACUGAAACACCUUGACA
1884

7437
UUUCUUUACUUCUUUUGAA
1416
7437
UUUCUUUACUUCUUUUGAA
1416
7455
UUCAAAAGAAGUAAAGAAA
1885

7455
AAAUCUGUUUGCAAGGGGA
1417
7455
AAAUCUGUUUGCAAGGGGA
1417
7473
UCCCCUUGCAAACAGAUUU
1886

7473
AAGGACCAUUUCGUAAUGG
1418
7473
AAGGACCAUUUCGUAAUGG
1418
7491
CCAUUACGAAAUGGUCCUU
1887

7491
GUCUGACACAAAAGCAAGU
1419
7491
GUCUGACACAAAAGCAAGU
1419
7509
ACUUGCUUUUGUGUCAGAC
1888

7509
UUUGAUUUUUGCAGCACUA
1420
7509
UUUGAUUUUUGCAGCACUA
1420
7527
UAGUGCUGCAAAAAUCAAA
1889

7527
AGCAAUGGACUUUGUUGUU
1421
7527
AGCAAUGGACUUUGUUGUU
1421
7545
AACAACAAAGUCCAUUGCU
1890

7545
UUUUCUUUUUGAUCAGAAC
1422
7545
UUUUCUUUUUGAUCAGAAC
1422
7563
GUUCUGAUCAAAAAGAAAA
1891

7563
CAUUCCUUCUUUACUGGUC
1423
7563
CAUUCCUUCUUUACUGGUC
1423
7581
GACCAGUAAAGAAGGAAUG
1892

7581
CACAGCCACGUGCUCAUUC
1424
7581
CACAGCCACGUGCUCAUUC
1424
7599
GAAUGAGCACGUGGCUGUG
1893

7599
CCAUUCUUCUUUUUGUAGA
1425
7599
CCAUUCUUCUUUUUGUAGA
1425
7617
UCUACAAAAAGAAGAAUGG
1894

7617
ACUUUGGGCCCACGUGUUU
1426
7617
ACUUUGGGCCCACGUGUUU
1426
7635
AAACACGUGGGCCCAAAGU
1895

7635
UUAUGGGCAUUGAUACAUA
1427
7635
UUAUGGGCAUUGAUACAUA
1427
7653
UAUGUAUCAAUGCCCAUAA
1896

7653
AUAUAAAUAUAUAGAUAUA
1428
7653
AUAUAAAUAUAUAGAUAUA
1428
7671
UAUAUCUAUAUAUUUAUAU
1897

7671
AAAUAUAUAUGAAUAUAUU
1429
7671
AAAUAUAUAUGAAUAUAUU
1429
7689
AAUAUAUUCAUAUAUAUUU
1898

7689
UUUUUUAAGUUUCCUACAC
1430
7689
UUUUUUAAGUUUCCUACAC
1430
7707
GUGUAGGAAACUUAAAAAA
1899

7707
CCUGGAGGUUGCAUGGACU
1431
7707
CCUGGAGGUUGCAUGGACU
1431
7725
AGUCCAUGCAACCUCCAGG
1900

7725
UGUACGACCGGCAUGACUU
1432
7725
UGUACGACCGGCAUGACUU
1432
7743
AAGUCAUGCCGGUCGUACA
1901

7743
UUAUAUUGUAUACAGAUUU
1433
7743
UUAUAUUGUAUACAGAUUU
1433
7761
AAAUCUGUAUACAAUAUAA
1902

7761
UUGCACGCCAAACUCGGCA
1434
7761
UUGCACGCCAAACUCGGCA
1434
7779
UGCCGAGUUUGGCGUGCAA
1903

7779
AGCUUUGGGGAAGAAGAAA
1435
7779
AGCUUUGGGGAAGAAGAAA
1435
7797
UUUCUUCUUCCCCAAAGCU
1904

7797
AAAUGCCUUUCUGUUCCCC
1436
7797
AAAUGCCUUUCUGUUCCCC
1436
7815
GGGGAACAGAAAGGCAUUU
1905

7815
CUCUCAUGACAUUUGCAGA
1437
7815
CUCUCAUGACAUUUGCAGA
1437
7833
UCUGCAAAUGUCAUGAGAG
1906

7833
AUACAAAAGAUGGAAAUUU
1438
7833
AUACAAAAGAUGGAAAUUU
1438
7851
AAAUUUCCAUCUUUUGUAU
1907

7851
UUUCUGUAAAACAAAACCU
1439
7851
UUUCUGUAAAACAAAACCU
1439
7869
AGGUUUUGUUUUACAGAAA
1908

7869
UUGAAGGAGAGGAGGGCGG
1440
7869
UUGAAGGAGAGGAGGGCGG
1440
7887
CCGCCCUCCUCUCCUUCAA
1909

7887
GGGAAGUUUGCGUCUUAUU
1441
7887
GGGAAGUUUGCGUCUUAUU
1441
7905
AAUAAGACGCAAACUUCCC
1910

7905
UGAACUUAUUCUUAAGAAA
1442
7905
UGAACUUAUUCUUAAGAAA
1442
7923
UUUCUUAAGAAUAAGUUCA
1911

7923
AUUGUACUUUUUAUUGUAA
1443
7923
AUUGUACUUUUUAUUGUAA
1443
7941
UUACAAUAAAAAGUACAAU
1912

7941
AGAAAAAUAAAAAGGACUA
1444
7941
AGAAAAAUAAAAAGGACUA
1444
7959
UAGUCCUUUUUAUUUUUCU
1913

7959
ACUUAAACAUUUGUCAUAU
1445
7959
ACUUAAACAUUUGUCAUAU
1445
7977
AUAUGACAAAUGUUUAAGU
1914

7977
UUAAGAAAAAAAGUUUAUC
1446
7977
UUAAGAAAAAAAGUUUAUC
1446
7995
GAUAAACUUUUUUUCUUAA
1915

7995
CUAGCACUUGUGACAUACC
1447
7995
CUAGCACUUGUGACAUACC
1447
8013
GGUAUGUCACAAGUGCUAG
1916

8013
CAAUAAUAGAGUUUAUUGU
1448
8013
CAAUAAUAGAGUUUAUUGU
1448
8031
ACAAUAAACUCUAUUAUUG
1917

8031
UAUUUAUGUGGAAACAGUG
1449
8031
UAUUUAUGUGGAAACAGUG
1449
8049
CACUGUUUCCACAUAAAUA
1918

8049
GUUUUAGGGAAACUACUCA
1450
8049
GUUUUAGGGAAACUACUCA
1450
8067
UGAGUAGUUUCCCUAAAAC
1919

8067
AGAAUUCACAGUGAACUGC
1451
8067
AGAAUUCACAGUGAACUGC
1451
8085
GCAGUUCACUGUGAAUUCU
1920

8085
CCUGUCUCUCUCGAGUUGA
1452
8085
CCUGUCUCUCUCGAGUUGA
1452
8103
UCAACUCGAGAGAGACAGG
1921

8103
AUUUGGAGGAAUUUUGUUU
1453
8103
AUUUGGAGGAAUUUUGUUU
1453
8121
AAACAAAAUUCCUCCAAAU
1922

8121
UUGUUUUGUUUUGUUUGUU
1454
8121
UUGUUUUGUUUUGUUUGUU
1454
8139
AACAAACAAAACAAAACAA
1923

8139
UUCCUUUUAUCUCCUUCCA
1455
8139
UUCCUUUUAUCUCCUUCCA
1455
8157
UGGAAGGAGAUAAAAGGAA
1924

8157
ACGGGCCAGGCGAGCGCCG
1456
8157
ACGGGCCAGGCGAGCGCCG
1456
8175
CGGCGCUCGCCUGGCCCGU
1925

8175
GCCCGCCCUCACUGGCCUU
1457
8175
GCCCGCCCUCACUGGCCUU
1457
8193
AAGGCCAGUGAGGGCGGGC
1926

8193
UGUGACGGUUUAUUCUGAU
1458
8193
UGUGACGGUUUAUUCUGAU
1458
8211
AUCAGAAUAAACCGUCACA
1927

8211
UUGAGAACUGGGCGGACUC
1459
8211
UUGAGAACUGGGCGGACUC
1459
8229
GAGUCCGCCCAGUUCUCAA
1928

8229
CGAAAGAGUCCCCUUUUCC
1460
8229
CGAAAGAGUCCCCUUUUCC
1460
8247
GGAAAAGGGGACUCUUUCG
1929

8247
CGCACAGCUGUGUUGACUU
1461
8247
CGCACAGCUGUGUUGACUU
1461
8265
AAGUCAACACAGCUGUGCG
1930

8265
UUUUAAUUACUUUUAGGUG
1462
8265
UUUUAAUUACUUUUAGGUG
1462
8283
CACCUAAAAGUAAUUAAAA
1931

8283
GAUGUAUGGCUAAGAUUUC
1463
8283
GAUGUAUGGCUAAGAUUUC
1463
8301
GAAAUCUUAGCCAUACAUC
1932

8301
CACUUUAAGCAGUCGUGAA
1464
8301
CACUUUAAGCAGUCGUGAA
1464
8319
UUCACGACUGCUUAAAGUG
1933

8319
ACUGUGCGAGCACUGUGGU
1465
8319
ACUGUGCGAGCACUGUGGU
1465
8337
ACCACAGUGCUCGCACAGU
1934

8337
UUUACAAUUAUACUUUGCA
1466
8337
UUUACAAUUAUACUUUGCA
1466
8355
UGCAAAGUAUAAUUGUAAA
1935

8355
AUCGAAAGGAAACCAUUUC
1467
8355
AUCGAAAGGAAACCAUUUC
1467
8373
GAAAUGGUUUCCUUUCGAU
1936

8373
CUUCAUUGUAACGAAGCUG
1468
8373
CUUCAUUGUAACGAAGCUG
1468
8391
CAGCUUCGUUACAAUGAAG
1937

8391
GAGCGUGUUCUUAGCUCGG
1469
8391
GAGCGUGUUCUUAGCUCGG
1469
8409
CCGAGCUAAGAACACGCUC
1938

8409
GCCUCACUUUGUCUCUGGC
1470
8409
GCCUCACUUUGUCUCUGGC
1470
8427
GCCAGAGACAAAGUGAGGC
1939

8427
CAUUGAUUAAAAGUCUGCU
1471
8427
CAUUGAUUAAAAGUCUGCU
1471
8445
AGCAGACUUUUAAUCAAUG
1940

HDAC5 variant 1: NM_005474.4

3
AAUGUUGUUGUUGGUGGCG
2053
3
AAUGUUGUUGUUGGUGGCG
2053
21
CGCCACCAACAACAACAUU
2348

21
GGCGGCGAGCGGAGCCGGA
2054
21
GGCGGCGAGCGGAGCCGGA
2054
39
UCCGGCUCCGCUCGCCGCC
2349

39
AGGAGCCGCCGCAAAGAUG
2055
39
AGGAGCCGCCGCAAAGAUG
2055
57
CAUCUUUGCGGCGGCUCCU
2350

57
GGAGGAGCCGUCGAGGAGG
2056
57
GGAGGAGCCGUCGAGGAGG
2056
75
CCUCCUCGACGGCUCCUCC
2351

75
GUGCUGCCGCCGCUGCCGC
2057
75
GUGCUGCCGCCGCUGCCGC
2057
93
GCGGCAGCGGCGGCAGCAC
2352

93
CCGCCGCUGCUGCCGCCGC
2058
93
CCGCCGCUGGUGCCGCCGC
2058
111
GGGGCGGCAGCAGCGGCGG
2353

111
GCGCCCGGGAAGCCGGAGC
2059
111
CCGCCCGCGAAGCCGGAGC
2059
129
GCUCCGGCUUCGCGGGCGG
2354

129
CUCGAGCCGCAGCGGGGAU
2060
129
CUCGAGCCGCAGCGGGGAU
2060
147
AUCCCCGCUGCGGCUCGAG
2355

147
UGCCGUUCUGAGUGCCUGA
2061
147
UGCCGUUCUGAGUGCCUGA
2061
165
UCAGGCACUCAGAACGGCA
2356

165
ACUGCCUCGCCCCGAAGGA
2062
165
ACUGCCUCGCCCCGAAGGA
2062
183
UCCUUCGGGGCGAGGCAGU
2357

183
AUGGCCUCGGAUGGGCAUU
2063
183
AUGGCCUCGGAUGGGCAUU
2063
201
AAUGCCCAUCCGAGGCCAU
2358

201
UAGAGGCACGGCGGCCCCG
2064
201
UAGAGGCACGGCGGCCCCG
2064
219
CGGGGCCGCCGUGCCUCUA
2359

219
GGGCUCCCGUCCCGUCCGU
2065
219
GGGCUCCCGUCCCGUCCGU
2065
237
ACGGACGGGACGGGAGCCC
2360

237
UCUGUCUGUUAUCGUCUGU
2066
237
UCUGUCUGUUAUCGUCUGU
2066
255
ACAGACGAUAACAGACAGA
2361

255
UCUCUCUUGACAUCACCGC
2067
255
UCUCUCUUGACAUCACCGC
2067
273
GCGGUGAUGUCAAGAGAGA
2362

273
CAGCUCCACCCCCUCCCGU
2068
273
CAGCUCCACCCCCUCCCGU
2068
291
ACGGGAGGGGGUGGAGCUG
2363

291
UCCCAGCCCCCAACGCCAG
2069
291
UCCCAGCCCCCAACGCCAG
2069
309
CUGGCGUUGGGGGCUGGGA
2364

309
GCUUCCUGCAGGCCCAGAG
2070
309
GCUUCCUGCAGGCCCAGAG
2070
327
CUCUGGGCCUGCAGGAAGC
2365

327
GCCGGCAUGAACUCUCCCA
2071
327
GCCGGCAUGAACUCUCCCA
2071
345
UGGGAGAGUUCAUGCCGGC
2366

345
AACGAGUCGGAUGGGAUGU
2072
345
AACGAGUCGGAUGGGAUGU
2072
363
ACAUCCCAUCCGACUCGUU
2367

363
UCAGGUCGGGAACCAUCCU
2073
363
UCAGGUCGGGAACCAUCCU
2073
381
AGGAUGGUUCCCGACCUGA
2368

381
UUGGAAAUCCUGCCGCGGA
2074
381
UUGGAAAUCCUGCCGCGGA
2074
399
UCCGCGGCAGGAUUUCCAA
2369

399
ACUUCUCUGCACAGCAUCC
2075
399
ACUUCUCUGCACAGGAUCC
2075
417
GGAUGCUGUGCAGAGAAGU
2370

417
CCUGUGACAGUGGAGGUGA
2076
417
CCUGUGACAGUGGAGGUGA
2076
435
UCACCUCCACUGUCACAGG
2371

435
AAGCCGGUGCUGCCAAGAG
2077
435
AAGCCGGUGCUGCCAAGAG
2077
453
CUCUUGGCAGCACCGGCUU
2372

453
GCCAUGCCCAGUUCCAUGG
2078
453
GCCAUGCCCAGUUCCAUGG
2078
471
CCAUGGAACUGGGCAUGGC
2373

471
GGGGGUGGGGGUGGAGGCA
2079
471
GGGGGUGGGGGUGGAGGCA
2079
489
UGCCUCCACCCCCACCCCC
2374

489
AGCCCCAGCCCUGUGGAGC
2080
489
AGCCCCAGCCCUGUGGAGC
2080
507
GCUCCACAGGGCUGGGGCU
2375

507
CUACGGGGGGCUCUGGUGG
2081
507
CUACGGGGGGCUCUGGUGG
2081
525
CCACCAGAGCCCCCCGUAG
2376

525
GGCUCUGUGGACCCCACAC
2082
525
GGCUCUGUGGACCCCACAC
2082
543
GUGUGGGGUCCACAGAGCC
2377

543
CUGCGGGAGCAGCAACUGC
2083
543
CUGCGGGAGCAGCAACUGC
2083
561
GCAGUUGCUGCUCCCGCAG
2378

561
CAGCAGGAGCUCCUGGCGC
2084
561
CAGCAGGAGCUCCUGGCGC
2084
579
GCGCCAGGAGCUCCUGCUG
2379

579
CUCAAGCAGCAGCAGCAGC
2085
579
CUCAAGCAGCAGCAGCAGC
2085
597
GCUGCUGCUGCUGCUUGAG
2380

597
CUGCAGAAGCAGCUCCUGU
2086
597
CUGCAGAAGCAGCUCCUGU
2086
615
ACAGGAGCUGCUUCUGCAG
2381

615
UUCGCUGAGUUCCAGAAAC
2087
615
UUCGCUGAGUUCCAGAAAC
2087
633
GUUUCUGGAACUCAGCGAA
2382

633
CAGCAUGACCACCUGACAA
2088
633
CAGCAUGACCACCUGACAA
2088
651
UUGUCAGGUGGUCAUGCUG
2383

651
AGGCAGCAUGAGGUCCAGC
2089
651
AGGCAGCAUGAGGUCCAGC
2089
669
GCUGGACCUCAUGCUGCCU
2384

669
CUGCAGAAGCACCUCAAGC
2090
669
CUGCAGAAGCACCUCAAGC
2090
687
GCUUGAGGUGCUUCUGCAG
2385

687
CAGCAGCAGGAGAUGCUGG
2091
687
CAGCAGCAGGAGAUGCUGG
2091
705
CCAGCAUCUCCUGCUGCUG
2386

705
GCAGCCAAGCAGCAGCAGG
2092
705
GCAGCCAAGCAGCAGCAGG
2092
723
CCUGCUGCUGCUUGGCUGC
2387

723
GAGAUGCUGGCAGCCAAGC
2093
723
GAGAUGCUGGCAGCCAAGC
2093
741
GCUUGGCUGCCAGCAUCUC
2388

741
CGGCAGCAGGAGCUGGAGC
2094
741
CGGCAGCAGGAGCUGGAGC
2094
759
GCUCCAGCUCCUGCUGCCG
2389

759
CAGCAGCGGCAGCGGGAGC
2095
759
CAGCAGCGGCAGCGGGAGC
2095
777
GCUCCCGCUGCCGCUGCUG
2390

777
CAGCAGCGGCAGGAAGAGC
2096
777
CAGCAGCGGCAGGAAGAGC
2096
795
GCUCUUCCUGCCGCUGCUG
2391

795
CUGGAGAAGCAGCGGCUGG
2097
795
CUGGAGAAGCAGCGGCUGG
2097
813
CCAGCCGCUGCUUCUCCAG
2392

813
GAGCAGCAGCUGCUCAUCC
2098
813
GAGCAGCAGCUGCUCAUCC
2098
831
GGAUGAGCAGCUGCUGCUC
2393

831
CUGCGGAACAAGGAGAAGA
2099
831
CUGCGGAACAAGGAGAAGA
2099
849
UCUUCUCCUUGUUCCGCAG
2394

849
AGCAAAGAGAGUGCCAUUG
2100
849
AGCAAAGAGAGUGCCAUUG
2100
867
CAAUGGCACUCUCUUUGCU
2395

867
GCCAGCACUGAGGUAAAGC
2101
867
GCCAGCACUGAGGUAAAGC
2101
885
GCUUUACCUCAGUGCUGGC
2396

885
CUGAGGCUCCAGGAAUUCC
2102
885
CUGAGGCUCCAGGAAUUCC
2102
903
GGAAUUCCUGGAGCCUCAG
2397

903
CUCUUGUCGAAGUCAAAGG
2103
903
CUCUUGUCGAAGUCAAAGG
2103
921
CCUUUGACUUCGACAAGAG
2398

921
GAGCCCACACCAGGCGGCC
2104
921
GAGCCCACACCAGGCGGCC
2104
939
GGCCGCCUGGUGUGGGCUC
2399

939
CUCAACCAUUCCCUCCCAC
2105
939
CUCAACCAUUCCCUCCCAC
2105
957
GUGGGAGGGAAUGGUUGAG
2400

957
CAGCACCCCAAAUGCUGGG
2106
957
CAGCACCCCAAAUGCUGGG
2106
975
CCCAGCAUUUGGGGUGCUG
2401

975
GGAGCCCACCAUGCUUCUU
2107
975
GGAGCCCACCAUGCUUCUU
2107
993
AAGAAGCAUGGUGGGCUCC
2402

993
UUGGACCAGAGUUCCCCUC
2108
993
UUGGACCAGAGUUCCCCUC
2108
1011
GAGGGGAACUCUGGUCCAA
2403

1011
CCCCAGAGCGGCCCCCCUG
2109
1011
CCCCAGAGCGGCCCCCCUG
2109
1029
CAGGGGGGCCGCUCUGGGG
2404

1029
GGGACGCCUCCGUCCUACA
2110
1029
GGGACGCCUCCCUCCUACA
2110
1047
UGUAGGAGGGAGGCGUCCC
2405

1047
AAACUGCCUUUGCCUGGGC
2111
1047
AAACUGCCUUUGCCUGGGC
2111
1065
GCCCAGGCAAAGGCAGUUU
2406

1065
CCGUACGACAGUCGAGACG
2112
1065
CCCUACGACAGUCGAGACG
2112
1083
CGUGUCGACUGUCGUAGGG
2407

1083
GACUUCCCCCUCCGCAAAA
2113
1083
GACUUCCCCCUCCGCAAAA
2113
1101
UUUUGCGGAGGGGGAAGUC
2408

1101
ACAGCCUCUGAACCCAACU
2114
1101
ACAGCCUCUGAACCCAACU
2114
1119
AGUUGGGUUCAGAGGCUGU
2409

1119
UUGAAAGUGCGUUCAAGGC
2115
1119
UUGAAAGUGCGUUCAAGGC
2115
1137
GCCUUGAACGCACUUUCAA
2410

1137
CUAAAACAGAAGGUGGCUG
2116
1137
CUAAAACAGAAGGUGGCUG
2116
1155
CAGCCACCUUCUGUUUUAG
2411

1155
GAGCGGAGAAGCAGUCCCC
2117
1155
GAGCGGAGAAGCAGUCCCC
2117
1173
GGGGACUGCUUCUCCGCUC
2412

1173
CUCCUGCGUCGCAAGGAUG
2118
1173
CUCCUGCGUCGCAAGGAUG
2118
1191
CAUCCUUGCGACGCAGGAG
2413

1191
GGGACUGUUAUUAGCACCU
2119
1191
GGGACUGUUAUUAGCACCU
2119
1209
AGGUGCUAAUAACAGUCCC
2414

1209
UUUAAGAAGAGAGCUGUUG
2120
1209
UUUAAGAAGAGAGCUGUUG
2120
1227
CAACAGCUCUCUUCUUAAA
2415

1227
GAGAUCACAGGUGCCGGGC
2121
1227
GAGAUCACAGGUGCCGGGC
2121
1245
GCCCGGCACCUGUGAUCUC
2416

1245
CCUGGGGCGUCGUCCGUGU
2122
1245
CCUGGGGCGUCGUCCGUGU
2122
1263
ACACGGACGACGCCCCAGG
2417

1263
UGUAACAGCGCACCCGGCU
2123
1263
UGUAACAGCGCACCCGGCU
2123
1281
AGCCGGGUGCGCUGUUACA
2418

1281
UCCGGCCCCAGCUCUCCCA
2124
1281
UCCGGCCCCAGCUCUCCCA
2124
1299
UGGGAGAGCUGGGGCCGGA
2419

1299
AACAGCUCCCACAGCACCA
2125
1299
AACAGCUCCCACAGCACCA
2125
1317
UGGUGCUGUGGGAGCUGUU
2420

1317
AUCGCUGAGAAUGGCUUUA
2126
1317
AUCGCUGAGAAUGGCUUUA
2126
1335
UAAAGCCAUUCUCAGCGAU
2421

1335
ACUGGCUCAGUCCCCAACA
2127
1335
ACUGGCUCAGUCCCCAACA
2127
1353
UGUUGGGGACUGAGCCAGU
2422

1353
AUCCCCACUGAGAUGCUCC
2128
1353
AUCCCCACUGAGAUGCUCC
2128
1371
GGAGCAUCUCAGUGGGGAU
2423

1371
CCUCAGCACCGAGCCCUCC
2129
1371
CCUCAGCACCGAGCCCUCC
2129
1389
GGAGGGCUCGGUGCUGAGG
2424

1389
CCUCUGGACAGCUCCCCCA
2130
1389
CCUCUGGACAGCUCCCCCA
2130
1407
UGGGGGAGCUGUCCAGAGG
2425

1407
AACCAGUUCAGCCUCUACA
2131
1407
AACCAGUUCAGCCUCUACA
2131
1425
UGUAGAGGCUGAACUGGUU
2426

1425
ACGUCUCCUUCUCUGCCCA
2132
1425
ACGUCUCCUUCUCUGCCCA
2132
1443
UGGGCAGAGAAGGAGACGU
2427

1443
AACAUCUCCCUAGGGCUGC
2133
1443
AACAUCUCCCUAGGGCUGC
2133
1461
GCAGCCCUAGGGAGAUGUU
2428

1461
CAGGCCACGGUCACUGUCA
2134
1461
CAGGCCACGGUCACUGUCA
2134
1479
UGACAGUGACCGUGGCCUG
2429

1479
ACCAACUCACACCUCACUG
2135
1479
ACCAACUCACACCUCACUG
2135
1497
CAGUGAGGUGUGAGUUGGU
2430

1497
GCCUCCCCGAAGCUGUCGA
2136
1497
GCCUCCCCGAAGCUGUCGA
2136
1515
UCGACAGCUUCGGGGAGGC
2431

1515
ACACAGCAGGAGGCCGAGA
2137
1515
ACACAGCAGGAGGCCGAGA
2137
1533
UCUCGGCCUCCUGCUGUGU
2432

1533
AGGCAGGCCCUCCAGUCCC
2138
1533
AGGCAGGCCCUCCAGUCCC
2138
1551
GGGACUGGAGGGCCUGCCU
2433

1551
CUGCGGCAGGGUGGCACGC
2139
1551
CUGCGGCAGGGUGGCACGC
2139
1569
GCGUGCCACCCUGCCGCAG
2434

1569
CUGACCGGCAAGUUCAUGA
2140
1569
CUGACCGGCAAGUUCAUGA
2140
1587
UCAUGAACUUGCCGGUCAG
2435

1587
AGCACAUCCUCUAUUCCUG
2141
1587
AGCACAUCCUCUAUUCCUG
2141
1605
CAGGAAUAGAGGAUGUGCU
2436

1605
GGCUGCCUGCUGGGCGUGG
2142
1605
GGCUGCCUGCUGGGCGUGG
2142
1623
CCACGCCCAGCAGGCAGCC
2437

1623
GCACUGGAGGGCGACGGGA
2143
1623
GCACUGGAGGGCGACGGGA
2143
1641
UCCCGUCGCCCUCCAGUGC
2438

1641
AGCCCCCACGGGCAUGCCU
2144
1641
AGCCCCCACGGGCAUGCCU
2144
1659
AGGCAUGCCCGUGGGGGCU
2439

1659
UCCCUGCUGCAGCAUGUGC
2145
1659
UCCCUGCUGCAGCAUGUGC
2145
1677
GCACAUGCUGCAGCAGGGA
2440

1677
CUGUUGCUGGAGCAGGCCC
2146
1677
CUGUUGCUGGAGCAGGCCC
2146
1695
GGGCCUGCUCCAGCAACAG
2441

1695
CGGCAGCAGAGCACCCUCA
2147
1695
CGGCAGCAGAGCACCCUCA
2147
1713
UGAGGGUGCUCUGCUGCCG
2442

1713
AUUGCUGUGCCACUCCACG
2148
1713
AUUGCUGUGCCACUCCACG
2148
1731
CGUGGAGUGGCACAGCAAU
2443

1731
GGGCAGUCCCCACUAGUGA
2149
1731
GGGCAGUCCCCACUAGUGA
2149
1749
UCACUAGUGGGGACUGCCC
2444

1749
ACGGGUGAACGUGUGGCCA
2150
1749
ACGGGUGAACGUGUGGCCA
2150
1767
UGGCCACACGUUCACCCGU
2445

1767
ACCAGCAUGCGGACGGUAG
2151
1767
ACCAGCAUGCGGACGGUAG
2151
1785
CUACCGUCCGCAUGCUGGU
2446

1785
GGCAAGCUCCCGCGGCAUC
2152
1785
GGCAAGCUCCCGCGGCAUC
2152
1803
GAUGCCGCGGGAGCUUGCC
2447

1803
CGGCCCCUGAGCCGCACUC
2153
1803
CGGCCCCUGAGCCGCACUC
2153
1821
GAGUGCGGCUCAGGGGCCG
2448

1821
CAGUCCUCACCGCUGCCGC
2154
1821
CAGUCCUCACCGCUGCCGC
2154
1839
GCGGCAGCGGUGAGGACUG
2449

1839
CAGAGUCCCCAGGCCCUGC
2155
1839
CAGAGUCCCCAGGCCCUGC
2155
1857
GCAGGGCCUGGGGACUCUG
2450

1857
CAGCAGCUGGUCAUGCAAC
2156
1857
CAGCAGCUGGUCAUGCAAC
2156
1875
GUUGCAUGACCAGCUGCUG
2451

1875
CAACAGCACCAGCAGUUCC
2157
1875
CAACAGCACCAGCAGUUCC
2157
1893
GGAACUGCUGGUGCUGUUG
2452

1893
CUGGAGAAGCAGAAGCAGC
2158
1893
CUGGAGAAGCAGAAGCAGC
2158
1911
GCUGCUUCUGCUUCUCCAG
2453

1911
CAGCAGCUACAGCUGGGCA
2159
1911
CAGCAGCUACAGCUGGGCA
2159
1929
UGCCCAGCUGUAGCUGCUG
2454

1929
AAGAUCCUCACCAAGACAG
2160
1929
AAGAUCCUCACCAAGACAG
2160
1947
CUGUCUUGGUGAGGAUCUU
2455

1947
GGGGAGCUGCCCAGGCAGC
2161
1947
GGGGAGCUGCCCAGGCAGC
2161
1965
GCUGCCUGGGCAGCUCCCC
2456

1965
CCCACCACCCACCCUGAGG
2162
1965
CCCACCACCCACCCUGAGG
2162
1983
CCUCAGGGUGGGUGGUGGG
2457

1983
GAGACAGAGGAGGAGGUGA
2163
1983
GAGACAGAGGAGGAGCUGA
2163
2001
UCAGCUCCUCCUCUGUCUC
2458

2001
ACGGAGCAGCAGGAGGUCU
2164
2001
ACGGAGCAGCAGGAGGUCU
2164
2019
AGACCUCCUGCUGCUCCGU
2459

2019
UUGCUGGGGGAGGGAGCCC
2165
2019
UUGCUGGGGGAGGGAGCCC
2165
2037
GGGCUCCCUCCCCCAGCAA
2460

2037
CUGACCAUGCCCCGGGAGG
2166
2037
CUGACCAUGCCCCGGGAGG
2166
2055
CCUCCCGGGGCAUGGUCAG
2461

2055
GGCUCCACAGAGAGUGAGA
2167
2055
GGCUCCACAGAGAGUGAGA
2167
2073
UCUCACUCUCUGUGGAGCC
2462

2073
AGCACACAGGAAGACCUGG
2168
2073
AGCACACAGGAAGACCUGG
2168
2091
CCAGGUCUUCCUGUGUGCU
2463

2091
GAGGAGGAGGACGAGGAAG
2169
2091
GAGGAGGAGGACGAGGAAG
2169
2109
CUUCCUCGUCCUCCUCCUC
2464

2109
GACGAUGGGGAGGAGGAGG
2170
2109
GACGAUGGGGAGGAGGAGG
2170
2127
CCUCCUCCUCCCCAUCGUC
2465

2127
GAGGAUUGCAUCCAGGUUA
2171
2127
GAGGAUUGCAUCCAGGUUA
2171
2145
UAACCUGGAUGCAAUCCUC
2466

2145
AAGGACGAGGAGGGCGAGA
2172
2145
AAGGACGAGGAGGGGGAGA
2172
2163
UCUCGCCCUCCUCGUCCUU
2467

2163
AGUGGUGCUGAGGAGGGGC
2173
2163
AGUGGUGCUGAGGAGGGGC
2173
2181
GCCCCUCCUCAGCACCACU
2468

2181
CCCGACUUGGAGGAGCCUG
2174
2181
GCCGACUUGGAGGAGCCUG
2174
2199
CAGGCUCCUCCAAGUCGGG
2469

2199
GGUGCUGGAUACAAAAAAC
2175
2199
GGUGCUGGAUACAAAAAAC
2175
2217
GUUUUUUGUAUCCAGCACC
2470

2217
CUGUUCUCAGAUGCCCAGC
2176
2217
CUGUUCUCAGAUGCCCAGC
2176
2235
GCUGGGCAUCUGAGAACAG
2471

2235
CCGCUGCAGCCUUUGCAGG
2177
2235
CCGCUGCAGCCUUUGCAGG
2177
2253
CCUGCAAAGGCUGCAGCGG
2472

2253
GUGUACCAGGCGCCCCUCA
2178
2253
GUGUACCAGGCGCCCCUCA
2178
2271
UGAGGGGCGCCUGGUACAC
2473

2271
AGCCUGGCCACUGUGCCCC
2179
2271
AGCCUGGCCACUGUGCCCC
2179
2289
GGGGCACAGUGGCCAGGCU
2474

2289
CACCAGGCCCUGGGCCGUA
2180
2289
CACCAGGCCCUGGGCCGUA
2180
2307
UACGGCCCAGGGCCUGGUG
2475

2307
ACCCAGUCCUCCCCUGCUG
2181
2307
ACCCAGUCCUCCCCUGCUG
2181
2325
CAGCAGGGGAGGACUGGGU
2476

2325
GCCCCUGGGGGCAUGAAGA
2182
2325
GCCCCUGGGGGCAUGAAGA
2182
2343
UCUUCAUGCCCCCAGGGGC
2477

2343
AGCCCCCCAGACCAGCCCG
2183
2343
AGCCCCCCAGACCAGCCCG
2183
2361
CGGGCUGGUCUGGGGGGCU
2478

2361
GUCAAGCACCUCUUCACCA
2184
2361
GUCAAGCACCUCUUCACCA
2184
2379
UGGUGAAGAGGUGCUUGAC
2479

2379
ACAGGUGUGGUCUAGGACA
2185
2379
ACAGGUGUGGUCUACGACA
2185
2397
UGUCGUAGACCACACCUGU
2480

2397
ACGUUCAUGCUAAAGCACC
2186
2397
ACGUUCAUGCUAAAGCACC
2186
2415
GGUGCUUUAGCAUGAACGU
2481

2415
CAGUGCAUGUGCGGGAACA
2187
2415
CAGUGCAUGUGCGGGAACA
2187
2433
UGUUCCCGCACAUGCACUG
2482

2433
ACACACGUGCACCCUGAGC
2188
2433
ACACACGUGCACCCUGAGC
2188
2451
GCUCAGGGUGCACGUGUGU
2483

2451
CAUGCUGGCCGGAUCCAGA
2189
2451
CAUGCUGGCCGGAUCCAGA
2189
2469
UCUGGAUCCGGCCAGCAUG
2484

2469
AGCAUCUGGUCCCGGCUGC
2190
2469
AGCAUCUGGUCCCGGCUGC
2190
2487
GCAGCCGGGACCAGAUGCU
2485

2487
CAGGAGACAGGCCUGCUUA
2191
2487
CAGGAGACAGGCCUGCUUA
2191
2505
UAAGCAGGCCUGUCUCCUG
2486

2505
AGCAAGUGCGAGCGGAUCC
2192
2505
AGCAAGUGCGAGCGGAUCC
2192
2523
GGAUCCGCUCGCACUUGCU
2487

2523
CGAGGUCGCAAAGCCACGC
2193
2523
CGAGGUCGCAAAGCCACGC
2193
2541
GCGUGGCUUUGCGACCUCG
2488

2541
CUAGAUGAGAUCCAGACAG
2194
2541
CUAGAUGAGAUCCAGACAG
2194
2559
CUGUCUGGAUCUCAUCUAG
2489

2559
GUGCACUCUGAAUACCACA
2195
2559
GUGCACUCUGAAUACCACA
2195
2577
UGUGGUAUUCAGAGUGCAC
2490

2577
ACCCUGCUCUAUGGGACCA
2196
2577
ACCCUGCUCUAUGGGACCA
2196
2595
UGGUCCCAUAGAGCAGGGU
2491

2595
AGUCCCCUCAACCGGCAGA
2197
2595
AGUCCCCUCAACCGGCAGA
2197
2613
UCUGCCGGUUGAGGGGACU
2492

2613
AAGCUAGACAGCAAGAAGU
2198
2613
AAGCUAGACAGCAAGAAGU
2198
2631
ACUUCUUGCUGUCUAGCUU
2493

2631
UUGCUCGGCCCCAUCAGCC
2199
2631
UUGCUCGGCCCCAUCAGCC
2199
2649
GGCUGAUGGGGCCGAGCAA
2494

2649
CAGAAGAUGUAUGCUGUGC
2200
2649
CAGAAGAUGUAUGCUGUGC
2200
2667
GCACAGCAUACAUCUUCUG
2495

2667
CUGCCUUGUGGGGGCAUCG
2201
2667
CUGCCUUGUGGGGGCAUCG
2201
2685
CGAUGCCCCCACAAGGCAG
2496

2685
GGGGUGGACAGUGACACCG
2202
2685
GGGGUGGACAGUGACACCG
2202
2703
CGGUGUCACUGUCCACCCC
2497

2703
GUGUGGAAUGAGAUGCACU
2203
2703
GUGUGGAAUGAGAUGCACU
2203
2721
AGUGCAUCUCAUUCCACAC
2498

2721
UCCUCCAGUGCUGUGCGCA
2204
2721
UCCUCCAGUGCUGUGCGCA
2204
2739
UGCGCACAGCACUGGAGGA
2499

2739
AUGGCAGUGGGCUGCCUGC
2205
2739
AUGGCAGUGGGCUGCCUGC
2205
2757
GCAGGCAGCCCACUGCCAU
2500

2757
CUGGAGCUGGCCUUCAAGG
2206
2757
CUGGAGCUGGGCUUCAAGG
2206
2775
CCUUGAAGGCCAGGUCCAG
2501

2775
GUGGCUGCAGGAGAGCUCA
2207
2775
GUGGCUGCAGGAGAGCUCA
2207
2793
UGAGCUCUCCUGCAGCCAC
2502

2793
AAGAAUGGAUUUGCCAUCA
2208
2793
AAGAAUGGAUUUGCCAUCA
2208
2811
UGAUGGCAAAUCCAUUCUU
2503

2811
AUCCGGCCCCCAGGACACC
2209
2811
AUCCGGCCCCCAGGACACC
2209
2829
GGUGUCCUGGGGGCCGGAU
2504

2829
CACGCCGAGGAAUCCACAG
2210
2829
CACGCCGAGGAAUCCACAG
2210
2847
CUGUGGAUUCCUCGGCGUG
2505

2847
GCCAUGGGAUUCUGCUUCU
2211
2847
GCCAUGGGAUUCUGCUUCU
2211
2865
AGAAGCAGAAUCCCAUGGC
2506

2865
UUCAACUCUGUAGCCAUCA
2212
2865
UUCAACUCUGUAGCCAUCA
2212
2883
UGAUGGCUACAGAGUUGAA
2507

2883
ACCGCAAAACUCCUACAGC
2213
2883
ACCGCAAAACUCCUACAGC
2213
2901
GCUGUAGGAGUUUUGCGGU
2508

2901
CAGAAGUUGAACGUGGGCA
2214
2901
CAGAAGUUGAACGUGGGCA
2214
2919
UGCCCACGUUCAACUUCUG
2509

2919
AAGGUCCUCAUCGUGGACU
2215
2919
AAGGUCCUCAUCGUGGACU
2215
2937
AGUCCACGAUGAGGACCUU
2510

2937
UGGGACAUUCACCAUGGCA
2216
2937
UGGGACAUUCACCAUGGCA
2216
2955
UGCCAUGGUGAAUGUCCCA
2511

2955
AAUGGCACCCAGCAGGCGU
2217
2955
AAUGGCACCCAGCAGGCGU
2217
2973
ACGCCUGCUGGGUGGCAUU
2512

2973
UUCUACAAUGACCCCUCUG
2218
2973
UUCUACAAUGACCCCUCUG
2218
2991
CAGAGGGGUCAUUGUAGAA
2513

2991
GUGCUCUACAUCUCUCUGC
2219
2991
GUGCUCUACAUCUCUCUGC
2219
3009
GCAGAGAGAUGUAGAGGAC
2514

3009
CAUCGCUAUGACAACGGGA
2220
3009
CAUCGCUAUGACAACGGGA
2220
3027
UCCCGUUGUCAUAGCGAUG
2515

3027
AACUUCUUUCCAGGCUCUG
2221
3027
AACUUCUUUCCAGGCUCUG
2221
3045
CAGAGCCUGGAAAGAAGUU
2516

3045
GGGGCUCCUGAAGAGGUUG
2222
3045
GGGGCUCCUGAAGAGGUUG
2222
3063
CAACCUCUUCAGGAGCCCC
2517

3063
GGUGGAGGACCAGGCGUGG
2223
3063
GGUGGAGGACCAGGCGUGG
2223
3081
CCACGCCUGGUCCUCCACC
2518

3081
GGGUACAAUGUGAACGUGG
2224
3081
GGGUACAAUGUGAACGUGG
2224
3099
CCACGUUCACAUUGUACCC
2519

3099
GCAUGGACAGGAGGUGUGG
2225
3099
GCAUGGACAGGAGGUGUGG
2225
3117
CCACACCUCCUGUCCAUGC
2520

3117
GACCCCCCCAUUGGAGACG
2226
3117
GACCCCCCCAUUGGAGACG
2226
3135
CGUCUCCAAUGGGGGGGUC
2521

3135
GUGGAGUACCUUACAGCCU
2227
3135
GUGGAGUACCUUACAGCCU
2227
3153
AGGCUGUAAGGUACUCCAC
2522

3153
UUCAGGACAGUGGUGAUGC
2228
3153
UUCAGGACAGUGGUGAUGC
2228
3171
GCAUCACCACUGUCCUGAA
2523

3171
CCCAUUGCCCACGAGUUCU
2229
3171
CCCAUUGCCCACGAGUUCU
2229
3189
AGAACUCGUGGGCAAUGGG
2524

3189
UCACCUGAUGUGGUCCUAG
2230
3189
UCACCUGAUGUGGUCCUAG
2230
3207
CUAGGACCACAUCAGGUGA
2525

3207
GUCUCCGCCGGGUUUGAUG
2231
3207
GUCUCCGCCGGGUUUGAUG
2231
3225
CAUCAAACCCGGCGGAGAC
2526

3225
GCUGUUGAAGGACAUCUGU
2232
3225
GCUGUUGAAGGACAUCUGU
2232
3243
ACAGAUGUCCUUCAACAGC
2527

3243
UCUCCUCUGGGUGGCUACU
2233
3243
UCUCCUCUGGGUGGCUACU
2233
3261
AGUAGCCACCCAGAGGAGA
2528

3261
UCUGUCACCGCCAGAUGUU
2234
3261
UCUGUCACCGCCAGAUGUU
2234
3279
AACAUCUGGCGGUGACAGA
2529

3279
UUUGGCCACUUGACCAGGC
2235
3279
UUUGGCCACUUGACCAGGC
2235
3297
GCCUGGUCAAGUGGCCAAA
2530

3297
CAGCUGAUGACCCUGGCAG
2236
3297
CAGCUGAUGACCCUGGCAG
2236
3315
CUGCCAGGGUCAUCAGCUG
2531

3315
GGGGGCCGGGUGGUGCUGG
2237
3315
GGGGGCCGGGUGGUGCUGG
2237
3333
CCAGCACCACCCGGCCCCC
2532

3333
GCCCUGGAGGGAGGCCAUG
2238
3333
GCCCUGGAGGGAGGCCAUG
2238
3351
CAUGGCCUCCCUCCAGGGC
2533

3351
GACUUGACCGCCAUCUGUG
2239
3351
GACUUGACCGCCAUCUGUG
2239
3369
CACAGAUGGCGGUCkAGUC
2534

3369
GAUGCCUCUGAGGCUUGUG
2240
3369
GAUGCCUCUGAGGCUUGUG
2240
3387
CACAAGCCUCAGAGGCAUC
2535

3387
GUCUCGGCUCUGCUCAGUG
2241
3387
GUCUCGGCUCUGCUCAGUG
2241
3405
CACUGAGCAGAGCCGAGAC
2536

3405
GAUGAGCUGCAGCCCUUGG
2242
3405
GAUGAGCUGCAGCCCUUGG
2242
3423
CCAAGGGCUGCAGCUCUAC
2537

3423
GAUGAGGCAGUCUUGCAGC
2243
3423
GAUGAGGCAGUCUUGCAGC
2243
3441
GCUGCAAGACUGCCUCAUC
2538

3441
CAAAAGCCCAACAUCAACG
2244
3441
CAAAAGCCCAACAUCAACG
2244
3459
CGUUGAUGUUGGGCUUUUG
2539

3459
GCAGUGGCCACGCUAGAGA
2245
3459
GCAGUGGCCACGCUAGAGA
2245
3477
UCUCUAGCGUGGCCACUGC
2540

3477
AAAGUCAUCGAGAUCCAGA
2246
3477
AAAGUCAUCGAGAUCCAGA
2246
3495
UCUGGAUCUCGAUGACUUU
2541

3495
AGCAAACACUGGAGCUGUG
2247
3495
AGCAAACACUGGAGCUGUG
2247
3513
CACAGCUCCAGUGUUUGCU
2542

3513
GUGCAGAAGUUCGCCGCUG
2248
3513
GUGCAGAAGUUCGCCGCUG
2248
3531
CAGCGGCGAACUUCUGCAC
2543

3531
GGUCUGGGCCGGUCCCUGC
2249
3531
GGUCUGGGCCGGUCCCUGC
2249
3549
GCAGGGACCGGCCCAGACC
2544

3549
CGAGAGGCCCAAGCAGGUG
2250
3549
CGAGAGGCCCAAGCAGGUG
2250
3567
CACCUGCUUGGGCCUCUCG
2545

3567
GAGACCGAGGAGGCCGAGA
2251
3567
GAGACCGAGGAGGCCGAGA
2251
3585
UCUCGGCCUCCUCGGUCUC
2546

3585
ACUGUGAGCGCGAUGGCCU
2252
3585
ACUGUGAGCGCCAUGGCCU
2252
3603
AGGCCAUGGCGCUCACAGU
2547

3603
UUGCUGUCGGUGGGGGCCG
2253
3603
UUGCUGUCGGUGGGGGCCG
2253
3621
CGGCCCCCACCGACAGCAA
2548

3621
GAGCAGGCCCAGGCUGCGG
2254
3621
GAGCAGGCCCAGGCUGCGG
2254
3639
CCGCAGCCUGGGCCUGCUC
2549

3639
GCAGCCCGGGAACACAGCC
2255
3639
GCAGCCCGGGAACACAGCC
2255
3657
GGCUGUGUUCCCGGGCUGC
2550

3657
CCCAGGCCGGCAGAGGAGC
2256
3657
CCCAGGCCGGCAGAGGAGC
2256
3675
GCUCCUCUGCCGGCCUGGG
2551

3675
CCCAUGGAGCAGGAGCCUG
2257
3675
CCCAUGGAGCAGGAGCCUG
2257
3693
CAGGCUCCUGCUCCAUGGG
2552

3693
GCCCUGUGACGCCCCGGCC
2258
3693
GCCCUGUGACGCCCCGGCC
2258
3711
GGCCGGGGCGUCACAGGGC
2553

3711
CCCCAUCCCUCUGGGCUUC
2259
3711
CCCCAUCCCUCUGGGCUUC
2259
3729
GAAGCCCAGAGGGAUGGGG
2554

3729
CACCAUUGUGAUUUUGUUU
2260
3729
CACCAUUGAGAUUUUGUUU
2260
3747
AAACAAAAUCACAAUGGUG
2555

3747
UAUUUUUUCUAUUAAAAAC
2261
3747
UAUUUUUUCUAUUAAAAAC
2261
3765
GUUUUUAAUAGAAAAAAUA
2556

3765
CAAAAAGUCACACAUUCAA
2262
3765
CAAAAAGUCACACAUUCAA
2262
3783
UUGAAUGUGUGACUUUUUG
2557

3783
ACAAGGUGUGCCGUGUGGG
2263
3783
ACAAGGUGUGCCGUGUGGG
2263
3801
CCCACACGGCACACCUUGU
2558

3801
GUCUCUCAGCCUUGCCCCU
2264
3801
GUCUCUCAGCCUUGCCCCU
2264
3819
AGGGGCAAGGCUGAGAGAC
2559

3819
UCCUGCUCCUCUACGCUGC
2265
3819
UCCUGCUCCUCUACGCUGC
2265
3837
GCAGCGUAGAGGAGCAGGA
2560

3837
CCUCAGGCCCCCAGCCCUG
2266
3837
CCUCAGGCCCCCAGCCCUG
2266
3855
CAGGGCUGGGGGCCUGAGG
2561

3855
GUGGCUUCCACCUCAGCUC
2267
3855
GUGGCUUCCACCUCAGCUC
2267
3873
GAGCUGAGGUGGAAGCCAC
2562

3873
CUAGAAGCCUGCUCCCUCU
2268
3873
CUAGAAGCCUGCUCCCUCU
2268
3891
AGAGGGAGCAGGCUUCUAG
2563

3891
UGCAGGGGGUGGUGGUGUC
2269
3891
UGCAGGGGGUGGUGGUGUC
2269
3909
GACACCACCACCCCCUGCA
2564

3909
CUUCCCAGCCCUGUCCCAU
2270
3909
CUUCCCAGCCCUGUCCCAU
2270
3927
AUGGGACAGGGCUGGGAAG
2565

3927
UGUGUCCCUCCCCCCAUUU
2271
3927
UGUGUCCCUCCCCCCAUUU
2271
3945
AAAUGGGGGGAGGGACACA
2566

3945
UUCCUGCAUUCUGUCUGUC
2272
3945
UUCCUGCAUUCUGUCUGUC
2272
3963
GACAGACAGAAUGCAGGAA
2567

3963
CCUUUUCCUCCUUGGAGCC
2273
3963
CCUUUUCCUCCUUGGAGCC
2273
3981
GGCUCCAAGGAGGAAAAGG
2568

3981
CUGGGCCAGGUCAAGGUGG
2274
3981
CUGGGCCAGCUCAAGGUGG
2274
3999
CCACCUUGAGCUGGCCCAG
2569

3999
GGCACGGGGGCCCAGACAG
2275
3999
GGCACGGGGGCCCAGACAG
2275
4017
CUGUCUGGGCCCCCGUGCC
2570

4017
GUACUCUCCAGUUCUGGGG
2276
4017
GUACUCUCCAGUUCUGGGG
2276
4035
CCCCAGAACUGGAGAGUAC
2571

4035
GCCCCCCGAGUGAGGAGGG
2277
4035
GCCCCCCGAGUGAGGAGGG
2277
4053
CCCUCCUCACUCGGGGGGC
2572

4053
GAACGGGAAGUCGGUGCCU
2278
4053
GAACGGGAAGUCGGUGCCU
2278
4071
AGGCACCGACUUCCCGUUC
2573

4071
UUGGUUUCAGCUGAUUUGG
2279
4071
UUGGUUUCAGCUGAUUUGG
2279
4089
CCAAAUCAGCUGXAACCAA
2574

4089
GGGGGAAAUGCCUUAAUUU
2280
4089
GGGGGAAAUGCCUUAAUUU
2280
4107
AAAUUAAGGCAUUUCCCCC
2575

4107
UCACUCUCCUCCCUUCUCC
2281
4107
UCACUCUCCUCCCUUCUCC
2281
4125
GGAGAAGGGAGGAGAGUGA
2576

4125
CAGCCUCAGGGGAGGAUCU
2282
4125
CAGCCUCAGGGGAGGAUCU
2282
4143
AGAUCCUCCCCUGAGGCUG
2577

4143
UGGAGGAUCCACUACUGUC
2283
4143
UGGAGGAUCCACUACUGUC
2283
4161
GACAGUAGUGGAUCCUCCA
2578

4161
CUUUAAGAUGCAGAGUGGA
2284
4161
CUUUAAGAUGCAGAGUGGA
2284
4179
UCCACUCUGCAUCUUAAAG
2579

4179
AGGGGAGGUGGGCACCCAC
2285
4179
AGGGGAGGUGGGCACCCAC
2285
4197
GUGGGUGCCCACCUCCCCU
2580

4197
CCCUGCGAUUCUCCACCCU
2286
4197
CCCUGCGAUUCUCCACCCU
2286
4215
AGGGUGGAGAAUCGCAGGG
2581

4215
UUUCCCCUUCUUUCGUCCU
2287
4215
UUUCCCCUUCUUUCGUCCU
2287
4233
AGGACGAAAGAAGGGGAAA
2582

4233
UCACCAUCUCUGCAGACCC
2288
4233
UCACCAUCUCUGCAGACCC
2288
4251
GGGUCUGCAGAGAUGGUGA
2583

4251
CCUCUCCUCCUCCUUCCUC
2289
4251
CCUCUCCUCCUCCUUCCUC
2289
4269
GAGGAAGGAGGAGGAGAGG
2584

4269
CUUGGUCUCAGCACUGAUG
2290
4269
CUUGGUCUCAGCACUGAUG
2290
4287
CAUCAGUGCUGAGACCAAG
2585

4287
GGGAGGCUGGUGCCCAAGC
2291
4287
GGGAGGCUGGUGCCCAAGC
2291
4305
GCUUGGGCACCAGCCUCCC
2586

4305
CUGUGGCCUGCAGUCUGUG
2292
4305
CUGUGGCCUGCAGUCUGUG
2292
4323
CACAGACUGCAGGCCACAG
2587

4323
GAGGAGGGCUGUCUUGCCU
2293
4323
GAGGAGGGCUGUCUUGCCU
2293
4341
AGGCAAGACAGCCCUCCUC
2588

4341
UCACACUCCUCACAGCCUA
2294
4341
UCACACUCCUCACAGCCUA
2294
4359
UAGGCUGUGAGGAGUGUGA
2589

4359
ACUUCCCCUUCCCCGGGGC
2295
4359
ACUUCCCCUUCCCCGGGGC
2295
4377
GCCCCGGGGAAGGGGAAGU
2590

4377
CUGAGAGGGUGAAAGUGUG
2296
4377
CUGAGAGGGUGAAAGUGUG
2296
4395
CACACUUUCACCCUCUCAG
2591

4395
GUGGGGAAGGAGAGGACUG
2297
4395
GUGGGGAAGGAGAGGACUG
2297
4413
CAGUCCUCUCCUUCCCCAC
2592

4413
GGUUUCCUGGGUUCUCAGG
2298
4413
GGUUUCCUGGGUUCUCAGG
2298
4431
CCUGAGAACCCAGGAAACC
2593

4431
GGGCCAGGAGGAGUAACAG
2299
4431
GGGCCAGGAGGAGUAACAG
2299
4449
CUGUUACUCCUCCUGGCCC
2594

4449
GAACCAGGUCUGCUCCCCA
2300
4449
GAACCAGGUCUGCUCCCCA
2300
4467
UGGGGAGCAGACCUGGUUC
2595

4467
ACCUUACUCGGAUGGCCUC
2301
4467
ACCUUACUCGGAUGGCCUC
2301
4485
GAGGCCAUCCGAGUAAGGU
2596

4485
CCCUGCCCCUCUGCUGGCA
2302
4485
CCCUGCCCCUCUGCUGGCA
2302
4503
UGCCAGCAGAGGGGCAGGG
2597

4503
ACAGCCUGGGCAAGGGGAG
2303
4503
ACAGCCUGGGCAAGGGGAG
2303
4521
CUCCCCUUGCCCAGGCUGU
2598

4521
GAAGGUGGUCCCUGCAGAG
2304
4521
GAAGGUGGUCCCUGCAGAG
2304
4539
CUCUGCAGGGACCACCUUC
2599

4539
GGGGCUCCAGGCUGGUGAG
2305
4539
GGGGCUCCAGGCUGGUGAG
2305
4557
CUCACCAGCCUGGAGCCCC
2600

4557
GAGCCCCCCUGCUGUCAGG
2306
4557
GAGCCCCCCUGCUGUCAGG
2306
4575
CCUGACAGCAGGGGGGCUC
2601

4575
GACCAGAUUUUCCCAGCCA
2307
4575
GACCAGAUUUUCCCAGCCA
2307
4593
UGGCUGGGAAAAUCUGGUC
2602

4593
AUCCAGCAUGCUGCGGGGA
2308
4593
AUCCAGCAUGCUGCGGGGA
2308
4611
UCCCCGCAGCAUGCUGGAU
2603

4611
AGAAGGGGCAGAGGCUCAC
2309
4611
AGAAGGGGCAGAGGCUCAC
2309
4629
GUGAGCCUCUGCCCCUUCU
2604

4629
CCUCCCUCCUGGGGCCUUU
2310
4629
CCUCCCUCCUGGGGCCUUU
2310
4647
AAAGGCCCCAGGAGGGAGG
2605

4647
UUGUUUUGGAUCCUGGGGA
2311
4647
UUGUUUUGGAUCCUGGGGA
2311
4665
UCCCCAGGAUCCAAAACAA
2606

4665
AUGGUGAGAAUGGAGGUUC
2312
4665
AUGGUGAGAAUGGAGGUUC
2312
4683
GAACCUCCAUUCUCACCAU
2607

4683
CUAGAAGGGGUAAGGCCAG
2313
4683
CUAGAAGGGGUAAGGCCAG
2313
4701
CUGGCCUUACCCCUUCUAG
2608

4701
GAACCCAGGGAUCCAGGAG
2314
4701
GAACCCAGGGAUCCAGGAG
2314
4719
CUCCUGGAUCCCUGGGUUC
2609

4719
GUCGGCUCUCAGCUGGAGC
2315
4719
GUCGGCUCUCAGCUGGAGC
2315
4737
GCUCCAGCUGAGAGCCGAC
2610

4737
CUUCCAUACCUUCUGGGCU
2316
4737
CUUCCAUACCUUCUGGGCU
2316
4755
AGCCCAGAAGGUAUGGAAG
2611

4755
UCCCUUUGCUGACCACCAG
2317
4755
UCCCUUUGCUGACCACCAG
2317
4773
CUGGUGGUCAGCAAAGGGA
2612

4773
GCCCAAGGGAGCUAAGACC
2318
4773
GCCCAAGGGAGCUAAGACC
2318
4791
GGUCUUAGCUCCCUUGGGC
2613

4791
CAGGAGGGGGCUGGGCGCU
2319
4791
CAGGAGGGGGCUGGGCGCU
2319
4809
AGCGCCCAGCCCCCUCCUG
2614

4809
UGUCCCUUCUCUUUCCCAG
2320
4809
UGUCCCUUCUCUUUCCCAG
2320
4827
CUGGGAAAGAGAAGGGACA
2615

4827
GGAGCCCUGCCAGGGGCUG
2321
4827
GGAGCCCUGCCAGGGGCUG
2321
4845
CAGCCCCUGGCAGGGCUCC
2616

4845
GUGGGCCUACAAGGCUUCC
2322
4845
GUGGGCCUACAAGGCUUCC
2322
4863
GGAAGCCUUGUAGGCCCAC
2617

4863
CAGGGGAUGCCAUCCAGCC
2323
4863
CAGGGGAUGCCAUCCAGCC
2323
4881
GGCUGGAUGGCAUCCCCUG
2618

4881
CUGUAGGAAACCAAAGAUG
2324
4881
CUGUAGGAAACCAAAGAUG
2324
4899
CAUCUUUGGUUUCCUACAG
2619

4899
GGGAAGUGGCUCCUAGGGG
2325
4899
GGGAAGUGGCUCCUAGGGG
2325
4917
CCCCUAGGAGCCACUUCCG
2620

4917
GGCUGACUCUUCCUUCCUC
2326
4917
GGCUGAGUCUUCCUUCCUC
2326
4935
GAGGAAGGAAGAGUCAGCC
2621

4935
CCUCCUCCCCAGUACCACA
2327
4935
CCUCCUCCCCAGUACCACA
2327
4953
UGUGGUACUGGGGAGGAGG
2622

4953
AUAUACUUUCUCUCCUUCU
2328
4953
AUAUACUUUCUCUCCUUCU
2328
4971
AGAAGGAGAGAAAGUAUAU
2623

4971
UAUCUCCAGGGCCCCACCA
2329
4971
UAUCUCCAGGGCCCCACCA
2329
4989
UGGUGGGGCCCUGGAGAUA
2624

4989
AAUCUGUUUACAUAUUUAU
2330
4989
AAUCUGUUUACAUAUUUAU
2330
5007
AUAAAUAUGUAAACAGAUU
2625

5007
UUAUCCUAUGGGGGCCUGA
2331
5007
UUAUCCUAUGGGGGCCUGA
2331
5025
UCAGGCCCCCAUAGGAUAA
2626

5025
AGCAGGAUUGAGGGAGCCA
2332
5025
AGCAGGAUUGAGGGAGCCA
2332
5043
UGGCUCCCUCAAUCCUGCU
2627

5043
AGGGGAGGGGCAGGAGUCC
2333
5043
AGGGGAGGGGCAGGAGUCC
2333
5061
GGACUCCUGCCCCUCCCCU
2628

5061
CCAGCACCAUCGGUUCAUA
2334
5061
CCAGCACCAUCGGUUCAUA
2334
5079
UAUGAACCGAUGGUGCUGG
2629

5079
AGUGUGCUUGUGUGUUUGU
2335
5079
AGUGUGCUUGUGUGUUUGU
2335
5097
ACAAACACACAAGCACACU
2630

5097
UUUUAGAUCCUCCUGGGGG
2336
5097
UUUUAGAUCCUCCUGGGGG
2336
5115
CCCCCAGGAGGAUCUAAAA
2631

5115
GAUGGGGAUGGGGCCAGGC
2337
5115
GAUGGGGAUGGGGCCAGGC
2337
5133
GCCUGGCCCCAUCCCCAUC
2632

5133
CUCAGUGUACUAGGCCUCU
2338
5133
CUCAGUGUACUAGGCCUCU
2338
5151
AGAGGCCUAGUACACUGAG
2633

5151
UCUGUGCUGAGCCCCAGGC
2339
5151
UCUGUGCUGAGCCCCAGGC
2339
5169
GCCUGGGGCUCAGCACAGA
2634

5169
CUCCCGGCCCCUUACCCAC
2340
5169
CUCCCGGCCCCUUACCCAC
2340
5187
GUGGGUAAGGGGCCGGGAG
2635

5187
CUCUCUCCCUGUGGCUGGU
2341
5187
CUCUCUCCCUGUGGCUGGU
2341
5205
ACCAGCCACAGGGAGAGAG
2636

5205
UCUGGUUCUCAUGUAAACC
2342
5205
UCUGGUUCUCAUGUAAACC
2342
5223
GGUUUACAUGAGAACCAGA
2637

5223
CCACUCCUUGCUUUGUCUC
2343
5223
CCACUCCUUGCUUUGUCUC
2343
5241
GAGACAAAGCAAGGAGUGG
2638

5241
CCCUGGAUAUGGAUUUCAG
2344
5241
CCCUGGAUAUGGAUUUCAG
2344
5259
CUGAAAUCCAUAUCCAGGG
2639

5259
GUUAAGUAUUUUGUAACCC
2345
5259
GUUAAGUAUUUUGUAACCC
2345
5277
GGGUUACAAAAUACUUAAC
2640

5277
CGUUACACUGUGUGUCCUU
2346
5277
CGUUACACUGUGUGUCCUU
2346
5295
AAGGACACACAGUGUAACG
2641

5295
UGUGUAAAUAAACUUGUUU
2347
5295
UGUGUAAAUAAACUUGUUU
2347
5313
AAACAAGUUUAUUUACACA
2642

HDAC6: NM_006044.2

3
GCAGUCCCCUGAGGAGCGG
2755
3
GGAGUCCCCUGAGGAGCGG
2755
21
CCGCUCCUCAGGGGACUGC
2982

21
GGGCUGGUUGAAACGCUAG
2756
21
GGGCUGGUUGAAACGCUAG
2756
39
CUAGCGUUUCAACCAGCCC
2983

39
GGGGCGGGAUCUGGCGGAG
2757
39
GGGGCGGGAUCUGGCGGAG
2757
57
CUCCGCCAGAUCCCGCCCC
2984

57
GUGGAAGAACCGCGGCAGG
2758
57
GUGGAAGAACGGCGGCAGG
2758
75
CCUGCCGCGGUUGUUCCAC
2985

75
GGGCCAAGCCUCCUCAACU
2759
75
GGGCCAAGCCUCCUCAACU
2759
93
AGUUGAGGAGGCUUGGCCC
2986

93
UAUGACCUCAACCGGCCAG
2760
93
UAUGACCUCAACCGGCCAG
2760
111
CUGGCCGGUUGAGGUCAUA
2987

111
GGAUUCCACCACAACCAGG
2761
111
GGAUUCCACCACAACCAGG
2761
129
CCUGGUUGUGGUGGAAUCC
2988

129
GCAGCGAAGAAGUAGGCAG
2762
129
GCAGCGAAGAAGUAGGCAG
2762
147
CUGCCUACUUCUUCGCUGC
2989

147
GAACCCCCAGUCGCCCCCU
2763
147
GAACCCCCAGUCGCCCCCU
2763
165
AGGGGGCGACUGGGGGUUC
2990

165
UCAGGACUCCAGUGUCACU
2764
165
UCAGGACUCCAGUGUCACU
2764
183
AGUGACACUGGAGUCCUGA
2991

183
UUCGAAGCGAAAUAUUAAA
2765
183
UUCGAAGCGAAAUAUUAAA
2765
201
UUUAAUAUUUCGCUUCGAA
2992

201
AAAGGGAGCCGUUCCCCGC
2766
201
AAAGGGAGCCGUUCCCCGC
2766
219
GCGGGGAACGGCUCCCUUU
2993

219
CUCUAUCCCCAAUCUAGCG
2767
219
CUCUAUCCCCAAUCUAGCG
2767
237
CGCUAGAUUGGGGAUAGAG
2994

237
GGAGGUAAAGAAGAAAGGC
2768
237
GGAGGUAAAGAAGAAAGGC
2768
255
GCCUUUCUUCUUUACCUCC
2995

255
CAAAAUGAAGAAGCUCGGC
2769
255
CAAAAUGAAGAAGCUCGGC
2769
273
GCCGAGCUUCUUCAUUUUG
2996

273
CCAAGCAAUGGAAGAAGAC
2770
273
CCAAGCAAUGGAAGAAGAC
2770
291
GUCUUCUUCCAUUGCUUGG
2997

291
CCUAAUCGUGGGACUGCAA
2771
291
CCUAAUCGUGGGACUGCAA
2771
309
UUGCAGUCCCACGAUUAGG
2998

309
AGGGAUGGAUCUGAACCUU
2772
309
AGGGAUGGAUCUGAACCUU
2772
327
AAGGUUCAGAUCCAUCCCU
2999

327
UGAGGCUGAAGCACUGGCU
2773
327
UGAGGCUGAAGCACUGGCU
2773
345
AGCCAGUGCUUCAGCCUCA
3000

345
UGGCACUGGCUUGGUGUUG
2774
345
UGGCACUGGCUUGGUGUUG
2774
363
CAACACCAAGCCAGUGCCA
3001

363
GGAUGAGCAGUUAAAUGAA
2775
363
GGAUGAGCAGUUAAAUGAA
2775
381
UUCAUUUAACUGCUCAUCC
3002

381
AUUCCAUUGCCUCUGGGAU
2776
381
AUUCCAUUGCCUCUGGGAU
2776
399
AUCCCAGAGGCAAUGGAAU
3003

399
UGACAGCUUCCCGGAAGGC
2777
399
UGACAGCUUCCCGGAAGGC
2777
417
GCCUUCCGGGAAGCUGUCA
3004

417
CCCUGAGCGGCUCCAUGCC
2778
417
CCCUGAGCGGCUCCAUGCC
2778
435
GGCAUGGAGCCGCUCAGGG
3005

435
CAUCAAGGAGCAACUGAUC
2779
435
CAUCAAGGAGCAACUGAUC
2779
453
GAUCAGUUGCUCCUUGAUG
3006

453
CCAGGAGGGCCUCCUAGAU
2780
453
CCAGGAGGGCCUCCUAGAU
2780
471
AUCUAGGAGGCCCUCCUGG
3007

471
UCGCUGCGUGUCCUUUCAG
2781
471
UCGCUGCGUGUCCUUUCAG
2781
489
CUGAAAGGACACGCAGCGA
3008

489
GGCCCGGUUUGCUGAAAAG
2782
489
GGCCCGGUUUGCUGAAAAG
2782
507
CUUUUCAGCAAACCGGGCC
3009

507
GGAAGAGCUGAUGUUGGUU
2783
507
GGAAGAGCUGAUGUUGGUU
2783
525
AACCAACAUCAGCUCUUCC
3010

525
UCACAGCCUAGAAUAUAUU
2784
525
UCACAGCCUAGAAUAUAUU
2784
543
AAUAUAUUCUAGGCUGUGA
3011

543
UGAUCUGAUGGAAACAACC
2785
543
UGAUCUGAUGGAAACAACC
2785
561
GGUUGUUUCCAUCAGAUCA
3012

561
CCAGUACAUGAAUGAGGGA
2786
561
CCAGUACAUGAAUGAGGGA
2786
579
UCCCUCAUUCAUGUACUGG
3013

579
AGAACUCCGUGUCCUAGCA
2787
579
AGAACUCCGUGUCCUAGCA
2787
597
UGCUAGGACACGGAGUUCU
3014

597
AGACACCUACGACUCAGUU
2788
597
AGACACCUACGACUCAGUU
2788
615
AACUGAGUCGUAGGUGUCU
3015

615
UUAUCUGCAUCCGAACUCA
2789
615
UUAUCUGCAUCCGAACUCA
2789
633
UGAGUUCGGAUGCAGAUAA
3016

633
AUACUCCUGUGGCUGCCUG
2790
633
AUACUCCUGUGCCUGCCUG
2790
651
CAGGCAGGCACAGGAGUAU
3017

651
GGCCUCAGGCUCUGUCGUC
2791
651
GGCCUCAGGCUCUGUCCUC
2791
669
GAGGACAGAGCCUGAGGCC
3018

669
CAGGCUGGUGGAUGCGGUC
2792
669
CAGGCUGGUGGAUGCGGUC
2792
687
GACCGCAUCCACCAGCCUG
3019

687
CCUGGGGGCUGAGAUCCGG
2793
687
CCUGGGGGCUGAGAUCCGG
2793
705
CCGGAUCUCAGCCCCCAGG
3020

705
GAAUGGCAUGGCCAUCAUU
2794
705
GAAUGGCAUGGCCAUCAUU
2794
723
AAUGAUGGCCAUGCGAUUC
3021

723
UAGGCCUCCUGGACAUCAC
2795
723
UAGGCCUCCUGGACAUCAC
2795
741
GUGAUGUCCAGGAGGCCUA
3022

741
CGCCCAGCACAGUCUUAUG
2796
741
CGCCCAGCACAGUCUUAUG
2796
759
CAUAAGACUGUGCUGGGCG
3023

759
GGAUGGCUAUUGCAUGUUC
2797
759
GGAUGGCUAUUGCAUGUUC
2797
777
GAACAUGCAAUAGCCAUCC
3024

777
CAACCACGUGGCUGUGGCA
2798
777
CAACCACGUGGCUGUGGCA
2798
795
UGCCACAGCCACGUGGUUG
3025

795
AGCCCGCUAUGCUCAACAG
2799
795
AGCCCGCUAUGCUCAACAG
2799
813
CUGUUGAGCAUAGCGGGCU
3026

813
GAAACACCGCAUCCGGAGG
2800
813
GAAACACCGCAUCCGGAGG
2800
831
CCUCCGGAUGCGGUGUUUC
3027

831
GGUCCUUAUCGUAGAUUGG
2801
831
GGUCCUUAUCGUAGAUUGG
2801
849
CCAAUCUACGAUAAGGACC
3028

849
GGAUGUGCACCACGGUCAA
2802
849
GGAUGUGCACCACGGUCAA
2802
867
UUGACCGUGGUGCACAUCC
3029

867
AGGAACACAGUUCACCUUC
2803
867
AGGAACACAGUUCACCUUC
2803
885
GAAGGUGAACUGUGUUCCU
3030

885
CGACCAGGACCCCAGUGUC
2804
885
CGACCAGGACCCCAGUGUC
2804
903
GACACUGGGGUCCUGGUCG
3031

903
CCUCUAUUUCUCCAUCCAC
2805
903
CCUCUAUUUCUCCAUCCAC
2805
921
GUGGAUGGAGAAAUAGAGG
3032

921
CCGCUACGAGCAGGGUAGG
2806
921
CCGCUACGAGCAGGGUAGG
2806
939
CCUACCCUGCUCGUAGCGG
3033

939
GUUCUGGGCCCACCUGAAG
2807
939
GUUCUGGCCCCACCUGAAG
2807
957
CUUCAGGUGGGGCCAGAAC
3034

957
GGCCUCUAACUGGUCCACC
2808
957
GGCCUCUAACUGGUCCACC
2808
975
GGUGGACCAGUUAGAGGCC
3035

975
CACAGGUUUCGGCCAAGGC
2809
975
CACAGGUUUCGGCCAAGGC
2809
993
GCCUUGGCCGAAACCUGUG
3036

993
CCAAGGAUAUACCAUCAAU
2810
993
CCAAGGAUAUACCAUCAAU
2810
1011
AUUGAUGGUAUAUCCUUGG
3037

1011
UGUGCCUUGGAACCAGGUG
2811
1011
UGUGCCUUGGAACCAGGUG
2811
1029
CACCUGGUUCCAAGGCACA
3038

1029
GGGGAUGCGGGAUGCUGAC
2812
1029
GGGGAUGCGGGAUGCUGAC
2812
1047
GUCAGCAUCCCGCAUCCCC
3039

1047
CUACAUUGCUGCUUUCCUG
2813
1047
CUACAUUGCUGCUUUCCUG
2813
1065
CAGGAAAGCAGCAAUGUAG
3040

1065
GCACGUCCUGCUGCCAGUC
2814
1065
GCACGUCCUGCUGCCAGUC
2814
1083
GACUGGCAGCAGGACGUGC
3041

1083
CGCCCUCGAGUUCCAGCCU
2815
1083
CGCCCUCGAGUUCCAGCCU
2815
1101
AGGCUGGAACUCGAGGGCG
3042

1101
UCAGCUGGUCCUGGUGGCU
2816
1101
UCAGCUGGUCCUGGUGGCU
2816
1119
AGCCACCAGGACCAGCUGA
3043

1119
UGCUGGAUUUGAUGCCCUG
2817
1119
UGCUGGAUUUGAUGCCCUG
2817
1137
CAGGGCAUCAAAUCCAGCA
3044

1137
GCAAGGGGACCCCAAGGGU
2818
1137
GCAAGGGGACCCCAAGGGU
2818
1155
ACCCUUGGGGUCCCCUUGC
3045

1155
UGAGAUGGCCGCCACUCCG
2819
1155
UGAGAUGGCCGCCACUCCG
2819
1173
CGGAGUGGCGGCCAUCUCA
3046

1173
GGCAGGGUUCGCCCAGCUA
2820
1173
GGCAGGGUUCGCCCAGCUA
2820
1191
UAGCUGGGCGAACCCUGCC
3047

1191
AACCCACCUGCUCAUGGGU
2821
1191
AACCCACCUGCUCAUGGGU
2821
1209
ACCCAUGAGCAGGUGGGUU
3048

1209
UCUGGCAGGAGGCAAGCUG
2822
1209
UCUGGCAGGAGGCAAGCUG
2822
1227
CAGCUUGCCUCCUGCCAGA
3049

1227
GAUCCUGUCUCUGGAGGGU
2823
1227
GAUCCUGUCUCUGGAGGGU
2823
1245
ACCCUCCAGAGACAGGAUC
3050

1245
UGGCUACAACCUCCGCGCC
2824
1245
UGGCUACAACCUCCGCGCC
2824
1263
GGCGCGGAGGUUGUAGCCA
3051

1263
CCUGGCUGAAGGCGUCAGU
2825
1263
CCUGGCUGAAGGCGUCAGU
2825
1281
ACUGACGCCUUCAGCCAGG
3052

1281
UGCUUCGCUCCACACCCUU
2826
1281
UGCUUCGCUCCACACCCUU
2826
1299
AAGGGUGUGGAGCGAAGCA
3053

1299
UCUGGGAGACCCUUGCCCC
2827
1299
UCUGGGAGACCCUUGCCCC
2827
1317
GGGGCAAGGGUCUCCCAGA
3054

1317
CAUGCUGGAGUCACCUGGU
2828
1317
CAUGGUGGAGUCACCUGGU
2828
1335
ACCAGGUGACUCCAGGAUG
3055

1335
UGCCCCCUGCCGGAGUGCC
2829
1335
UGCCCCCUGCCGGAGUGCC
2829
1353
GGCACUCCGGCAGGGGGCA
3056

1353
CCAGGCUUCAGUUUCCUGU
2830
1353
CCAGGCUUCAGUUUCCUGU
2830
1371
ACAGGAAACUGAAGCCUGG
3057

1371
UGCUCUGGAAGCCCUUGAG
2831
1371
UGCUCUGGAAGCCCUUGAG
2831
1389
CUCAAGGGCUUCCAGAGCA
3058

1389
GCCCUUCUGGGAGGUUCUU
2832
1389
GCCCUUCUGGGAGGUUCUU
2832
1407
AAGAACCUCCCAGAAGGGC
3059

1407
UGUGAGAUCAACUGAGACC
2833
1407
UGUGAGAUCAACUGAGACC
2833
1425
GGUCUCAGUUGAUCUCACA
3060

1425
CGUGGAGAGGGACAACAUG
2834
1425
CGUGGAGAGGGACAACAUG
2834
1443
CAUGUUGUCCCUCUCCACG
3061

1443
GGAGGAGGACAAUGUAGAG
2835
1443
GGAGGAGGACAAUGUAGAG
2835
1461
CUCUACAUUGUCCUCCUCC
3062

1461
GGAGAGCGAGGAGGAAGGA
2836
1461
GGAGAGCGAGGAGGAAGGA
2836
1479
UCCUUCCUCCUCGCUCUCC
3063

1479
ACCCUGGGAGCCCCCUGUG
2837
1479
ACCCUGGGAGCCCCCUGUG
2837
1497
CACAGGGGGCUCCCAGGGU
3064

1497
GCUCCCAAUCCUGACAUGG
2838
1497
GCUCCCAAUCCUGACAUGG
2838
1515
CCAUGUCAGGAUUGGGAGC
3065

1515
GCCAGUGCUACAGUCUCGC
2839
1515
GCCAGUGCUACAGUCUCGC
2839
1533
GCGAGACUGUAGCACUGGC
3066

1533
CACAGGGCUGGUCUAUGAC
2840
1533
CACAGGGCUGGUCUAUGAC
2840
1551
GUCAUAGACCAGCCCUGUG
3067

1551
CCAAAAUAUGAUGAAUCAC
2841
1551
CCAAAAUAUGAUGAAUCAC
2841
1569
GUGAUUCAUCAUAUUUUGG
3068

1569
CUGCAACUUGUGGGACAGC
2842
1569
CUGCAACUUGUGGGACAGC
2842
1587
GCUGUCCCACAAGUUGCAG
3069

1587
CCACCACCCUGAGGUACCC
2843
1587
CCACCACCCUGAGGUACCC
2843
1605
GGGUACCUCAGGGUGGUGG
3070

1605
CCAGCGCAUCUUGCGGAUC
2844
1605
CCAGCGCAUCUUGCGGAUC
2844
1623
GAUCCGCAAGAUGCGCUGG
3071

1623
CAUGUGCCGUCUGGAGGAG
2845
1623
CAUGUGCCGUCUGGAGGAG
2845
1641
CUCCUCCAGACGGCACAUG
3072

1641
GCUGGGCCUUGCCGGGCGC
2846
1641
GCUGGGCCUUGCCGGGCGC
2846
1659
GCGCCCGGCAAGGCCCAGC
3073

1659
CUGCCUCACCCUGACACCG
2847
1659
CUGCCUCACCCUGACACCG
2847
1677
CGGUGUCAGGGUGAGGCAG
3074

1677
GCGCCCUGCCACAGAGGCU
2848
1677
GCGCCCUGCCACAGAGGCU
2848
1695
AGCCUCUGUGGCAGGGCGC
3075

1695
UGAGCUGCUCACCUGUCAC
2849
1695
UGAGCUGCUCACCUGUCAC
2849
1713
GUGACAGGUGAGCAGCUCA
3076

1713
CAGUGCUGAGUACGUGGGU
2850
1713
CAGUGCUGAGUACGUGGGU
2850
1731
ACCCACGUACUCAGCACUG
3077

1731
UCAUCUCCGGGCCACAGAG
2851
1731
UCAUCUCCGGGCCACAGAG
2851
1749
CUCUGUGGCCCGGAGAUGA
3078

1749
GAAAAUGAAAACCCGGGAG
2852
1749
GAAAAUGAAAACCCGGGAG
2852
1767
CUCCCGGGUUUUCAUUUUC
3079

1767
GCUGCACCGUGAGAGUUCC
2853
1767
GCUGCACCGUGAGAGUUCC
2853
1785
GGAACUCUCACGGUGCAGC
3080

1785
CAACUUUGACUCCAUCUAU
2854
1785
CAACUUUGACUCCAUCUAU
2854
1803
AUAGAUGGAGUCAAAGUUG
3081

1803
UAUCUGCCCCAGUACCUUC
2855
1803
UAUCUGCCCCAGUACCUUC
2855
1821
GAAGGUACUGGGGCAGAUA
3082

1821
CGCCUGUGCACAGCUUGCC
2856
1821
CGCCUGUGCACAGCUUGCC
2856
1839
GGCAAGCUGUGCACAGGCG
3083

1839
CACUGGCGCUGCCUGCCGC
2857
1839
CACUGGCGCUGCCUGCCGC
2857
1857
GCGGCAGGCAGCGCCAGUG
3084

1857
CCUGGUGGAGGCUGUGCUC
2858
1857
CCUGGUGGAGGCUGUGCUC
2858
1875
GAGCACAGCCUCCACCAGG
3085

1875
CUCAGGAGAGGUUCUGAAU
2859
1875
CUCAGGAGAGGUUCUGAAU
2859
1893
AUUCAGAACCUCUCCUGAG
3086

1893
UGGUGCUGCUGUGGUGCGU
2860
1893
UGGUGCUGCUGUGGUGCGU
2860
1911
ACGCACCACAGCAGCACCA
3087

1911
UCCCCCAGGACACCACGCA
2861
1911
UCCCCCAGGACACCACGCA
2861
1929
UGCGUGGUGUCCUGGGGGA
3088

1929
AGAGCAGGAUGCAGCUUGC
2862
1929
AGAGCAGGAUGCAGCUUGC
2862
1947
GCAAGCUGCAUCCUGCUCU
3089

1947
CGGUUUUUGCUUUUUCAAC
2863
1947
CGGUUUUUGCUUUUUCAAC
2863
1965
GUUGAAAAAGCAAAAACCG
3090

1965
CUCUGUGGCUGUGGCUGCU
2864
1965
CUCUGUGGCUGUGGCUGCU
2864
1983
AGCAGCCACAGCCACAGAG
3091

1983
UCGCCAUGCCCAGACUAUC
2865
1983
UCGCCAUGCCCAGACUAUC
2865
2001
GAUAGUCUGGGCAUGGCGA
3092

2001
CAGUGGGCAUGCCCUACGG
2866
2001
CAGUGGGCAUGCCCUACGG
2866
2019
CCGUAGGGCAUGCCCACUG
3093

2019
GAUCCUGAUUGUGGAUUGG
2867
2019
GAUCCUGAUUGUGGAUUGG
2867
2037
CCAAUCCACAAUCAGGAUC
3094

2037
GGAUGUCCACCACGGUAAU
2868
2037
GGAUGUCCACCACGGUAAU
2868
2055
AUUACCGUGGUGGACAUCC
3095

2055
UGGAACUCAGCACAUGUUU
2869
2055
UGGAACUCAGCACAUGUUU
2869
2073
AAACAUGUGCUGAGUUCCA
3096

2073
UGAGGAUGACCCCAGUGUG
2870
2073
UGAGGAUGACCCCAGUGUG
2870
2091
CACACUGGGGUCAUCCUCA
3097

2091
GCUAUAUGUGUCCCUGCAC
2871
2091
GCUAUAUGUGUCCCUGCAC
2871
2109
GUGCAGGGACACAUAUAGC
3098

2109
CCGCUAUGAUCAUGGCACC
2872
2109
CCGCUAUGAUCAUGGCACC
2872
2127
GGUGCCAUGAUCAUAGCGG
3099

2127
CUUCUUCCCCAUGGGGGAU
2873
2127
CUUCUUCCCCAUGGGGGAU
2873
2145
AUCCCCCAUGGGGAAGAAG
3100

2145
UGAGGGUGCCAGCAGCCAG
2874
2145
UGAGGGUGCCAGCAGCCAG
2874
2163
CUGGCUGCUGGCACCCUCA
3101

2163
GAUCGGCCGGGCUGCGGGC
2875
2163
GAUCGGCCGGGCUGCGGGC
2875
2181
GCCCGCAGCCCGGCCGAUC
3102

2181
CACAGGCUUCACCGUCAAC
2876
2181
CACAGGCUUCACCGUCAAC
2876
2199
GUUGACGGUGAAGCCUGUG
3103

2199
CGUGGCAUGGAACGGGCCC
2877
2199
CGUGGCAUGGAACGGGCCC
2877
2217
GGGCCCGUUCCAUGCCACG
3104

2217
CCGCAUGGGUGAUGCUGAC
2878
2217
CCGCAUGGGUGAUGCUGAC
2878
2235
GUCAGCAUCACCCAUGCGG
3105

2235
CUACCUAGCUGCCUGGCAU
2879
2235
CUACCUAGCUGCCUGGCAU
2879
2253
AUGCCAGGCAGCUAGGUAG
3106

2253
UCGCCUGGUGCUUCCCAUU
2880
2253
UCGCCUGGUGCUUCCCAUU
2880
2271
AAUGGGAAGCACCAGGCGA
3107

2271
UGCCUACGAGUUUAACCCA
2881
2271
UGCCUACGAGUUUAACCCA
2881
2289
UGGGUUAAACUCGUAGGCA
3108

2289
AGAACUGGUGCUGGUCUCA
2882
2289
AGAACUGGUGCUGGUCUCA
2882
2307
UGAGACCAGCACCAGUUCU
3109

2307
AGCUGGCUUUGAUGCUGCA
2883
2307
AGCUGGCUUUGAUGCUGCA
2883
2325
UGCAGCAUCAAAGCCAGCU
3110

2325
ACGGGGGGAUCCGCUGGGG
2884
2325
ACGGGGGGAUCCGCUGGGG
2884
2343
CCCCAGCGGAUCCCCCCGU
3111

2343
GGGCUGCCAGGUGUCACCU
2885
2343
GGGCUGCCAGGUGUCACCU
2885
2361
AGGUGACACCUGGCAGCCC
3112

2361
UGAGGGUUAUGCCCACCUC
2886
2361
UGAGGGUUAUGCCCACCUC
2886
2379
GAGGUGGGCAUAACCCUCA
3113

2379
CACCCACCUGCUGAUGGGC
2887
2379
CACCCACCUGCUGAUGGGC
2887
2397
GCCCAUCAGCAGGUGGGUG
3114

2397
CCUUGCCAGUGGCCGCAUU
2888
2397
CCUUGCCAGUGGCCGCAUU
2888
2415
AAUGCGGCCACUGGCAAGG
3115

2415
UAUCCUUAUCCUAGAGGGU
2889
2415
UAUCCUUAUCCUAGAGGGU
2889
2433
ACCCUCUAGGAUAAGGAUA
3116

2433
UGGCUAUAACCUGACAUCC
2890
2433
UGGCUAUAACCUGACAUCC
2890
2451
GGAUGUCAGGUUAUAGCCA
3117

2451
CAUCUCAGAGUCCAUGGCU
2891
2451
CAUCUCAGAGUCCAUGGCU
2891
2469
AGCCAUGGACUCUGAGAUG
3118

2469
UGCCUGCACUCGCUCCCUC
2892
2469
UGCCUGCACUCGCUCCCUC
2892
2487
GAGGGAGCGAGUGCAGGCA
3119

2487
CCUUGGAGACCGACCACCC
2893
2487
CCUUGGAGACCCACCACCC
2893
2505
GGGUGGUGGGUCUCCAAGG
3120

2505
CCUGCUGACCCUGCCACGG
2894
2505
CCUGCUGACCCUGCCACGG
2894
2523
CCGUGGCAGGGUCAGCAGG
3121

2523
GCCCCCACUAUCAGGGGCC
2895
2523
GCCCCCACUAUCAGGGGCC
2895
2541
GGCCCCUGAUAGUGGGGGC
3122

2541
CCUGGCCUCAAUCACUGAG
2896
2541
CCUGGCCUCAAUCACUGAG
2896
2559
CUCAGUGAUUGAGGCCAGG
3123

2559
GACCAUCCAAGUCCAUCGC
2897
2559
GACCAUCCAAGUCCAUCGC
2897
2577
GCGAUGGACUUGGAUGGUC
3124

2577
CAGAUACUGGCGCAGCUUA
2898
2577
CAGAUACUGGCGCAGCUUA
2898
2595
UAAGCUGCGCCAGUAUCUG
3125

2595
ACGGGUCAUGAAGGUAGAA
2899
2595
ACGGGUCAUGAAGGUAGAA
2899
2613
UUCUACCUUCAUGACCCGU
3126

2613
AGACAGAGAAGGACCCUCC
2900
2613
AGACAGAGAAGGACCCUCC
2900
2631
GGAGGGUCCUUCUCUGUCU
3127

2631
CAGUUCUAAGUUGGUCACC
2901
2631
CAGUUCUAAGUUGGUCACC
2901
2649
GGUGACCAACUUAGAACUG
3128

2649
CAAGAAGGCACCCCAACCA
2902
2649
CAAGAAGGCACCCCAACCA
2902
2667
UGGUUGGGGUGCCUUCUUG
3129

2667
AGCCAAACCUAGGUUAGCU
2903
2667
AGCCAAACCUAGGUUAGCU
2903
2685
AGCUAACCUAGGUUUGGCU
3130

2685
UGAGCGGAUGACCACACGA
2904
2685
UGAGCGGAUGACCACACGA
2904
2703
UCGUGUGGUCAUCCGCUCA
3131

2703
AGAAAAGAAGGUUCUGGAA
2905
2703
AGAAAAGAAGGUUCUGGAA
2905
2721
UUCCAGAACCUUCUUUUCU
3132

2721
AGCAGGCAUGGGGAAAGUC
2906
2721
AGCAGGCAUGGGGAAAGUC
2906
2739
GACUUUCCCCAUGCCUGCU
3133

2739
CACCUCGGCAUCAUUUGGG
2907
2739
CACCUCGGCAUCAUUUGGG
2907
2757
CCCAAAUGAUGCCGAGGUG
3134

2757
GGAAGAGUCCACUCCAGGC
2908
2757
GGAAGAGUCCACUCCAGGC
2908
2775
GCCUGGAGUGGACUCUUCC
3135

2775
CCAGACUAACUGAGAGACA
2909
2775
CCAGACUAACUCAGAGACA
2909
2793
UGUCUCUGAGUUAGUCUGG
3136

2793
AGCUGUGGUGGCCCUCACU
2910
2793
AGCUGUGGUGGCCCUCACU
2910
2811
AGUGAGGGCCACCACAGCU
3137

2811
UCAGGACCAGCCCUCAGAG
2911
2811
UCAGGACCAGCCCUCAGAG
2911
2829
CUCUGAGGGCUGGUCCUGA
3138

2829
GGCAGCCACAGGGGGAGCC
2912
2829
GGCAGCCACAGGGGGAGCC
2912
2847
GGCUCCCCCUGUGGCUGCC
3139

2847
CACUCUGGCCCAGACCAUU
2913
2847
CACUCUGGCCCAGACCAUU
2913
2865
AAUGGUCUGGGGCAGAGUG
3140

2865
UUCUGAGGCAGCCAUUGGG
2914
2865
UUCUGAGGCAGCCAUUGGG
2914
2883
CCCAAUGGCUGCCUCAGAA
3141

2883
GGGAGCCAUGCUGGGCCAG
2915
2883
GGGAGCCAUGGUGGGCCAG
2915
2901
CUGGCCCAGCAUGGCUCCC
3142

2901
GACCACCUCAGAGGAGGCU
2916
2901
GACCACCUCAGAGGAGGCU
2916
2919
AGCCUCCUCUGAGGUGGUC
3143

2919
UGUCGGGGGAGCCACUCCG
2917
2919
UGUCGGGGGAGCCACUCCG
2917
2937
CGGAGUGGCUCCCCCGACA
3144

2937
GGACCAGACCACCUCAGAG
2918
2937
GGACCAGACCACCUCAGAG
2918
2955
CUCUGAGGUGGUCUGGUCC
3145

2955
GGAGACUGUGGGAGGAGCC
2919
2955
GGAGACUGUGGGAGGAGCC
2919
2973
GGCUCCUCCCACAGUCUCC
3146

2973
CAUUCUGGACCAGACCACC
2920
2973
CAUUCUGGACCAGACCACC
2920
2991
GGUGGUCUGGUCCAGAAUG
3147

2991
CUCAGAGGAUGCUGUUGGG
2921
2991
CUCAGAGGAUGCUGUUGGG
2921
3009
CCCAACAGCAUCCUCUGAG
3148

3009
GGGAGCCACGCUGGGCCAG
2922
3009
GGGAGCCACGCUGGGCCAG
2922
3027
CUGGCCCAGCGUGGCUCCC
3149

3027
GACUACCUCAGAGGAGGCU
2923
3027
GACUACCUCAGAGGAGGCU
2923
3045
AGCCUCCUCUGAGGUAGUC
3150

3045
UGUAGGAGGAGCUACACUG
2924
3045
UGUAGGAGGAGGUACACUG
2924
3063
CAGUGUAGCUCCUCCUACA
3151

3063
GGCCCAGACCACCUCGGAG
2925
3063
GGCCCAGACCACCUCGGAG
2925
3081
CUCCGAGGUGGUCUGGGCC
3152

3081
GGCAGCCAUGGAGGGAGCC
2926
3081
GGCAGCCAUGGAGGGAGCC
2926
3099
GGCUCCCUCCAUGGCUGCC
3153

3099
CACACUGGACCAGACUACG
2927
3099
CACACUGGACCAGACUACG
2927
3117
CGUAGUCUGGUCCAGUGUG
3154

3117
GUCAGAGGAGGCUCCAGGG
2928
3117
GUCAGAGGAGGCUCCAGGG
2928
3135
CCCUGGAGCCUCCUCUGAC
3155

3135
GGGCACCGAGCUGAUCCAA
2929
3135
GGGCACCGAGCUGAUCCAA
2929
3153
UUGGAUCAGCUCGGUGCCC
3156

3153
AACUCCUCUAGCCUCGAGC
2930
3153
AACUCCUCUAGCCUCGAGC
2930
3171
GCUCGAGGCUAGAGGAGUU
3157

3171
CACAGACCACCAGACCCCC
2931
3171
CACAGACCACCAGACCCCC
2931
3189
GGGGGUCUGGUGGUCUGUG
3158

3189
CCCAACCUCACCUGUGCAG
2932
3189
CCCAACCUCACCUGUGCAG
2932
3207
CUGCACAGGUGAGGUUGGG
3159

3207
GGGAACUACACCCCAGAUA
2933
3207
GGGAACUACACCCCAGAUA
2933
3225
UAUCUGGGGUGUAGUUCCC
3160

3225
AUCUCCCAGUACACUGAUU
2934
3225
AUCUCCCAGUACACUGAUU
2934
3243
AAUCAGUGUACUGGGAGAU
3161

3243
UGGGAGUCUCAGGACCUUG
2935
3243
UGGGAGUCUCAGGACCUUG
2935
3261
CAAGGUCCUGAGACUCCCA
3162

3261
GGAGCUAGGCAGCGAAUCU
2936
3261
GGAGCUAGGCAGCGAAUCU
2936
3279
AGAUUCGCUGCCUAGCUCC
3163

3279
UCAGGGGGCCUCAGAAUCU
2937
3279
UCAGGGGGCCUCAGAAUCU
2937
3297
AGAUUCUGAGGCCCCCUGA
3164

3297
UCAGGCCCCAGGAGAGGAG
2938
3297
UCAGGCCCCAGGAGAGGAG
2938
3315
CUCCUCUCCUGGGGCCUGA
3165

3315
GAACCUACUAGGAGAGGCA
2939
3315
GAACCUACUAGGAGAGGCA
2939
3333
UGCCUCUCCUAGUAGGUUC
3166

3333
AGGUGGAGGUCAGGACAUG
2940
3333
AGCUGGAGGUCAGGACAUG
2940
3351
CAUGUCCUGACCUCCAGCU
3167

3351
GGCUGAUUCGAUGCUGAUG
2941
3351
GGCUGAUUCGAUGCUGAUG
2941
3369
CAUCAGCAUCGAAUCAGCC
3168

3369
GCAGGGAUCUAGGGGCCUC
2942
3369
GCAGGGAUCUAGGGGCCUC
2942
3387
GAGGCCCCUAGAUCCCUGC
3169

3387
CACUGAUCAGGCCAUAUUU
2943
3387
CACUGAUCAGGCCAUAUUU
2943
3405
AAAUAUGGCCUGAUCAGUG
3170

3405
UUAUGCUGUGACACCACUG
2944
3405
UUAUGCUGUGACACCACUG
2944
3423
CAGUGGUGUCACAGCAUAA
3171

3423
GCCCUGGUGUCCCCAUUUG
2945
3423
GCCCUGGUGUCCCCAUUUG
2945
3441
CAAAUGGGGACACCAGGGC
3172

3441
GGUGGCAGUAUGCCCCAUA
2946
3441
GGUGGCAGUAUGCCCCAUA
2946
3459
UAUGGGGCAUACUGCCACC
3173

3459
ACCUGCAGCAGGCCUAGAC
2947
3459
ACCUGCAGCAGGCCUAGAC
2947
3477
GUCUAGGCCUGCUGCAGGU
3174

3477
CGUGACCCAACCUUGUGGG
2948
3477
CGUGACCCAACCUUGUGGG
2948
3495
CCCACAAGGUUGGGUCACG
3175

3495
GGACUGUGGAACAAUCCAA
2949
3495
GGACUGUGGAACAAUCCAA
2949
3513
UUGGAUUGUUCCACAGUCC
3176

3513
AGAGAAUUGGGUGUGUCUC
2950
3513
AGAGAAUUGGGUGUGUCUC
2950
3531
GAGACACACCCAAUUCUCU
3177

3531
CUCUUGCUAUCAGGUCUAC
2951
3531
CUCUUGCUAUCAGGUCUAC
2951
3549
GUAGACCUGAUAGCAAGAG
3178

3549
CUGUGGUCGUUACAUCAAU
2952
3549
CUGUGGUCGUUACAUCAAU
2952
3567
AUUGAUGUAACGACCACAG
3179

3567
UGGCCACAUGCUCCAACAC
2953
3567
UGGCCACAUGCUCCAACAC
2953
3585
GUGUUGGAGCAUGUGGCCA
3180

3585
CCAUGGAAAUUCUGGACAC
2954
3585
CCAUGGAAAUUCUGGACAC
2954
3603
GUGUCCAGAAUUUCCAUGG
3181

3603
CCCGCUGGUCCUCAGCUAC
2955
3603
CCCGCUGGUCCUCAGCUAC
2955
3621
GUAGCUGAGGACCAGCGGG
3182

3621
CAUCGACCUGUCAGCCUGG
2956
3621
CAUCGACCUGUCAGCCUGG
2956
3639
CCAGGCUGACAGGUCGAUG
3183

3639
GUGUUACUACUGUCAGGCC
2957
3639
GUGUUACUACUGUCAGGCC
2957
3657
GGCCUGACAGUAGUAACAC
3184

3657
CUAUGUCCACCACCAGGCU
2958
3657
CUAUGUCCACCACCAGGCU
2958
3675
AGCCUGGUGGUGGACAUAG
3185

3675
UCUCCUAGAUGUGAAGAAC
2959
3675
UCUCCUAGAUGUGAAGAAC
2959
3693
GUUCUUCACAUCUAGGAGA
3186

3693
CAUCGCCCACCAGAACAAG
2960
3693
CAUCGCCCACCAGAACAAG
2960
3711
CUUGUUCUGGUGGGCGAUG
3187

3711
GUUUGGGGAGGAUAUGCCC
2961
3711
GUUUGGGGAGGAUAUGCCC
2961
3729
GGGCAUAUCCUCCCCAAAC
3188

3729
GCACCCACACUAAGCCCCA
2962
3729
CCACCCACACUAAGCCCCA
2962
3747
UGGGGCUUAGUGUGGGUGG
3189

3747
AGAAUACGGUCCCUCUUCA
2963
3747
AGAAUACGGUCCCUCUUCA
2963
3765
UGAAGAGGGACCGUAUUCU
3190

3765
ACCUUCUGAGGCCCACGAU
2964
3765
ACCUUCUGAGGCCCACGAU
2964
3783
AUCGUGGGCCUCAGAAGGU
3191

3783
UAGACCAGCUGUAGCUCAU
2965
3783
UAGACCAGCUGUAGCUCAU
2965
3801
AUGAGCUACAGCUGGUCUA
3192

3801
UUCCAGCCUGUACCUUGGA
2966
3801
UUCCAGCCUGUACCUUGGA
2966
3819
UCCAAGGUACAGGCUGGAA
3193

3819
AUGAGGGGUAGCCUCCCAC
2967
3819
AUGAGGGGUAGCCUCCCAC
2967
3837
GUGGGAGGCUACCCCUCAU
3194

3837
CUGCAUCCCAUCCUGAAUA
2968
3837
CUGCAUCCCAUCCUGAAUA
2968
3855
UAUUCAGGAUGGGAUGCAG
3195

3855
AUCCUUUGCAACUCCCCAA
2969
3855
AUCCUUUGCAACUCCCCAA
2969
3873
UUGGGGAGUUGCAAAGGAU
3196

3873
AGAGUGCUUAUUUAAGUGU
2970
3873
AGAGUGCUUAUUUAAGUGU
2970
3891
ACACUUAAAUAAGCACUCU
3197

3891
UUAAUACUUUUAAGAGAAC
2971
3891
UUAAUACUUUUAAGAGAAC
2971
3909
GUUCUCUUAAAAGUAUUAA
3198

3909
CUGCGACGAUUAAUUGUGG
2972
3909
CUGCGACGAUUAAUUGUGG
2972
3927
CCACAAUUAAUCGUCGCAG
3199

3927
GAUCUCCCCCUGCCCAUUG
2973
3927
GAUCUCCCCCUGCCCAUUG
2973
3945
CAAUGGGCAGGGGGAGAUC
3200

3945
GCCUGCUUGAGGGGCACCA
2974
3945
GCCUGCUUGAGGGGCACCA
2974
3963
UGGUGCCCCUCAAGCAGGC
3201

3963
ACUACUCCAGCCCAGAAGG
2975
3963
ACUACUCCAGCCCAGAAGG
2975
3981
GCUUCUGGGCUGGAGUAGU
3202

3981
GAAAGGGGGGCAGCUCAGU
2976
3981
GAAAGGGGGGCAGCUCAGU
2976
3999
ACUGAGCUGCCCCCCUUUC
3203

3999
UGGCCCCAAGAGGGAGCUG
2977
3999
UGGCCCCAAGAGGGAGCUG
2977
4017
CAGCUCCCUCUUGGGGCCA
3204

4017
GAUAUCAUGAGGAUAACAU
2978
4017
GAUAUCAUGAGGAUAACAU
2978
4035
AUGUUAUCCUCAUGAUAUC
3205

4035
UUGGCGGGAGGGGAGUUAA
2979
4035
UUGGCGGGAGGGGAGUUAA
2979
4053
UUAACUCCCCUCCCGCCAA
3206

4053
ACUGGCAGGCAUGGCAAGG
2980
4053
ACUGGCAGGCAUGGCAAGG
2980
4071
CCUUGCCAUGCCUGCCAGU
3207

4071
GUUGCAUAUGUAAUAAAGU
2981
4071
GUUGCAUAUGUAAUAAAGU
2981
4089
ACUUUAUUACAUAUGCAAC
3208

HDAC7: AF239243.1

3
AAUACCUACCUUGCAGGAC
3321
3
AAUACCUACCUUGCAGGAC
3321
21
GUCCUGCAAGGUAGGUAUU
3494

21
CCACGACAGGAUUAAGUGA
3322
21
CCACGACAGGAUUAAGUGA
3322
39
UCACUUAAUCCUGUCGUGG
3495

39
AGGAAAAACCCCCAUGAGA
3323
39
AGGAAAAACCCCCAUGAGA
3323
57
UCUCAUGGGGGUUUUUCCU
3496

57
AGUGUUUUGCCAUUGUCAA
3324
57
AGUGUUUUGCCAUUGUCAA
3324
75
UUGACAAUGGCAAAACACU
3497

75
AGUGAGCCUGAGGGAGGCU
3325
75
AGUGAGCCUGAGGGAGGCU
3325
93
AGCCUCCCUCAGGCUCACU
3498

93
UGAGGGGGGAUCAGGCUGU
3326
93
UGAGGGGGGAUCAGGCUGU
3326
111
ACAGCCUGAUCCCCCCUCA
3499

111
UAUCAUGCCCCCGAGGACA
3327
111
UAUCAUGCCCCCGAGGACA
3327
129
UGUCCUCGGGGGCAUGAUA
3500

129
AAACUUUCCAGUUUACCCU
3328
129
AAACUUUCCAGUUUACCCU
3328
147
AGGGUAAACUGGAAAGUUU
3501

147
UGCUCCCUCUCUCUGUCCC
3329
147
UGCUCCCUCUCUCUGUCCC
3329
165
GGGACAGAGAGAGGGAGCA
3502

165
CUAGGCUGCCCCAGGCCCU
3330
165
CUAGGCUGCCCCAGGCCCU
3330
183
AGGGCCUGGGGCAGCCUAG
3503

183
UGUGCAGACACACCAGGCC
3331
183
UGUGGAGACACACCAGGCC
3331
201
GGCCUGGUGUGUCUGCACA
3504

201
CCUCAGCCGCAGCCCAUGG
3332
201
CCUCAGCCGCAGCCCAUGG
3332
219
CCAUGGGCUGCGGCUGAGG
3505

219
GACCUGCGGGUGGGCCAGC
3333
219
GACCUGCGGGUGGGCCAGC
3333
237
GCUGGCCCACCCGCAGGUC
3506

237
CGGCCCCCAGUGGAGCCCC
3334
237
CGGCCCCCAGUGGAGCCCC
3334
255
GGGGCUCCACUGGGGGCCG
3507

255
CCACCAGAGCCCACAUUGC
3335
255
CCACCAGAGCCCACAUUGC
3335
273
GCAAUGUGGGCUCUGGUGG
3508

273
CUGGCCCUGCAGCGUCCCC
3336
273
CUGGCCCUGCAGCGUCCCC
3336
291
GGGGACGCUGCAGGGCCAG
3509

291
CAGCGCCUGCACCACCACC
3337
291
CAGCGCCUGCACCACCACC
3337
309
GGUGGUGGUGCAGGCGCUG
3510

309
CUCUUCCUAGCAGGCCUGC
3338
309
CUCUUCCUAGCAGGCCUGC
3338
327
GCAGGCCUGCUAGGAAGAG
3511

327
CAGCAGCAGCGCUCGGUGG
3339
327
CAGCAGCAGCGCUCGGUGG
3339
345
CCACCGAGCGCUGCUGCUG
3512

345
GAGCGCAUGAGGCUCUCCA
3340
345
GAGCCCAUGAGGCUCUCCA
3340
363
UGGAGAGCCUCAUGGGCUC
3513

363
AUGGACACGCCGAUGCCCG
3341
363
AUGGACACGCCGAUGCCCG
3341
381
CGGGCAUGGGCGUGUCCAU
3514

381
GAGUUGCAGGUGGGACCCC
3342
381
GAGUUGCAGGUGGGACCCC
3342
399
GGGGUCCCACCUGCAACUC
3515

399
CAGGAACAAGAGCUGCGGC
3343
399
CAGGAACAAGAGCUGCGGC
3343
417
GCCGCAGCUCUUGUUCCUG
3516

417
CAGCUUCUCCACAAGGACA
3344
417
CAGCUUCUCCACAAGGACA
3344
435
UGUCCUUGUGGAGAAGCUG
3517

435
AAGAGCAAGCGAAGUGCUG
3345
435
AAGAGCAAGCGAAGUGCUG
3345
453
CAGCACUUCGCUUGCUCUU
3518

453
GUAGCCAGCAGCGUGGUCA
3346
453
GUAGGCAGGAGCGUGGUCA
3346
471
UGACCACGCUGCUGGCUAC
3519

471
AAGCAGAAGCUAGCGGAGG
3347
471
AAGCAGAAGCUAGCGGAGG
3347
489
CCUCCGCUAGCUUCUGCUU
3520

489
GUGAUUCUGAAAAAACAGC
3348
489
GUGAUUCUGAAAAAACAGC
3348
507
GCUGUUUUUUCAGAAUCAC
3521

507
CAGGCGGCCCUAGAAAGAA
3349
507
CAGGCGGCCCUAGAAAGAA
3349
525
UUCUUUCUAGGGCCGCCUG
3522

525
ACAGUCCAUCCCAACAGCC
3350
525
ACAGUCCAUCCCAACAGCC
3350
543
GGCUGUUGGGAUGGACUGU
3523

543
CCCGGCAUUCCCUACAGAA
3351
543
CCCGGCAUUCCCUACAGAA
3351
561
UUCUGUAGGGAAUGCCGGG
3524

561
ACCCUGGAGCCCCUGGAGA
3352
561
ACCCUGGAGCCCCUGGAGA
3352
579
UCUCCAGGGGCUCCAGGGU
3525

579
ACGGAAGGAGCCACCCGCU
3353
579
ACGGAAGGAGCCACCCGCU
3353
597
AGCGGGUGGCUCCUUCCGU
3526

597
UCCAUGCUCAGCAGCUUUU
3354
597
UCCAUGCUCAGCAGCUUUU
3354
615
AAAAGCUGCUGAGCAUGGA
3527

615
UUGCCUCCUGUUCCCAGCC
3355
615
UUGCCUCCUGUUCCCAGCC
3355
633
GGCUGGGAACAGGAGGCAA
3528

633
CUGCCCAGUGACCCCCCAG
3356
633
CUGCCCAGUGACCCCCCAG
3356
651
CUGGGGGGUCACUGGGCAG
3529

651
GAGCACUUCCCUCUGCGCA
3357
651
GAGCACUUCCCUCUGCGCA
3357
669
UGCGCAGAGGGAAGUGCUC
3530

669
AAGACAGUCUCUGAGCCCA
3358
669
AAGACAGUCUCUGAGCCCA
3358
687
UGGGCUCAGAGACUGUCUU
3531

687
AACCUGAAGCUGCGCUAUA
3359
687
AACCUGAAGCUGCGCUAUA
3359
705
UAUAGCGCAGCUUCAGGUU
3532

705
AAGCCCAAGAAGUCCCUGG
3360
705
AAGCCCAAGAAGUCCCUGG
3360
723
CCAGGGACUUCUUGGGCUU
3533

723
GAGCGGAGGAAGAAUCCAC
3361
723
GAGCGGAGGAAGAAUCCAC
3361
741
GUGGAUUCUUCCUCCGCUC
3534

741
CUGCUCCGAAAGGAGAGUG
3362
741
CUGCUCCGAAAGGAGAGUG
3362
759
CACUCUCCUUUCGGAGCAG
3535

759
GCGCCCCCCAGCCUCCGGC
3363
759
GCGCCCCCCAGCCUCCGGC
3363
777
GCCGGAGGCUGGGGGGCGC
3536

777
CGGCGGCCCGCAGAGACCC
3364
777
CGGCGGCCCGCAGAGACCC
3364
795
GGGUCUCUGCGGGCCGCCG
3537

795
CUCGGAGACUCCUCCCCAA
3365
795
CUCGGAGACUCCUCCCCAA
3365
813
UUGGGGAGGAGUCUCCGAG
3538

813
AGUAGUAGCAGCACGCCCG
3366
813
AGUAGUAGCAGCACGCCCG
3366
831
CGGGCGUGCUGCUACUACU
3539

831
GCAUCAGGGUGCAGCUCCC
3367
831
GCAUCAGGGUGCAGCUCCC
3367
849
GGGAGCUGCACCCUGAUGC
3540

849
CCCAAUGACAGCGAGCACG
3368
849
CCCAAUGACAGCGAGCACG
3368
867
CGUGCUCGCUGUCAUUGGG
3541

867
GGCCCCAAUCCCAUCCUGG
3369
867
GGCCCCAAUCCCAUCCUGG
3369
885
CCAGGAUGGGAUUGGGGCC
3542

885
GGCGACAGUGACCGCAGGA
3370
885
GGCGACAGUGACCGCAGGA
3370
903
UCCUGCGGUCACUGUCGCC
3543

903
ACCCAUCCGACUCUGGGCC
3371
903
ACCCAUCCGACUCUGGGCC
3371
921
GGCCCAGAGUCGGAUGGGU
3544

921
CCUCGGGGGCCAAUCCUGG
3372
921
CCUCGGGGGCCAAUCCUGG
3372
939
CCAGGAUUGGCCCCCGAGG
3545

939
GGGAGCCCCCACACUCCCC
3373
939
GGGAGCCCCCACACUCCCC
3373
957
GGGGAGUGUGGGGGCUCCC
3546

957
CUCUUCCUGCCCCAUGGCU
3374
957
CUCUUCCUGCCCCAUGGCU
3374
975
AGCCAUGGGGCAGGAAGAG
3547

975
UUGGAGCCCGAGGCUGGGG
3375
975
UUGGAGCCCGAGGCUGGGG
3375
993
CCCCAGCCUCGGGCUCCAA
3548

993
GGCACCUUGCCCUCUCGCC
3376
993
GGCACCUUGCCCUCUCGCC
3376
1011
GGCGAGAGGGCAAGGUGCC
3549

1011
CUGCAGCCCAUUCUCCUCC
3377
1011
CUGCAGCCCAUUCUCCUCC
3377
1029
GGAGGAGAAUGGGCUGCAG
3550

1029
CUGGACCCCUCAGGCUCUC
3378
1029
CUGGACCCCUCAGGCUCUC
3378
1047
GAGAGCCUGAGGGGUCCAG
3551

1047
CAUGCCCCGCUGCUGACUG
3379
1047
CAUGCCCCGCUGGUGACUG
3379
1065
CAGUCAGCAGCGGGGCAUG
3552

1065
GUGCCCGGGCUUGGGCCCU
3380
1065
GUGCCCGGGCUUGGGCCCU
3380
1083
AGGGCCCAAGCCCGGGCAC
3553

1083
UUGCCCUUCCACUUUGCCC
3381
1083
UUGCCCUUCCACUUUGCCC
3381
1101
GGGCAAAGUGGAAGGGCAA
3554

1101
CAGUCCUUAAUGACCACCG
3382
1101
CAGUCCUUAAUGACCACCG
3382
1119
CGGUGGUCAUUAAGGACUG
3555

1119
GAGCGGCUCUCUGGGUCAG
3383
1119
GAGCGGCUCUCUGGGUCAG
3383
1137
CUGACCCAGAGAGCCGCUC
3556

1137
GGCCUCCACUGGCCACUGA
3384
1137
GGCCUCCACUGGCCACUGA
3384
1155
UCAGUGGCCAGUGGAGGCC
3557

1155
AGCCGGACUGGCUCAGAGC
3385
1155
AGCCGGACUCGCUCAGAGC
3385
1173
GCUCUGAGCGAGUCCGGCU
3558

1173
CCCCUGCCCCCCAGUGCCA
3386
1173
CCCCUGCCCCCCAGUGCCA
3386
1191
UGGCACUGGGGGGGAGGGG
3559

1191
ACCGCUCCCCCAGCGCCGG
3387
1191
ACCGCUCCCCCACCGCCGG
3387
1209
CCGGCGGUGGGGGAGCGGU
3560

1209
GGCCCCAUGCAGCCCCGCC
3388
1209
GGCCCCAUGCAGCCCCGCC
3388
1227
GGCGGGGCUGCAUGGGGCC
3561

1227
CUGGAGCAGCUCAAAACUC
3389
1227
CUGGAGCAGCUCAAAACUC
3389
1245
GAGUUUUGAGCUGCUCCAG
3562

1245
CACGUCCAGGUGAUCAAGA
3390
1245
CACGUCCAGGUGAUCAAGA
3390
1263
UCUUGAUCACCUGGACGUG
3563

1263
AGGUCAGCCAAGCCGAGUG
3391
1263
AGGUCAGCCAAGCCGAGUG
3391
1281
CACUCGGCUUGGCUGACCU
3564

1281
GAGAAGCCCCGGCUGCGGC
3392
1281
GAGAAGCCCCGGCUGCGGC
3392
1299
GCCGCAGCCGGGGCUUCUC
3565

1299
CAGAUACCCUCGGCUGAAG
3393
1299
CAGAUACCCUCGGCUGAAG
3393
1317
CUUCAGCCGAGGGUAUCUG
3566

1317
GACCUGGAGACAGAUGGCG
3394
1317
GACCUGGAGACAGAUGGCG
3394
1335
CGCCAUCUGUCUCCAGGUC
3567

1335
GGGGGACCGGGCCAGGUGG
3395
1335
GGGGGACCGGGCCAGGUGG
3395
1353
CCACCUGGCCCGGUCCCCC
3568

1353
GUGGACGAUGGCCUGGAGC
3396
1353
GUGGACGAUGGCCUGGAGC
3396
1371
GCUCCAGGCCAUCGUCCAC
3569

1371
CACAGGGAGCUGGGCCAUG
3397
1371
CACAGGGAGCUGGGCCAUG
3397
1389
CAUGGCCCAGCUCCCUGUG
3570

1389
GGGCAGCCUGAGGCCAGAG
3398
1389
GGGCAGCCUGAGGCCAGAG
3398
1407
CUCUGGCCUCAGGCUGCCC
3571

1407
GGCCCCGCUCCUCUCCAGC
3399
1407
GGCCCCGCUCCUCUCCAGC
3399
1425
GCUGGAGAGGAGCGGGGCC
3572

1425
GAGCACCCUCAGGUGUUGC
3400
1425
CAGCACCCUCAGGUGUUGC
3400
1443
GCAACACCUGAGGGUGCUG
3573

1443
CUCUGGGAACAGCAGCGAC
3401
1443
CUCUGGGAACAGCAGCGAC
3401
1461
GUCGCUGCUGUUCCCAGAG
3574

1461
CUGGCUGGGCGGCUCCCCC
3402
1461
CUGGCUGGGCGGCUCCCCC
3402
1479
GGGGGAGCCGCCCAGCCAG
3575

1479
CGGGGCAGCACCGGGGACA
3403
1479
CGGGGCAGCACCGGGGACA
3403
1497
UGUCCCCGGUGCUGCCCCG
3576

1497
ACUGUGCUGCUUCCUCUGG
3404
1497
ACUGUGCUGCUUCCUCUGG
3404
1515
CCAGAGGAAGCAGCACAGU
3577

1515
GCCCAGGGUGGGCACCGGC
3405
1515
GCCCAGGGUGGGCACCGGC
3405
1533
GCCGGUGCCCACCCUGGGC
3578

1533
CCUCUGUCCCGGGCUCAGU
3406
1533
CCUCUGUCCCGGGCUCAGU
3406
1551
ACUGAGCCCGGGACAGAGG
3579

1551
UCUUCCCCAGCCGCACCUG
3407
1551
UCUUCCCCAGCCGCACCUG
3407
1569
CAGGUGCGGCUGGGGAAGA
3580

1569
GCCUCACUGUCAGCCCCAG
3408
1569
GCCUCACUGUCAGCCCCAG
3408
1587
CUGGGGCUGACAGUGAGGC
3581

1587
GAGCCUGCCAGCCAGGCCC
3409
1587
GAGCCUGCCAGCCAGGCCC
3409
1605
GGGCCUGGCUGGCAGGCUC
3582

1605
CGAGUCCUCUCCAGCUCAG
3410
1605
GGAGUCCUCUCCAGCUCAG
3410
1623
CUGAGCUGGAGAGGACUCG
3583

1623
GAGACCCCUGCCAGGACCC
3411
1623
GAGACCCCUGCCAGGACCC
3411
1641
GGGUCCUGGCAGGGGUCUC
3584

1641
CUGCCCUUCACCACAGGGC
3412
1641
CUGCCCUUCACCACAGGGC
3412
1659
GCCCUGUGGUGAAGGGCAG
3585

1659
CUGAUCUAUGACUCGGUCA
3413
1659
CUGAUCUAUGACUCGGUCA
3413
1677
UGACCGAGUCAUAGAUCAG
3586

1677
AUGCUGAAGCACCAGUGCU
3414
1677
AUGCUGAAGCACCAGUGCU
3414
1695
AGCACUGGUGCUUCAGCAU
3587

1695
UCCUGCGGUGACAACAGCA
3415
1695
UCCUGCGGUGACAACAGCA
3415
1713
UGCUGUUGUCACCGCAGGA
3588

1713
AGGCACCCGGAGCACGCCG
3416
1713
AGGCACCCGGAGCACGCCG
3416
1731
CGGCGUGCUCCGGGUGCCU
3589

1731
GGCCGCAUCCAGAGCAUCU
3417
1731
GGCCGCAUCCAGAGCAUCU
3417
1749
AGAUGCUCUGGAUGCGGCC
3590

1749
UGGUCCCGGCUGCAGGAGC
3418
1749
UGGUCCCGGCUGCAGGAGC
3418
1767
GCUCCUGCAGCCGGGACCA
3591

1767
CGGGGGCUCCGGAGCCAGU
3419
1767
CGGGGGCUCCGGAGCCAGU
3419
1785
ACUGGCUCCGGAGCCCCCG
3592

1785
UGUGAGUGUCUCCGAGGCC
3420
1785
UGUGAGUGUCUCCGAGGCC
3420
1803
GGCCUCGGAGACACUCACA
3593

1803
CGGAAGGCCUCCCUGGAAG
3421
1803
CGGAAGGCCUCCCUGGAAG
3421
1821
CUUCCAGGGAGGCCUUCCG
3594

1821
GAGCUGCAGUCGGUCCACU
3422
1821
GAGCUGCAGUCGGUCCACU
3422
1839
AGUGGACCGACUGCAGCUC
3595

1839
UCUGAGCGGCACGUGCUCC
3423
1839
UCUGAGCGGCACGUGCUCC
3423
1857
GGAGCACGUGCCGCUCAGA
3596

1857
CUCUACGGCACCAACCCGC
3424
1857
CUCUACGGCACCAACCCGC
3424
1875
GCGGGUUGGUGCCGUAGAG
3597

1875
CUCAGCCGCCUCAAACUGG
3425
1875
CUCAGCCGCCUCAAACUGG
3425
1893
CCAGUUUGAGGCGGCUGAG
3598

1893
GACAACGGGAAGCUGGCAG
3426
1893
GACAACGGGAAGCUGGCAG
3426
1911
CUGCCAGCUUCCCGUUGUC
3599

1911
GGGCUCCUGGCACAGCGGA
3427
1911
GGGCUCCUGGCACAGCGGA
3427
1929
UCCGCUGUGCCAGGAGCCC
3600

1929
AUGUUUGAGAUGCUGCCCU
3428
1929
AUGUUUGAGAUGCUGCCCU
3428
1947
AGGGCAGCAUCUCAAACAU
3601

1947
UGUGGUGGGGUUGGGGUGG
3429
1947
UGUGGUGGGGUUGGGGUGG
3429
1965
CCACCCCAACCCCACCACA
3602

1965
GACACUGACACCAUCUGGA
3430
1965
GACACUGACACCAUCUGGA
3430
1983
UCCAGAUGGUGUCAGUGUC
3603

1983
AAUGAGCUUCAUUCCUCCA
3431
1983
AAUGAGCUUCAUUCCUCCA
3431
2001
UGGAGGAAUGAAGCUCAUU
3604

2001
AAUGCAGCCCGCUGGGCCG
3432
2001
AAUGCAGCCCGCUGGGCCG
3432
2019
CGGCCCAGCGGGCUGCAUU
3605

2019
GCUGGCAGUGUCACUGACC
3433
2019
GCUGGCAGUGUCACUGACC
3433
2037
GGUCAGUGACACUGCCAGC
3606

2037
CUCGCCUUCAAAGUGGCUU
3434
2037
CUCGCCUUCAAAGUGGCUU
3434
2055
AAGCCACUUUGAAGGCGAG
3607

2055
UCUCGUGAGCUAAAGAAUG
3435
2055
UCUCGUGAGCUAAAGAAUG
3435
2073
CAUUCUUUAGCUCACGAGA
3608

2073
GGUUUCGCUGUGGUGCGGC
3436
2073
GGUUUCGCUGUGGUGCGGC
3436
2091
GCCGCACCACAGCGAAACC
3609

2091
CCCCCAGGACACCAUGCAG
3437
2091
CCCCCAGGACACCAUGCAG
3437
2109
CUGCAUGGUGUCCUGGGGG
3610

2109
GAUCAUUCAACAGCCAUGG
3438
2109
GAUCAUUCAACAGCCAUGG
3438
2127
CCAUGGCUGUUGAAUGAUC
3611

2127
GGCUUCUGCUUCUUCAACU
3439
2127
GGCUUCUGCUUCUUCAACU
3439
2145
AGUUGAAGAAGCAGAAGCC
3612

2145
UCAGUGGCCAUCGCCUGCC
3440
2145
UCAGUGGCCAUCGCCUGCC
3440
2163
GGCAGGCGAUGGCCACUGA
3613

2163
CGGCAGCUGCAACAGCAGA
3441
2163
CGGCAGCUGCAACAGCAGA
3441
2181
UCUGCUGUUGCAGCUGCCG
3614

2181
AGCAAGGCCAGCAAGGCCA
3442
2181
AGCAAGGCCAGCAAGGCCA
3442
2199
UGGCCUUGCUGGCCUUGCU
3615

2199
AGCAAGAUCCUCAUUGUAG
3443
2199
AGCAAGAUCCUCAUUGUAG
3443
2217
CUACAAUGAGGAUCUUGCU
3616

2217
GACUGGGACGUGCACCAUG
3444
2217
GACUGGGACGUGCACCAUG
3444
2235
CAUGGUGCACGUCCCAGUC
3617

2235
GGCAACGGCACCCAGCAAA
3445
2235
GGCAACGGCACCCAGCAAA
3445
2253
UUUGCUGGGUGCCGUUGCC
3618

2253
ACCUUCUACCAAGACCCCA
3446
2253
ACCUUCUACCAAGACCCCA
3446
2271
UGGGGUCUUGGUAGAAGGU
3619

2271
AGUGUGCUCUACAUCUCCC
3447
2271
AGUGUGCUCUACAUCUCCC
3447
2289
GGGAGAUGUAGAGCACACU
3620

2289
CUGCAUCGCCAUGACGACG
3448
2289
CUGCAUCGCCAUGACGACG
3448
2307
CGUCGUCAUGGCGAUGCAG
3621

2307
GGCAACUUCUUCCCGGGGA
3449
2307
GGCAACUUCUUCCCGGGGA
3449
2325
UCCCCGGGAAGAAGUUGCC
3622

2325
AGUGGGGCUGUGGAUGAGG
3450
2325
AGUGGGGCUGUGGAUGAGG
3450
2343
CCUCAUCCACAGCCCCACU
3623

2343
GUAGGGGCUGGCAGCGGUG
3451
2343
GUAGGGGCUGGCAGCGGUG
3451
2361
CACCGCUGCCAGCCCCUAC
3624

2361
GAGGGCUUCAAUGUCAAUG
3452
2361
GAGGGCUUCAAUGUCAAUG
3452
2379
CAUUGACAUUGAAGCCCUC
3625

2379
GUGGCCUGGGCUGGAGGUC
3453
2379
GUGGCCUGGGCUGGAGGUC
3453
2397
GACCUCCAGCCCAGGCCAC
3626

2397
CUGGACCCCCCCAUGGGGG
3454
2397
CUGGACCCCCCCAUGGGGG
3454
2415
CCCCCAUGGGGGGGUCCAG
3627

2415
GAUCCUGAGUACCUGGCUG
3455
2415
GAUCCUGAGUACCUGGCUG
3455
2433
CAGCCAGGUACUCAGGAUC
3628

2433
GCUUUCAGGAUAGUCGUGA
3456
2433
GCUUUCAGGAUAGUCGUGA
3456
2451
UCACGACUAUCCUGAAAGC
3629

2451
AUGCCCAUCGCCCGAGAGU
3457
2451
AUGCCCAUCGCCCGAGAGU
3457
2469
ACUCUCGGGCGAUGGGCAU
3630

2469
UUCUCUCCAGACCUAGUCC
3458
2469
UUCUCUCCAGACCUAGUCC
3458
2487
GGACUAGGUCUGGAGAGAA
3631

2487
CUGGUGUCUGCUGGAUUUG
3459
2487
CUGGUGUCUGCUGGAUUUG
3459
2505
CAAAUCCAGCAGACACCAG
3632

2505
GAUGCUGCUGAGGGUCACC
3460
2505
GAUGCUGCUGAGGGUCACC
3460
2523
GGUGACCCUCAGCAGCAUC
3633

2523
CCGGCCCCACUGGGUGGCU
3461
2523
CCGGCCCCACUGGGUGGCU
3461
2541
AGCCACCCAGUGGGGCCGG
3634

2541
UACCAUGUUUCUGCCAAAU
3462
2541
UACCAUGUUUCUGCCAAAU
3462
2559
AUUUGGCAGAAACAUGGUA
3635

2559
UGUUUUGGAUACAUGACGC
3463
2559
UGUUUUGGAUACAUGACGC
3463
2577
GCGUCAUGUAUCCAAAACA
3636

2577
CAGCAACUGAUGAACCUGG
3464
2577
CAGCAACUGAUGAACCUGG
3464
2595
CCAGGUUCAUCAGUUGCUG
3637

2595
GCAGGAGGCGCAGUGGUGC
3465
2595
GCAGGAGGCGCAGUGGUGC
3465
2613
GCACCACUGCGCCUCCUGC
3638

2613
CUGGCCUUGGAGGGUGGCC
3466
2613
CUGGCCUUGGAGGGUGGCC
3466
2631
GGCCACCCUCCAAGGCCAG
3639

2631
CAUGACCUCACAGCCAUCU
3467
2631
CAUGACCUCACAGCCAUCU
3467
2649
AGAUGGCUGUGAGGUCAUG
3640

2649
UGUGACGCCUCUGAGGCCU
3468
2649
UGUGACGCCUCUGAGGCCU
3468
2667
AGGCCUCAGAGGCGUCACA
3641

2667
UGUGUGGCUGCUCUUCUGG
3469
2667
UGUGUGGCUGCUCUUCUGG
3469
2685
CCAGAAGAGCAGCCACACA
3642

2685
GGUAACAGGGUGGAUCCCC
3470
2685
GGUAACAGGGUGGAUCCCC
3470
2703
GGGGAUCCACCCUGUUACC
3643

2703
CUUUCAGAAGAAGGCUGGA
3471
2703
CUUUCAGAAGAAGGCUGGA
3471
2721
UCCAGCCUUCUUCUGAAAG
3644

2721
AAACAGAAACCCCAACCUC
3472
2721
AAACAGAAACCCCAACCUC
3472
2739
GAGGUUGGGGUUUCUGUUU
3645

2739
CAAUGCCAUCCGCUCUCUG
3473
2739
CAAUGCCAUCCGCUCUCUG
3473
2757
CAGAGAGCGGAUGGCAUUG
3646

2757
GGAGGCCGUGAUCCGGGUG
3474
2757
GGAGGCCGUGAUCCGGGUG
3474
2775
CACCCGGAUCACGGCCUCC
3647

2775
GCACAGUAAAUACUGGGGC
3475
2775
GCACAGUAAAUACUGGGGC
3475
2793
GCCCCAGUAUUUACUGUGC
3648

2793
CUGCAUGCAGCGCCUGGCC
3476
2793
CUGCAUGCAGCGCCUGGCC
3476
2811
GGCCAGGCGCUGCAUGCAG
3649

2811
CUCCUGUCCAGACUCCUGG
3477
2811
CUCCUGUCCAGACUCCUGG
3477
2829
CCAGGAGUCUGGACAGGAG
3650

2829
GGUGCCUAGAGUGCCAGGG
3478
2829
GGUGCCUAGAGUGCCAGGG
3478
2847
CCCUGGCACUCUAGGCACC
3651

2847
GGCUGACAAAGAAGAAGUG
3479
2847
GGCUGACAAAGAAGAAGUG
3479
2865
CACUUCUUCUUUGUCAGCC
3652

2865
GGAGGCAGUGACCGCACUG
3480
2865
GGAGGCAGUGACCGCACUG
3480
2883
CAGUGCGGUCACUGCCUCC
3653

2883
GGCGUCCCUCUCUGUGGGC
3481
2883
GGCGUCCCUCUCUGUGGGC
3481
2901
GCCCACAGAGAGGGACGCC
3654

2901
CAUCCUGGCUGAAGAUAGG
3482
2901
CAUCCUGGCUGAAGAUAGG
3482
2919
CCUAUCUUCAGCCAGGAUG
3655

2919
GCCCUCGGAGCAGCUGGUG
3483
2919
GCCCUCGGAGCAGCUGGUG
3483
2937
CACCAGCUGCUCCGAGGGC
3656

2937
GGAGGAGGAAGAACCUAUG
3484
2937
GGAGGAGGAAGAACCUAUG
3484
2955
CAUAGGUUCUUCCUCCUCC
3657

2955
GAAUCUCUAAGGCUCUGGA
3485
2955
GAAUCUCUAAGGCUCUGGA
3485
2973
UCCAGAGCCUUAGAGAUUC
3658

2973
AACCAUCUGCCCGCCCACC
3486
2973
AACCAUCUGCCCGCCCACC
3486
2991
GGUGGGCGGGCAGAUGGUU
3659

2991
CAUGCCCUUGGGACCUGGU
3487
2991
CAUGCCCUUGGGACCUGGU
3487
3009
ACCAGGUCCCAAGGGCAUG
3660

3009
UUCUCUUCUAACCCCUGGC
3488
3009
UUCUCUUCUAACCCCUGGC
3488
3027
GCCAGGGGUUAGAAGAGAA
3661

3027
CAAUAGCCCCCAUUCCUGG
3489
3027
CAAUAGCCCCCAUUCCUGG
3489
3045
CCAGGAAUGGGGGCUAUUG
3662

3045
GGUCUUUAGAGAUCCUGUG
3490
3045
GGUCUUUAGAGAUCCUGUG
3490
3063
CACAGGAUCUCUAAAGACC
3663

3063
GGGCAAGUAGUUGGAACCA
3491
3063
GGGCAAGUAGUUGGAACCA
3491
3081
UGGUUCCAACUACUUGCCC
3664

3081
AGAGAACAGCCUGCCUGCU
3492
3081
AGAGAACAGGGUGCCUGCU
3492
3099
AGCAGGCAGGCUGUUCUCU
3665

3099
UUUGACAGUUAUCCCAGGG
3493
3099
UUUGACAGUUAUCCCAGGG
3493
3117
CCCUGGGAUAACUGUCAAA
3666

HIDAC8: NM_018486.1

3
CAGAUCUGGAAGGUGGCUG
3779
3
CAGAUCUGGAAGGUGGCUG
3779
21
CAGGCACCUUCCAGAUCUG
3875

21
GCGGAACGGUUUUAAGCGG
3780
21
GCGGAACGGUUUUAAGCGG
3780
39
CCGCUUAAAACCGUUCCGC
3876

39
GAAGAUGGAGGAGCCGGAG
3781
39
GAAGAUGGAGGAGCCGGAG
3781
57
CUCCGGCUCCUCCAUCUUC
3877

57
GGAACCGGCGGACAGUGGG
3782
57
GGAACCGGCGGACAGUGGG
3782
75
CCCACUGUCCGCCGGUUCC
3878

75
GCAGUCGCUGGUCCCGGUU
3783
75
GCAGUCGCUGGUCCCGGUU
3783
93
AAGCGGGACCAGCGACUGC
3879

93
UUAUAUCUAUAGUCCCGAG
3784
93
UUAUAUCUAUAGUCCCGAG
3784
111
CUCGGGACUAUAGAUAUAA
3880

111
GUAUGUCAGUAUGUGUGAC
3785
111
GUAUGUCAGUAUGUGUGAC
3785
129
GUCACACAUACUGACAUAC
3881

129
CUCCCUGGCCAAGAUCCCC
3786
129
CUCCCUGGCCAAGAUCCCC
3786
147
GGGGAUCUUGGCCAGGGAG
3882

147
CAAACGGGCCAGUAUGGUG
3787
147
CAAACGGGCCAGUAUGGUG
3787
165
CACCAUACUGGCCCGUUUG
3883

165
GCAUUCUUUGAUUGAAGCA
3788
165
GCAUUCUUUGAUUGAAGCA
3788
183
UGCUUCAAUCAAAGAAUGC
3884

183
AUAUGCACUGCAUAAGCAG
3789
183
AUAUGCACUGCAUAAGCAG
3789
201
CUGCUUAUGCAGUGCAUAU
3885

201
GAUGAGGAUAGUUAAGCCU
3790
201
GAUGAGGAUAGUUAAGCCU
3790
219
AGGCUUAACUAUCCUCAUC
3886

219
UAAAGUGGCCUCCAUGGAG
3791
219
UAAAGUGGCCUCCAUGGAG
3791
237
CUCCAUGGAGGCCACUUUA
3887

237
GGAGAUGGCCACCUUCCAC
3792
237
GGAGAUGGCCACCUUCCAC
3792
255
GUGGAAGGUGGCCAUCUCC
3888

255
CACUGAUGCUUAUCUGCAG
3793
255
CACUGAUGCUUAUCUGCAG
3793
273
CUGCAGAUAAGCAUCAGUG
3889

273
GCAUCUCCAGAAGGUCAGC
3794
273
GCAUCUCCAGAAGGUCAGC
3794
291
GCUGACCUUCUGGAGAUGC
3890

291
CCAAGAGGGCGAUGAUGAU
3795
291
CCAAGAGGGCGAUGAUGAU
3795
309
AUCAUCAUCGCCCUCUUGG
3891

309
UCAUCCGGACUCCAUAGAA
3796
309
UCAUCCGGACUCCAUAGAA
3796
327
UUCUAUGGAGUCCGGAUGA
3892

327
AUAUGGGCUAGGUUAUGAC
3797
327
AUAUGGGCUAGGUUAUGAC
3797
345
GUCAUAACCUAGCCCAUAU
3893

345
CUGCCCAGCCACUGAAGGG
3798
345
CUGCCCAGCCACUGAAGGG
3798
363
CCCUUCAGUGGCUGGGCAG
3894

363
GAUAUUUGACUAUGCAGCA
3799
363
GAUAUUUGACUAUGCAGCA
3799
381
UGCUGCAUAGUCAAAUAUC
3895

381
AGCUAUAGGAGGGGCUACG
3800
381
AGCUAUAGGAGGGGCUACG
3800
399
CGUAGCCCCUCCUAUAGCU
3896

399
GAUCACAGCUGCCCAAUGC
3801
399
GAUCACAGCUGCCCAAUGC
3801
417
GCAUUGGGCAGCUGUGAUC
3897

417
CCUGAUUGACGGAAUGUGC
3802
417
CCUGAUUGACGGAAUGUGC
3802
435
GCACAUUCCGUCAAUCAGG
3898

435
CAAAGUAGCAAUUAACUGG
3803
435
CAAAGUAGCAAUUAACUGG
3803
453
CCAGUUAAUUGCUACUUUG
3899

453
GUCUGGAGGGUGGCAUCAU
3804
453
GUCUGGAGGGUGGCAUCAU
3804
471
AUGAUGCCACCCUCCAGAC
3900

471
UGCAAAGAAAGAUGAAGCA
3805
471
UGCAAAGAAAGAUGAAGCA
3805
489
UGCUUCAUCUUUCUUUGCA
3901

489
AUCUGGUUUUUGUUAUCUC
3806
489
AUCUGGUUUUUGUUAUCUC
3806
507
GAGAUAACAAAAACCAGAU
3902

507
CAAUGAUGCUGUCCUGGGA
3807
507
CAAUGAUGCUGUCCUGGGA
3807
525
UCCCAGGACAGCAUCAUUG
3903

525
AAUAUUACGAUUGCGACGG
3808
525
AAUAUUACGAUUGCGACGG
3808
543
CCGUCGCAAUCGUAAUAUU
3904

543
GAAAUUUGAGCGUAUUCUC
3809
543
GAAAUUUGAGCGUAUUCUC
3809
561
GAGAAUACGCUCAAAUUUC
3905

561
CUACGUGGAUUUGGAUCUG
3810
561
CUACGUGGAUUUGGAUCUG
3810
579
CAGAUCCAAAUCCACGUAG
3906

579
GCACCAUGGAGAUGGUGUA
3811
579
GCACCAUGGAGAUGGUGUA
3811
597
UACACCAUCUCCAUGGUGC
3907

597
AGAAGACGCAUUCAGUUUC
3812
597
AGAAGACGCAUUCAGUUUC
3812
615
GAAACUGAAUGCGUCUUCU
3908

615
CACCUCCAAAGUCAUGACC
3813
615
CACCUCCAAAGUCAUGACC
3813
633
GGUCAUGACUUUGGAGGUG
3909

633
CGUGUCCCUGCACAAAUUC
3814
633
CGUGUCCCUGCACAAAUUC
3814
651
GAAUUUGUGCAGGGACACG
3910

651
CUCCCCAGGAUUUUUCCCA
3815
651
CUCCCCAGGAUUUUUCCCA
3815
669
UGGGAAAAAUCCUGGGGAG
3911

669
AGGAACAGGUGACGUGUCU
3816
669
AGGAACAGGUGACGUGUCU
3816
687
AGACACGUCACCUGUUCCU
3912

687
UGAUGUUGGCCUAGGGAAG
3817
687
UGAUGUUGGCCUAGGGAAG
3817
705
CUUCCCUAGGCCAACAUCA
3913

705
GGGACGGUACUACAGUGUA
3818
705
GGGACGGUACUACAGUGUA
3818
723
UACACUGUAGUACCGUCCC
3914

723
AAAUGUGCCCAUUCAGGAU
3819
723
AAAUGUGCCCAUUCAGGAU
3819
741
AUCCUGAAUGGGCACAUUU
3915

741
UGGCAUACAAGAUGAAAAA
3820
741
UGGCAUACAAGAUGAAAAA
3820
759
UUUUUCAUCUUGUAUGCCA
3916

759
AUAUUACCAGAUCUGUGAA
3821
759
AUAUUACCAGAUCUGUGAA
3821
777
UUCACAGAUCUGGUAAUAU
3917

777
AAGUGUACUAAAGGAAGUA
3822
777
AAGUGUACUAAAGGAAGUA
3822
795
UACUUCCUUUAGUACACUU
3918

795
AUACCAAGCCUUUAAUCCC
3823
795
AUACCAAGCCUUUAAUCCC
3823
813
GGGAUUAAAGGCUUGGUAU
3919

813
CAAAGCAGUGGUCUUACAG
3824
813
CAAAGCAGUGGUCUUACAG
3824
831
CUGUAAGACCACUGCUUUG
3920

831
GCUGGGAGCUGACACAAUA
3825
831
GCUGGGAGCUGACACAAUA
3825
849
UAUUGUGUCAGCUCCCAGC
3921

849
AGCUGGGGAUCGCAUGUGC
3826
849
AGCUGGGGAUCCCAUGUGC
3826
867
GCACAUGGGAUCCCCAGCU
3922

867
CUCCUUUAACAUGACUCCA
3827
867
CUCCUUUAACAUGACUCCA
3827
885
UGGAGUCAUGUUAAAGGAG
3923

885
AGUGGGAAUUGGCAAGUGU
3828
885
AGUGGGAAUUGGCAAGUGU
3828
903
ACACUUGCCAAUUCCCACU
3924

903
UCUUAAGUACAUCCUUCAA
3829
903
UCUUAAGUACAUCCUUCAA
3829
921
UUGAAGGAUGUACUUAAGA
3925

921
AUGGCAGUUGGCAACACUC
3830
921
AUGGCAGUUGGCAACACUC
3830
939
GAGUGUUGCCAACUGCCAU
3926

939
CAUUUUGGGAGGAGGAGGC
3831
939
CAUUUUGGGAGGAGGAGGC
3831
957
GCCUCCUCCUCCCAAAAUG
3927

957
CUAUAACCUUGCCAACACG
3832
957
CUAUAACCUUGCCAACACG
3832
975
CGUGUUGGCAAGGUUAUAG
3928

975
GGCUCGAUGCUGGACAUAC
3833
975
GGCUCGAUGCUGGACAUAC
3833
993
GUAUGUCCAGCAUGGAGCC
3929

993
CUUGACCGGGGUCAUCCUA
3834
993
CUUGACCGGGGUCAUCCUA
3834
1011
UAGGAUGACCCCGGUCAAG
3930

1011
AGGGAAAACACUAUCCUCU
3835
1011
AGGGAAAACACUAUCCUCU
3835
1029
AGAGGAUAGUGUUUUCCCU
3931

1029
UGAGAUCCCAGAUCAUGAG
3836
1029
UGAGAUCCCAGAUCAUGAG
3836
1047
CUCAUGAUCUGGGAUCUCA
3932

1047
GUUUUUCACAGCAUAUGGU
3837
1047
GUUUUUCACAGCAUAUGGU
3837
1065
ACCAUAUGCUGUGAAAAAC
3933

1065
UCCUGAUUAUGUGCUGGAA
3838
1065
UCCUGAUUAUGUGCUGGAA
3838
1083
UUCCAGCACAUAAUCAGGA
3934

1083
AAUCACGCCAAGCUGCCGG
3839
1083
AAUCACGCCAAGCUGCCGG
3839
1101
CCGGCAGCUUGGCGUGAUU
3935

1101
GCCAGACCGCAAUGAGCCC
3840
1101
GCCAGACCGCAAUGAGCCC
3840
1119
GGGCUCAUUGCGGUCUGGC
3936

1119
CCACCGAAUCCAACAAAUC
3841
1119
CCACCGAAUCCAACAAAUC
3841
1137
GAUUUGUUGGAUUCGGUGG
3937

1137
CCUCAACUACAUCAAAGGG
3842
1137
CCUCAACUACAUCAAAGGG
3842
1155
CCCUUUGAUGUAGUUGAGG
3938

1155
GAAUCUGAAGCAUGUGGUC
3843
1155
GAAUCUGAAGCAUGUGGUC
3843
1173
GACCACAUGCUUCAGAUUC
3939

1173
CUAGUUGACAGAAAGAGAU
3844
1173
CUAGUUGACAGAAAGAGAU
3844
1191
AUCUCUUUCUGUCAACUAG
3940

1191
UCAGGUUUCCAGAGCUGAG
3845
1191
UCAGGUUUCCAGAGCUGAG
3845
1209
CUCAGCUCUGGAAACCUGA
3941

1209
GGAGUGGUGCCUAUAAUGA
3846
1209
GGAGUGGUGCCUAUAAUGA
3846
1227
UCAUUAUAGGCACCACUCC
3942

1227
AAGACAGCGUGUUUAUGCA
3847
1227
AAGACAGCGUGUUUAUGCA
3847
1245
UGCAUAAACACGCUGUCUU
3943

1245
AAGCAGUUUGUGGAAUUUG
3848
1245
AAGCAGUUUGUGGAAUUUG
3848
1263
CAAAUUCCACAAACUGCUU
3944

1263
GUGACUGCAGGGAAAAUUU
3849
1263
GUGACUGCAGGGAAAAUUU
3849
1281
AAAUUUUCCCUGCAGUCAC
3945

1281
UGAAAGAAAUUACUUCCUG
3850
1281
UGAAAGAAAUUACUUCCUG
3850
1299
CAGGAAGUAAUUUCUUUCA
3946

1299
GAAAAUUUCCAAGGGGCAU
3851
1299
GAAAAUUUCCAAGGGGCAU
3851
1317
AUGCCCCUUGGAAAUUUUC
3947

1317
UCAAGUGGCAGCUGGCUUC
3852
1317
UCAAGUGGCAGCUGGCUUC
3852
1335
GAAGCCAGCUGCCACUUGA
3948

1335
CCUGGGGUGAAGAGGCAGG
3853
1335
CCUGGGGUGAAGAGGCAGG
3853
1353
CCUGCCUCUUCACCCCAGG
3949

1353
GCACCCCAGAGUCCUCAAC
3854
1353
GCACCCCAGAGUCCUCAAC
3854
1371
GUUGAGGACUCUGGGGUGC
3950

1371
CUGGACCUAGGGGAAGAAG
3855
1371
CUGGACCUAGGGGAAGAAG
3855
1389
CUUCUUCCCCUAGGUCCAG
3951

1389
GGAGAUAUCCCACAUUUAA
3856
1389
GGAGAUAUCCCACAUUUAA
3856
1407
UUAAAUGUGGGAUAUCUCC
3952

1407
AAGUUCUUAUUUAAAAAAA
3857
1407
AAGUUCUUAUUUAAAAAAA
3857
1425
UUUUUUUAAAUAAGAACUU
3953

1425
ACACACACACACAAAUGAA
3858
1425
ACACACACACACAAAUGAA
3858
1443
UUCAUUUGUGUGUGUGUGU
3954

1443
AAUUUUUAAUCUUUGAAAA
3859
1443
AAUUUUUAAUCUUUGAAAA
3859
1461
UUUUCAAAGAUUAAAAAUU
3955

1461
AUUAUUUUUAAGCGAAUUG
3860
1461
AUUAUUUUUAAGCGAAUUG
3860
1479
CAAUUCGCUUAAAAAUAAU
3956

1479
GGGGAGGGGAGUAUUUUAA
3861
1479
GGGGAGGGGAGUAUUUUAA
3861
1497
UUAAAAUACUCCCCUCCCC
3957

1497
AUCAUCUUAAAUGAAACAG
3862
1497
AUCAUCUUAAAUGAAACAG
3862
1515
CUGUUUCAUUUAAGAUGAU
3958

1515
GAUCAGAAGCUGGAUGAGA
3863
1515
GAUCAGAAGCUGGAUGAGA
3863
1533
UCUCAUCCAGCUUCUGAUC
3959

1533
AGCAGUCACCAGUUUGUAG
3864
1533
AGCAGUCACCAGUUUGUAG
3864
1551
CUACAAACUGGUGACUGCU
3960

1551
GGGCAGGAGGCAGCUGAGA
3865
1551
GGGCAGGAGGCAGCUGAGA
3865
1569
UCUCAGCUGCCUCCUGCCC
3961

1569
AGGCAGGGUUUGGGCCUCA
3866
1569
AGGCAGGGUUUGGGCCUCA
3866
1587
UGAGGCCCAAACCCUGCCU
3962

1587
AGGACCAUCCAGGUGGAGC
3867
1587
AGGACCAUCCAGGUGGAGC
3867
1605
GCUCCACCUGGAUGGUCCU
3963

1605
CCCUGGGAGAGAGGGUACU
3868
1605
CCCUGGGAGAGAGGGUACU
3868
1623
AGUACCCUCUCUCCCAGGG
3964

1623
UGAUCAGCAGACUGGGAGG
3869
1623
UGAUCAGCAGACUGGGAGG
3869
1641
CCUCCCAGUCUGCUGAUCA
3965

1641
GUGGGGAGAAGUCCGCUGG
3870
1641
GUGGGGAGAAGUCCGCUGG
3870
1659
CCAGCGGACUUCUCCCCAC
3966

1659
GUGUUGUUUUAGUGUUAUA
3871
1659
GUGUUGUUUUAGUGUUAUA
3871
1677
UAUAACACUAAAACAACAC
3967

1677
AUAUCUUUGGUUUUUUUAA
3872
1677
AUAUCUUUGGUUUUUUUAA
3872
1695
UUAAAAAAACCAAAGAUAU
3968

1695
AUAAAAUCUUUGAAAACCU
3873
1695
AUAAAAUCUUUGAAAACCU
3873
1713
AGGUUUUCAAAGAUUUUAU
3969

1713
UAAAAAAAAAAAAAAAAAA
3874
1713
UAAAAAAAAAAAAAAAAAA
3874
1731
UUUUUUUUUUUUUUUUUUA
3970

HDAC9 transcript variant4: NM_178423.1

3
GGAAGAGAGGCACAGACAC
4083
3
GGAAGAGAGGCACAGACAC
4083
21
GUGUCUGUGCGUCUCUUCC
4341

21
CAGAUAGGAGAAGGGCACC
4084
21
CAGAUAGGAGAAGGGCACC
4084
39
GGUGCCCUUCUCCUAUCUG
4342

39
CGGCUGGAGCCACUUGCAG
4085
39
CGGCUGGAGCCACUUGCAG
4085
57
CUGCAAGUGGCUCCAGCCG
4343

57
GGACUGAGGGUUUUUGCAA
4086
57
GGACUGAGGGUUUUUGCAA
4086
75
UUGCAAAAACCCUCAGUCC
4344

75
ACAAAACCCUAGCAGCCUG
4087
75
ACAAAACCCUAGCAGCCUG
4087
93
CAGGCUGCUAGGGUUUUGU
4345

93
GAAGAACUCUAAGCCAGAU
4088
93
GAAGAACUCUAAGCCAGAU
4088
111
AUCUGGCUUAGAGUUCUUC
4346

111
UGGGGUGGCUGGACGAGAG
4089
111
UGGGGUGGCUGGACGAGAG
4089
129
CUCUCGUCCAGCCACCCCA
4347

129
GCAGCUCUUGGCUCAGCAA
4090
129
GCAGCUCUUGGCUCAGCAA
4090
147
UUGCUGAGCCAAGAGCUGC
4348

147
AAGAAUGCACAGUAUGAUC
4091
147
AAGAAUGCACAGUAUGAUC
4091
165
GAUCAUACUGUGCAUUCUU
4349

165
CAGCUCAGUGGAUGUGAAG
4092
165
CAGCUCAGUGGAUGUGAAG
4092
183
CUUCACAUCCACUGAGCUG
4350

183
GUCAGAAGUUCCUGUGGGC
4093
183
GUCAGAAGUUCCUGUGGGC
4093
201
GCCCACAGGAACUUCUGAC
4351

201
CCUGGAGCCCAUCUCACCU
4094
201
CCUGGAGCCCAUCUCACCU
4094
219
AGGUGAGAUGGGCUCCAGG
4352

219
UUUAGACCUAAGGACAGAC
4095
219
UUUAGACCUAAGGACAGAC
4095
237
GUCUGUCCUUAGGUCUAAA
4353

237
CCUCAGGAUGAUGAUGCCC
4096
237
CCUCAGGAUGAUGAUGCCC
4096
255
GGGCAUCAUCAUCCUGAGG
4354

255
CGUGGUGGACCCUGUUGUC
4097
255
CGUGGUGGACCCUGUUGUC
4097
273
GACAACAGGGUCCACCACG
4355

273
CCGUGAGAAGCAAUUGCAG
4098
273
CCGUGAGAAGCAAUUGGAG
4098
291
CUGCAAUUGCUUCUCACGG
4356

291
GCAGGAAUUACUUCUUAUC
4099
291
GCAGGAAUUACUUCUUAUC
4099
309
GAUAAGAAGUAAUUCCUGC
4357

309
CCAGCAGCAGCAACAAAUC
4100
309
CCAGCAGCAGCAACAAAUC
4100
327
GAUUUGUUGCUGCUGCUGG
4358

327
CCAGAAGCAGCUUCUGAUA
4101
327
CCAGAAGCAGCUUCUGAUA
4101
345
UAUCAGAAGCUGCUUCUGG
4359

345
AGCAGAGUUUCAGAAACAG
4102
345
AGCAGAGUUUCAGAAACAG
4102
363
CUGUUUCUGAAACUCUGCU
4360

363
GCAUGAGAACUUGACACGG
4103
363
GCAUGAGAACUUGACACGG
4103
381
CCGUGUCAAGUUCUCAUGC
4361

381
GCAGCACCAGGCUCAGCUU
4104
381
GCAGCACCAGGCUCAGCUU
4104
399
AAGCUGAGCCUGGUGCUGC
4362

399
UCAGGAGCAUAUCAAGGAA
4105
399
UCAGGAGCAUAUCAAGGAA
4105
417
UUCCUUGAUAUGCUCCUGA
4363

417
ACUUCUAGCCAUAAAACAG
4106
417
ACUUCUAGCCAUAAAACAG
4106
435
CUGUUUUAUGGCUAGAAGU
4364

435
GCAACAAGAACUCCUAGAA
4107
435
GCAACAAGAACUCCUAGAA
4107
453
UUCUAGGAGUUCUUGUUGC
4365

453
AAAGGAGCAGAAACUGGAG
4108
453
AAAGGAGCAGAAACUGGAG
4108
471
CUCCAGUUUCUGCUCCUUU
4366

471
GCAGCAGAGGCAAGAACAG
4109
471
GCAGCAGAGGCAAGAACAG
4109
489
CUGUUCUUGCCUCUGCUGC
4367

489
GGAAGUAGAGAGGCAUCGC
4110
489
GGAAGUAGAGAGGCAUCGC
4110
507
GCGAUGCCUCUCUACUUCC
4368

507
CAGAGAACAGCAGCUUCCU
4111
507
CAGAGAACAGCAGCUUCCU
4111
525
AGGAAGCUGCUGUUCUCUG
4369

525
UCCUCUCAGAGGCAAAGAU
4112
525
UCCUCUCAGAGGGAAAGAU
4112
543
AUCUUUGCCUCUGAGAGGA
4370

543
UAGAGGACGAGAAAGGGCA
4113
543
UAGAGGACGAGAAAGGGCA
4113
561
UGCCCUUUCUCGUCCUCUA
4371

561
AGUGGCAAGUACAGAAGUA
4114
561
AGUGGCAAGUACAGAAGUA
4114
579
UACUUCUGUACUUGCCACU
4372

579
AAAGCAGAAGCUUCAAGAG
4115
579
AAAGCAGAAGCUUCAAGAG
4115
597
CUCUUGAAGCUUCUGCUUU
4373

597
GUUCCUACUGAGUAAAUCA
4116
597
GUUCCUACUGAGUAAAUCA
4116
615
UGAUUUACUCAGUAGGAAC
4374

615
AGCAACGAAAGACACUCCA
4117
615
AGCAACGAAAGACACUCCA
4117
633
UGGAGUGUCUUUCGUUGCU
4375

633
AACUAAUGGAAAAAAUCAU
4118
633
AACUAAUGGAAAAAAUCAU
4118
651
AUGAUUUUUUCCAUUAGUU
4376

651
UUCCGUGAGCCGCCAUCCC
4119
651
UUCCGUGAGCCGCCAUCCC
4119
669
GGGAUGGCGGCUCACGGAA
4377

669
CAAGCUCUGGUACACGGCU
4120
669
CAAGCUCUGGUACACGGCU
4120
687
AGCCGUGUACCAGAGCUUG
4378

687
UGCCCACCACACAUCAUUG
4121
687
UGCCCACCACACAUCAUUG
4121
705
CAAUGAUGUGUGGUGGGCA
4379

705
GGAUCAAAGCUCUCCACCC
4122
705
GGAUCAAAGCUCUCCACCC
4122
723
GGGUGGAGAGCUUUGAUCC
4380

723
CCUUAGUGGAACAUCUCCA
4123
723
CCUUAGUGGAACAUCUCCA
4123
741
UGGAGAUGUUCCACUAAGG
4381

741
AUCCUACAAGUACACAUUA
4124
741
AUCCUACAAGUACACAUUA
4124
759
UAAUGUGUACUUGUAGGAU
4382

759
ACCAGGAGCACAAGAUGCA
4125
759
ACCAGGAGCACAAGAUGCA
4125
777
UGCAUCUUGUGCUCCUGGU
4383

777
AAAGGAUGAUUUCCCCCUU
4126
777
AAAGGAUGAUUUCCCCCUU
4126
795
AAGGGGGAAAUCAUCCUUU
4384

795
UCGAAAAACUGCCUCUGAG
4127
795
UCGAAAAACUGCCUCUGAG
4127
813
CUCAGAGGCAGUUUUUCGA
4385

813
GCCCAACUUGAAGGUGCGG
4128
813
GCCCAACUUGAAGGUGCGG
4128
831
CCGCACCUUCAAGUUGGGC
4386

831
GUCCAGGUUAAAACAGAAA
4129
831
GUCCAGGUUAAAACAGAAA
4129
849
UUUCUGUUUUAACCUGGAC
4387

849
AGUGGCAGAGAGGAGAAGC
4130
849
AGUGGCAGAGAGGAGAAGC
4130
867
GCUUCUCCUCUCUGCCACU
4388

867
CAGCCCCUUACUCAGGCGG
4131
867
CAGCCCCUUACUCAGGCGG
4131
885
CCGCCUGAGUAAGGGGCUG
4389

885
GAAGGAUGGAAAUGUUGUC
4132
885
GAAGGAUGGAAAUGUUGUC
4132
903
GACAACAUUUCCAUCCUUC
4390

903
CACUUCAUUCAAGAAGCGA
4133
903
CACUUCAUUCAAGAAGCGA
4133
921
UCGCUUCUUGAAUGAAGUG
4391

921
AAUGUUUGAGGUGACAGAA
4134
921
AAUGUUUGAGGUGACAGAA
4134
939
UUCUGUCACCUCAAACAUU
4392

939
AUCCUCAGUCAGUAGCAGU
4135
939
AUCCUCAGUCAGUAGCAGU
4135
957
ACUGCUACUGACUGAGGAU
4393

957
UUCUCCAGGCUCUGGUCCC
4136
957
UUCUCCAGGCUCUGGUCCC
4136
975
GGGACCAGAGCCUGGAGAA
4394

975
CAGUUCACCAAACAAUGGG
4137
975
CAGUUCACCAAACAAUGGG
4137
993
CCCAUUGUUUGGUGAACUG
4395

993
GCCAACUGGAAGUGUUACU
4138
993
GCCAACUGGAAGUGUUACU
4138
1011
AGUAACACUUCCAGUUGGC
4396

1011
UGAAAAUGAGACUUCGGUU
4139
1011
UGAAAAUGAGACUUCGGUU
4139
1029
AACCGAAGUCUCAUUUUCA
4397

1029
UUUGCCCCCUACCCCUCAU
4140
1029
UUUGCCCCCUACCCCUCAU
4140
1047
AUGAGGGGUAGGGGGCAAA
4398

1047
UGCCGAGCAAAUGGUUUCA
4141
1047
UGCCGAGCAAAUGGUUUCA
4141
1065
UGAAACCAUUUGCUCGGCA
4399

1065
ACAGCAACGCAUUCUAAUU
4142
1065
ACAGCAACGCAUUCUAAUU
4142
1083
AAUUAGAAUGCGUUGCUGU
4400

1083
UCAUGAAGAUUCCAUGAAC
4143
1083
UCAUGAAGAUUCCAUGAAC
4143
1101
GUUCAUGGAAUCUUCAUGA
4401

1101
CCUGCUAAGUCUUUAUACC
4144
1101
CCUGCUAAGUCUUUAUACC
4144
1119
GGUAUAAAGACUUAGCAGG
4402

1119
CUCUCCUUCUUUGCCCAAC
4145
1119
CUCUCCUUCUUUGCCCAAC
4145
1137
GUUGGGCAAAGAAGGAGAG
4403

1137
CAUUACCUUGGGGCUUCCC
4146
1137
CAUUACCUUGGGGCUUCCC
4146
1155
GGGAAGCCCCAAGGUAAUG
4404

1155
CGCAGUGCCAUCCCAGCUC
4147
1155
CGCAGUGCCAUCCCAGCUC
4147
1173
GAGCUGGGAUGGCACUGCG
4405

1173
CAAUGCUUCGAAUUCACUC
4148
1173
CAAUGCUUCGAAUUCACUC
4148
1191
GAGUGAAUUCGAAGCAUUG
4406

1191
CAAAGAAAAGCAGAAGUGU
4149
1191
CAAAGAAAAGCAGAAGUGU
4149
1209
ACACUUCUGCUUUUCUUUG
4407

1209
UGAGACGCAGACGCUUAGG
4150
1209
UGAGACGCAGACGCUUAGG
4150
1227
CCUAAGCGUCUGCGUCUCA
4408

1227
GCAAGGUGUUCCUCUGCCU
4151
1227
GCAAGGUGUUCCUCUGCCU
4151
1245
AGGCAGAGGAACACCUUGC
4409

1245
UGGGCAGUAUGGAGGCAGC
4152
1245
UGGGCAGUAUGGAGGCAGC
4152
1263
GCUGCCUCCAUACUGCCCA
4410

1263
CAUCCCGGCAUCUUCCAGC
4153
1263
CAUCCCGGCAUCUUCCAGC
4153
1281
GCUGGAAGAUGCCGGGAUG
4411

1281
CCACCCUCAUGUUACUUUA
4154
1281
CCACCCUCAUGUUACUUUA
4154
1299
UAAAGUAACAUGAGGGUGG
4412

1299
AGAGGGAAAGCCACCCAAC
4155
1299
AGAGGGAAAGCCACCCAAC
4155
1317
GUUGGGUGGCUUUCCCUCU
4413

1317
CAGCAGGCACCAGGGUCUC
4156
1317
CAGCAGCCACCAGGCUCUC
4156
1335
GAGAGCCUGGUGGCUGCUG
4414

1335
CCUGCAGCAUUUAUUAUUG
4157
1335
CCUGCAGCAUUUAUUAUUG
4157
1353
CAAUAAUAAAUGCUGCAGG
4415

1353
GAAAGAACAAAUGCGACAG
4158
1353
GAAAGAACAAAUGCGACAG
4158
1371
CUGUCGCAUUUGUUCUUUC
4416

1371
GCAAAAGCUUCUUGUAGCU
4159
1371
GCAAAAGCUUCUUGUAGCU
4159
1389
AGCUACAAGAAGCUUUUGC
4417

1389
UGGUGGAGUUCCCUUACAU
4160
1389
UGGUGGAGUUCCCUUACAU
4160
1407
AUGUAAGGGAACUCCACCA
4418

1407
UCCUCAGUCUCCCUUGGCA
4161
1407
UCCUCAGUCUCCCUUGGCA
4161
1425
UGCCAAGGGAGACUGAGGA
4419

1425
AACAAAAGAGAGAAUUUCA
4162
1425
AACAAAAGAGAGAAUUUCA
4162
1443
UGAAAUUCUCUCUUUUGUU
4420

1443
ACCUGGCAUUAGAGGUACC
4163
1443
ACCUGGCAUUAGAGGUACC
4163
1461
GGUACCUCUAAUGCCAGGU
4421

1461
CCACAAAUUGCCCCGUCAC
4164
1461
CCACAAAUUGCCCCGUCAC
4164
1479
GUGACGGGGCAAUUUGUGG
4422

1479
CAGACCCCUGAACCGAACC
4165
1479
CAGACCCCUGAACCGAACC
4165
1497
GGUUCGGUUCAGGGGUCUG
4423

1497
CCAGUCUGCACCUUUGCCU
4166
1497
CCAGUCUGCACCUUUGCCU
4166
1515
AGGCAAAGGUGCAGACUGG
4424

1515
UCAGAGCACGUUGGCUCAG
4167
1515
UCAGAGCACGUUGGCUCAG
4167
1533
CUGAGCCAACGUGCUCUGA
4425

1533
GCUGGUCAUUCAACAGCAA
4168
1533
GCUGGUCAUUCAACAGCAA
4168
1551
UUGCUGUUGAAUGACCAGC
4426

1551
ACACCAGCAAUUCUUGGAG
4169
1551
ACACCAGCAAUUCUUGGAG
4169
1569
CUCCAAGAAUUGCUGGUGU
4427

1569
GAAGCAGAAGCAAUACCAG
4170
1569
GAAGCAGAAGCAAUACCAG
4170
1587
CUGGUAUUGCUUCUGCUUC
4428

1587
GCAGCAGAUCCACAUGAAC
4171
1587
GCAGCAGAUCCACAUGAAC
4171
1605
GUUCAUGUGGAUCUGCUGC
4429

1605
CAAACUGCUUUCGAAAUCU
4172
1605
CAAACUGCUUUCGAAAUCU
4172
1623
AGAUUUCGAAAGCAGUUUG
4430

1623
UAUUGAACAACUGAAGCAA
4173
1623
UAUUGAACAACUGAAGCAA
4173
1641
UUGCUUCAGUUGUUCAAUA
4431

1641
ACCAGGCAGUCACCUUGAG
4174
1641
ACCAGGCAGUCACCUUGAG
4174
1659
CUCAAGGUGACUGCCUGGU
4432

1659
GGAAGCAGAGGAAGAGCUU
4175
1659
GGAAGCAGAGGAAGAGCUU
4175
1677
AAGCUCUUCCUCUGCUUCC
4433

1677
UCAGGGGGACCAGGCGAUG
4176
1677
UCAGGGGGACCAGGCGAUG
4176
1695
CAUCGCCUGGUCCCCCUGA
4434

1695
GCAGGAAGACAGAGCGCCC
4177
1695
GCAGGAAGACAGAGCGCCC
4177
1713
GGGCGCUCUGUCUUCCUGC
4435

1713
CUCUAGUGGCAACAGCACU
4178
1713
CUCUAGUGGCAACAGCACU
4178
1731
AGUGCUGUUGCCACUAGAG
4436

1731
UAGGAGCGACAGCAGUGCU
4179
1731
UAGGAGCGACAGCAGUGCU
4179
1749
AGCACUGCUGUCGCUCCUA
4437

1749
UUGUGUGGAUGACACACUG
4180
1749
UUGUGUGGAUGACACACUG
4180
1767
CAGUGUGUCAUCCACACAA
4438

1767
GGGACAAGUUGGGGCUGUG
4181
1767
GGGACAAGUUGGGGCUGUG
4181
1785
CACAGCCCCAACUUGUCCC
4439

1785
GAAGGUCAAGGAGGAACCA
4182
1785
GAAGGUCAAGGAGGAACCA
4182
1803
UGGUUCCUCCUUGACCUUC
4440

1803
AGUGGACAGUGAUGAAGAU
4183
1803
AGUGGACAGUGAUGAAGAU
4183
1821
AUCUUCAUCACUGUCCACU
4441

1821
UGCUCAGAUCCAGGAAAUG
4184
1821
UGCUCAGAUCCAGGAAAUG
4184
1839
CAUUUCCUGGAUCUGAGCA
4442

1839
GGAAUCUGGGGAGCAGGCU
4185
1839
GGAAUCUGGGGAGCAGGCU
4185
1857
AGCCUGCUCCCCAGAUUCC
4443

1857
UGCUUUUAUGCAACAGCCU
4186
1857
UGCUUUUAUGCAACAGCCU
4186
1875
AGGCUGUUGCAUAAAAGCA
4444

1875
UUUCCUGGAACCCACGCAC
4187
1875
UUUCCUGGAACCCACGCAC
4187
1893
GUGCGUGGGUUCCAGGAAA
4445

1893
CACACGUGCGCUCUCUGUG
4188
1893
CACACGUGCGCUCUCUGUG
4188
1911
CACAGAGAGCGCACGUGUG
4446

1911
GCGCCAAGCUCCGCUGGCU
4189
1911
GCGCCAAGCUCCGCUGGCU
4189
1929
AGCCAGCGGAGCUUGGCGC
4447

1929
UGCGGUUGGCAUGGAUGGA
4190
1929
UGCGGUUGGCAUGGAUGGA
4190
1947
UCCAUCCAUGCCAACCGCA
4448

1947
AUUAGAGAAACACCGUCUC
4191
1947
AUUAGAGAAACACCGUCUC
4191
1965
GAGACGGUGUUUCUCUAAU
4449

1965
CGUCUCCAGGACUCACUCU
4192
1965
GGUCUCCAGGACUCACUCU
4192
1983
AGAGUGAGUCCUGGAGACG
4450

1983
UUCCCCUGCUGCCUCUGUU
4193
1983
UUCCCCUGCUGCCUCUGUU
4193
2001
AACAGAGGCAGGAGGGGAA
4451

2001
UUUACCUCACCCAGCAAUG
4194
2001
UUUACCUCACCCAGCAAUG
4194
2019
CAUUGCUGGGUGAGGUAAA
4452

2019
GGACCGCCCCCUCCAGCCU
4195
2019
GGACCGCCCCCUCCAGCCU
4195
2037
AGGCUGGAGGGGGCGGUCC
4453

2037
UGGCUCUGCAACUGGAAUU
4196
2037
UGGCUCUGCAACUGGAAUU
4196
2055
AAUUCCAGUUGCAGAGCCA
4454

2055
UGCCUAUGACCCCUUGAUG
4197
2055
UGCCUAUGACCCCUUGAUG
4197
2073
CAUCAAGGGGUCAUAGGCA
4455

2073
GCUGAAACACCAGUGCGUU
4198
2073
GCUGAAACACCAGUGCGUU
4198
2091
AACGCACUGGUGUUUCAGC
4456

2091
UUGUGGCAAUUCCACCACC
4199
2091
UUGUGGCAAUUCCACCACC
4199
2109
GGUGGUGGAAUUGCCACAA
4457

2109
CCACCCUGAGCAUGCUGGA
4200
2109
CCACCCUGAGCAUGCUGGA
4200
2127
UCCAGCAUGCUCAGGGUGG
4458

2127
ACGAAUACAGAGUAUCUGG
4201
2127
ACGAAUACAGAGUAUCUGG
4201
2145
CCAGAUACUCUGUAUUCGU
4459

2145
GUCACGACUGCAAGAAACU
4202
2145
GUCACGACUGCAAGAAACU
4202
2163
AGUUUCUUGCAGUCGUGAC
4460

2163
UGGGCUGCUAAAUAAAUGU
4203
2163
UGGGCUGCUAAAUAAAUGU
4203
2181
ACAUUUAUUUAGCAGCCCA
4461

2181
UGAGCGAAUUCAAGGUCGA
4204
2181
UGAGCGAAUUCAAGGUCGA
4204
2199
UCGACCUUGAAUUCGCUCA
4462

2199
AAAAGCCAGCCUGGAGGAA
4205
2199
AAAAGCCAGCCUGGAGGAA
4205
2217
UUCCUCCAGGCUGGCUUUU
4463

2217
AAUACAGCUUGUUCAUUCU
4206
2217
AAUACAGCUUGUUCAUUCU
4206
2235
AGAAUGAACAAGCUGUAUU
4464

2235
UGAACAUCACUCACUGUUG
4207
2235
UGAACAUCACUCACUGUUG
4207
2253
CAACAGUGAGUGAUGUUCA
4465

2253
GUAUGGCACCAACCCCCUG
4208
2253
GUAUGGCACCAACCCCCUG
4208
2271
CAGGGGGUUGGUGCCAUAC
4466

2271
GGACGGACAGAAGCUGGAC
4209
2271
GGACGGACAGAAGCUGGAC
4209
2289
GUCCAGCUUCUGUCCGUCC
4467

2289
CCCCAGGAUACUCCUAGGU
4210
2289
CCCCAGGAUACUCCUAGGU
4210
2307
ACCUAGGAGUAUCCUGGGG
4468

2307
UGAUGACUCUCAAAAGUUU
4211
2307
UGAUGACUCUCAAAAGUUU
4211
2325
AAACUUUUGAGAGUCAUCA
4469

2325
UUUUUCCUCAUUACCUUGU
4212
2325
UUUUUCCUCAUUACCUUGU
4212
2343
ACAAGGUAAUGAGGAAAAA
4470

2343
UGGUGGACUUGGGGUGGAC
4213
2343
UGGUGGACUUGGGGUGGAC
4213
2361
GUCCACCCCAAGUCCACCA
4471

2361
CAGUGACACCAUUUGGAAU
4214
2361
CAGUGACACCAUUUGGAAU
4214
2379
AUUCCAAAUGGUGUCACUG
4472

2379
UGAGCUACACUCGUCCGGU
4215
2379
UGAGCUACACUCGUCCGGU
4215
2397
ACCGGAGGAGUGUAGGUCA
4473

2397
UGCUGCACGCAUGGCUGUU
4216
2397
UGCUGCACGCAUGGCUGUU
4216
2415
AACAGCCAUGCGUGCAGCA
4474

2415
UGGCUGUGUCAUCGAGCUG
4217
2415
UGGCUGUGUCAUCGAGCUG
4217
2433
CAGCUCGAUGACACAGCCA
4475

2433
GGCUUCCAAAGUGGCCUCA
4218
2433
GGCUUCCAAAGUGGCCUCA
4218
2451
UGAGGCCACUUUGGAAGCC
4476

2451
AGGAGAGCUGAAGAAUGGG
4219
2451
AGGAGAGCUGAAGAAUGGG
4219
2469
CCCAUUCUUCAGCUCUCCU
4477

2469
GUUUGCUGUUGUGAGGCCC
4220
2469
GUUUGCUGUUGUGAGGCCC
4220
2487
GGGCCUCACAACAGCAAAC
4478

2487
CCCUGGCCAUCACGCUGAA
4221
2487
CCCUGGCCAUCACGCUGAA
4221
2505
UUCAGCGUGAUGGCCAGGG
4479

2505
AGAAUCCACAGCCAUGGGG
4222
2505
AGAAUCCACAGCCAUGGGG
4222
2523
CCCCAUGGCUGUGGAUUCU
4480

2523
GUUCUGCUUUUUUAAUUCA
4223
2523
GUUCUGCUUUUUUAAUUCA
4223
2541
UGAAUUAAAAAAGCAGAAC
4481

2541
AGUUGCAAUUACCGCCAAA
4224
2541
AGUUGCAAUUACCGCCAAA
4224
2559
UUUGGCGGUAAUUGCAACU
4482

2559
AUACUUGAGAGACCAACUA
4225
2559
AUACUUGAGAGACCAACUA
4225
2577
UAGUUGGUCUCUCAAGUAU
4483

2577
AAAUAUAAGCAAGAUAUUG
4226
2577
AAAUAUAAGCAAGAUAUUG
4226
2595
CAAUAUCUUGCUUAUAUUU
4484

2595
GAUUGUAGAUCUGGAUGUU
4227
2595
GAUUGUAGAUCUGGAUGUU
4227
2613
AACAUCCAGAUCUACAAUC
4485

2613
UCACCAUGGAAACGGUACC
4228
2613
UCACCAUGGAAACGGUACC
4228
2631
GGUACCGUUUCCAUGGUGA
4486

2631
CCAGCAGGCCUUUUAUGCU
4229
2631
CCAGCAGGCCUUUUAUGCU
4229
2649
AGCAUAAAAGGCCUGCUGG
4487

2649
UGACCCCAGGAUCCUGUAC
4230
2649
UGACCCCAGCAUCCUGUAC
4230
2667
GUACAGGAUGCUGGGGUCA
4488

2667
CAUUUCACUCCAUCGCUAU
4231
2667
CAUUUCACUCCAUCGCUAU
4231
2685
AUAGCGAUGGAGUGAAAUG
4489

2685
UGAUGAAGGGAACUUUUUC
4232
2685
UGAUGAAGGGAACUUUUUC
4232
2703
GAAAAAGUUCCCUUCAUCA
4490

2703
CCCUGGCAGUGGAGCCCCA
4233
2703
CCCUGGCAGUGGAGCCCCA
4233
2721
UGGGGCUCCACUGCCAGGG
4491

2721
AAAUGAGGUUGGAACAGGC
4234
2721
AAAUGAGGUUGGAACAGGC
4234
2739
GCCUGUUCCAACCUCAUUU
4492

2739
CCUUGGAGAAGGGUACAAU
4235
2739
CCUUGGAGAAGGGUACAAU
4235
2757
AUUGUACCCUUCUCCAAGG
4493

2757
UAUAAAUAUUGCCUGGACA
4236
2757
UAUAAAUAUUGCCUGGACA
4236
2775
UGUCCAGGCAAUAUUUAUA
4494

2775
AGGUGGCCUUGAUCCUCCC
4237
2775
AGGUGGCCUUGAUCCUCCC
4237
2793
GGGAGGAUCAAGGCCACCU
4495

2793
CAUGGGAGAUGUUGAGUAC
4238
2793
CAUGGGAGAUGUUGAGUAC
4238
2811
GUACUCAACAUCUCCCAUG
4496

2811
CCUUGAAGCAUUCAGGACC
4239
2811
CCUUGAAGCAUUCAGGACC
4239
2829
GGUCCUGAAUGCUUCAAGG
4497

2829
CAUCGUGAAGCCUGUGGCC
4240
2829
CAUCGUGAAGCCUGUGGCC
4240
2847
GGCCACAGGCUUCACGAUG
4498

2847
CAAAGAGUUUGAUCCAGAC
4241
2847
CAAAGAGUUUGAUCCAGAC
4241
2865
GUCUGGAUCAAACUCUUUG
4499

2865
CAUGGUCUUAGUAUCUGCU
4242
2865
CAUGGUCUUAGUAUCUGCU
4242
2883
AGCAGAUACUAAGACCAUG
4500

2883
UGGAUUUGAUGCAUUGGAA
4243
2883
UGGAUUUGAUGCAUUGGAA
4243
2901
UUCCAAUGCAUCAAAUCCA
4501

2901
AGGCCACACCCCUCCUCUA
4244
2901
AGGCCACACCCCUCCUCUA
4244
2919
UAGAGGAGGGGUGUGGCCU
4502

2919
AGGAGGGUACAAAGUGACG
4245
2919
AGGAGGGUACAAAGUGACG
4245
2937
CGUCACUUUGUACCCUCCU
4503

2937
GGCAAAAUGUUUUGGUCAU
4246
2937
GGCAAAAUGUUUUGGUCAU
4246
2955
AUGACCAAAACAUUUUGCC
4504

2955
UUUGACGAAGCAAUUGAUG
4247
2955
UUUGACGAAGCAAUUGAUG
4247
2973
CAUCAAUUGCUUCGUCAAA
4505

2973
GACAUUGGCUGAUGGACGU
4248
2973
GACAUUGGCUGAUGGACGU
4248
2991
ACGUCCAUCAGCCAAUGUC
4506

2991
UGUGGUGUUGGCUCUAGAA
4249
2991
UGUGGUGUUGGCUCUAGAA
4249
3009
UUCUAGAGCCAACACCACA
4507

3009
AGGAGGACAUGAUCUCACA
4250
3009
AGGAGGACAUGAUCUCACA
4250
3027
UGUGAGAUCAUGUCCUCCU
4508

3027
AGCCAUCUGUGAUGCAUCA
4251
3027
AGCCAUCUGUGAUGCAUCA
4251
3045
UGAUGCAUCACAGAUGGCU
4509

3045
AGAAGCCUGUGUAAAUGCC
4252
3045
AGAAGCCUGUGUAAAUGCC
4252
3063
GGCAUUUACACAGGCUUCU
4510

3063
CCUUCUAGGAAAUGAGCUG
4253
3063
CCUUCUAGGAAAUGAGCUG
4253
3081
CAGCUCAUUUCCUAGAAGG
4511

3081
GGAGCCACUUGCAGAAGAU
4254
3081
GGAGCCACUUGCAGAAGAU
4254
3099
AUCUUCUGCAAGUGGCUCC
4512

3099
UAUUCUCCACCAAAGCCCG
4255
3099
UAUUCUCCACCAAAGCCCG
4255
3117
CGGGCUUUGGUGGAGAAUA
4513

3117
GAAUAUGAAUGCUGUUAUU
4256
3117
GAAUAUGAAUGCUGUUAUU
4256
3135
AAUAACAGCAUUCAUAUUC
4514

3135
UUCUUUACAGAAGAUCAUU
4257
3135
UUCUUUACAGAAGAUCAUU
4257
3153
AAUGAUCUUCUGUAAAGAA
4515

3153
UGAAAUUCAAAGCAAGUAU
4258
3153
UGAAAUUCAAAGCAAGUAU
4258
3171
AUACUUGCUUUGAAUUUCA
4516

3171
UUGGAAGUCAGUAAGGAUG
4259
3171
UUGGAAGUCAGUAAGGAUG
4259
3189
CAUCCUUACUGACUUCCAA
4517

3189
GGUGGCUGUGCCAAGGGGC
4260
3189
GGUGGCUGUGCCAAGGGGC
4260
3207
GCCCCUUGGCACAGCCACC
4518

3207
CUGUGCUCUGGCUGGUGCU
4261
3207
CUGUGCUCUGGCUGGUGCU
4261
3225
AGCACCAGCCAGAGCACAG
4519

3225
UCAGUUGCAAGAGGAGACA
4262
3225
UCAGUUGCAAGAGGAGACA
4262
3243
UGUCUCCUCUUGCAACUGA
4520

3243
AGAGACCGUUUCUGCCCUG
4263
3243
AGAGACCGUUUCUGCCCUG
4263
3261
CAGGGCAGkAACGGUCUCU
4521

3261
GGCCUCCCUAACAGUGGAU
4264
3261
GGCCUCCCUAACAGUGGAU
4264
3279
AUCCACUGUUAGGGAGGCC
4522

3279
UGUGGAACAGCCCUUUGCU
4265
3279
UGUGGAACAGCCCUUUGCU
4265
3297
AGCAAAGGGCUGUUCCACA
4523

3297
UCAGGAAGACAGCAGAACU
4266
3297
UCAGGAAGACAGCAGAACU
4266
3315
AGUUCUGCUGUCUUCCUGA
4524

3315
UGCUGGUGAGCCUAUGGAA
4267
3315
UGCUGGUGAGCCUAUGGAA
4267
3333
UUCCAUAGGCUCACCAGCA
4525

3333
AGAGGAGCCAGCCUUGUGA
4268
3333
AGAGGAGCCAGCCUUGUGA
4268
3351
UCACAAGGCUGGCUCCUCU
4526

3351
AAGUGCCAAGUCCCCCUCU
4269
3351
AAGUGCCAAGUCCCCCUCU
4269
3369
AGAGGGGGACUUGGCACUU
4527

3369
UGAUAUUUCCUGUGUGUGA
4270
3369
UGAUAUUUCCUGUGUGUGA
4270
3387
UCACACACAGGAAAUAUCA
4528

3387
ACAUCAUUGUGUAUCCCCC
4271
3387
ACAUCAUUGUGUAUCCCCC
4271
3405
GGGGGAUACACAAUGAUGU
4529

3405
CCACCCCAGUACCCUCAGA
4272
3405
CCACCCCAGUACCCUCAGA
4272
3423
UCUGAGGGUACUGGGGUGG
4530

3423
ACAUGUCUUGUCUGCUGCC
4273
3423
ACAUGUCUUGUCUGCUGCC
4273
3441
GGCAGCAGACAAGACAUGU
4531

3441
CUGGGUGGCACAGAUUCAA
4274
3441
CUGGGUGGCACAGAUUCAA
4274
3459
UUGAAUCUGUGCCACCCAG
4532

3459
AUGGAACAUAAACACUGGG
4275
3459
AUGGAACAUAAACACUGGG
4275
3477
CCCAGUGUUUAUGUUCCAU
4533

3477
GCACAAAAUUCUGAACAGC
4276
3477
GCACAAAAUUCUGAACAGC
4276
3495
GCUGUUCAGAAUUUUGUGC
4534

3495
CAGCUUCACUUGUUCUUUG
4277
3495
CAGCUUCACUUGUUCUUUG
4277
3513
CAAAGAACAAGUGAAGCUG
4535

3513
GGAUGGACUUGAAAGGGCA
4278
3513
GGAUGGACUUGAAAGGGCA
4278
3531
UGCCCUUUCAAGUCCAUCC
4536

3531
AUUAAAGAUUCCUUAAACG
4279
3531
AUUAAAGAUUCCUUAAACG
4279
3549
CGUUUAAGGAAUCUUUAAU
4537

3549
GUAACCGCUGUGAUUCUAG
4280
3549
GUAACCGCUGUGAUUCUAG
4280
3567
CUAGAAUCACAGCGGUUAC
4538

3567
GAGUUACAGUAAACCACGA
4281
3567
GAGUUACAGUAAACCACGA
4281
3585
UCGUGGUUUACUGUAACUC
4539

3585
AUUGGAAGAAACUGCUUCC
4282
3585
AUUGGAAGAAACUGCUUCC
4282
3603
GGAAGCAGUUUCUUCCAAU
4540

3603
CAGCAUGCUUUUAAUAUGC
4283
3603
CAGCAUGCUUUUAAUAUGC
4283
3621
GCAUAUUAAAAGCAUGCUG
4541

3621
CUGGGUGACCCACUCCUAG
4284
3621
CUGGGUGACCCACUCCUAG
4284
3639
CUAGGAGUGGGUCACCCAG
4542

3639
GACACCAAGUUUGAACUAG
4285
3639
GACACCAAGUUUGAACUAG
4285
3657
CUAGUUCAAACUUGGUGUC
4543

3657
GAAACAUUCAGUACAGCAC
4286
3657
GAAACAUUCAGUACAGCAC
4286
3675
GUGCUGUACUGAAUGUUUC
4544

3675
CUAGAUAUUGUUAAUUUCA
4287
3675
CUAGAUAUUGUUAAUUUCA
4287
3693
UGAAAUUAACAAUAUCUAG
4545

3693
AGAAGCUAUGACAGCCAGU
4288
3693
AGAAGCUAUGACAGCCAGU
4288
3711
ACUGGCUGUCAUAGCUUCU
4546

3711
UGAAAUUUUGGGCAAAACC
4289
3711
UGAAAUUUUGGGCAAAACC
4289
3729
GGUUUUGCCCAAAAUUUCA
4547

3729
CUGAGACAUAGUCAUUCCU
4290
3729
CUGAGACAUAGUCAUUCCU
4290
3747
AGGAAUGACUAUGUCUCAG
4548

3747
UGACAUUCUGAUCAGCUUU
4291
3747
UGACAUUCUGAUCAGCUUU
4291
3765
AAAGCUGAUCAGAAUGUCA
4549

3765
UUUUUGGGGUAAUUUGUUU
4292
3765
UUUUUGGGGUAAUUUGUUU
4292
3783
AAACAAAUUACCCCAAAAA
4550

3783
UUUCAAACAGUCUUAACUU
4293
3783
UUUCAAACAGUCUUAACUU
4293
3801
AAGUUAAGACUGUUUGAAA
4551

3801
UGUUUACAAGAUUUGCUUU
4294
3801
UGUUUACAAGAUUUGCUUU
4294
3819
AAAGCAAAUCUUGUAAACA
4552

3819
UUAGCUAUGAACGGAUCGU
4295
3819
UUAGCUAUGAACGGAUCGU
4295
3837
ACGAUCCGUUCAUAGCUAA
4553

3837
UAAUUCCACCCAGAAUGUA
4296
3837
UAAUUCCACCCAGAAUGUA
4296
3855
UACAUUCUGGGUGGAAUUA
4554

3855
AAUGUUUCUUGUUUGUUUG
4297
3855
AAUGUUUCUUGUUUGUUUG
4297
3873
CAAACAAACAAGAAACAUU
4555

3873
GUUUUGUUUUGUUAGGGUU
4298
3873
GUUUUGUUUUGUUAGGGUU
4298
3891
AACCCUAACPAAACAAAAC
4556

3891
UUUUUUCUCAACUUUAACA
4299
3891
UUUUUUCUCAACUUUAACA
4299
3909
UGUUAAAGUUGAGAAAAAA
4557

3909
ACACAGUUCAACUGUUCCU
4300
3909
ACACAGUUCAACUGUUCCU
4300
3927
AGGAACAGUUGAACUGUGU
4558

3927
UAGUAAAAGUUCAAGAUGG
4301
3927
UAGUAAAAGUUCAAGAUGG
4301
3945
CCAUCUUGAACUUUUACUA
4559

3945
GAGGAACUAGCAUGAGGCU
4302
3945
GAGGAACUAGCAUGAGGCU
4302
3963
AGCCUCAUGGUAGUUCCUC
4560

3963
UUUUUUCAGUAUCUCGAAG
4303
3963
UUUUUUCAGUAUCUCGAAG
4303
3981
CUUCGAGAUACUGAAAAAA
4561

3981
GUCCAAAUGCCAAAGGAAC
4304
3981
GUCCAAAUGCCAAAGGAAC
4304
3999
GUUCCUUUGGCAUUUGGAC
4562

3999
CCUCACACACUGUUUGUAA
4305
3999
CCUCACACACUGUUUGUAA
4305
4017
UUACAAACAGUGUGUGAGG
4563

4017
AUGGUGCAAUAUUUUAUAU
4306
4017
AUGGUGCAAUAUUUUAUAU
4306
4035
AUAUAAAAUAUUGCACCAU
4564

4035
UCACUUUUUUUUAAACAUC
4307
4035
UCACUUUUUUUUAAACAUC
4307
4053
GAUGUUUAAAAAAAAGUGA
4565

4053
CCCCAACAUCUUUGUGUUC
4308
4053
CCCCAACAUCUUUGUGUUC
4308
4071
GAACACAAAGAUGUUGGGG
4566

4071
CUCACACACAGGCAAUUUG
4309
4071
CUCACACACAGGCAAUUUG
4309
4089
CAAAUUGCCUGUGUGUGAG
4567

4089
GCAAUGUUGCAAUUGUGUU
4310
4089
GCAAUGUUGCAAUUGUGUU
4310
4107
AACACAAUUGCAACAUUGC
4568

4107
UGGAGAAUGAAGUCCCCCC
4311
4107
UGGAGAAUGAAGUCCCCCC
4311
4125
GGGGGGACUUCAUUCUCCA
4569

4125
CACCUCCCAGCCACACACA
4312
4125
CACCUCCCAGCCACACACA
4312
4143
UGUGUGUGGCUGGGAGGUG
4570

4143
ACAUCCUUUGUUCUCAUGA
4313
4143
ACAUCCUUUGUUCUCAUGA
4313
4161
UCAUGAGAACAAAGGAUGU
4571

4161
ACAGUAGGUCUGAGCAAAU
4314
4161
ACAGUAGGUCUGAGCAAAU
4314
4179
AUUUGCUCAGACCUACUGU
4572

4179
UGUUCCACCAAGCAUUUUC
4315
4179
UGUUCCACCAAGCAUUUUC
4315
4197
GAAAAUGCUUGGUGGAACA
4573

4197
CAGUGUCUUUGAAAAGCAC
4316
4197
CAGUGUCUUUGAAAAGCAC
4316
4215
GUGCUUUUCAAAGACACUG
4574

4215
CGUAACUUUUCAAAGGUGG
4317
4215
CGUAACUUUUCAAAGGUGG
4317
4233
CCACCUUUGAAAAGUUACG
4575

4233
GUCUUAAUUUGUUGCAUAU
4318
4233
GUCUUAAUUUGUUGCAUAU
4318
4251
AUAUGCAACAAAUUAAGAC
4576

4251
UCUAUCAAGGACUUAUUCA
4319
4251
UCUAUCAAGGACUUAUUCA
4319
4269
UGAAUAAGUCCUUGAUAGA
4577

4269
ACUCACCUUUCCUUUUCUG
4320
4269
ACUCACCUUUCCUUUUCUG
4320
4287
CAGAAAAGGAAAGGUGAGU
4578

4287
GCCCUCUAUCAAUUGAUUU
4321
4287
GCCCUCUAUCAAUUGAUUU
4321
4305
AAAUCAAUUGAUAGAGGGC
4579

4305
UCUUCUUACCUUUCAUCAU
4322
4305
UCUUCUUACCUUUCAUCAU
4322
4323
AUGAUGAAAGGUAAGAAGA
4580

4323
UUCAUUCCUUCCUUUAGAA
4323
4323
UUCAUUCCUUCCUUUAGAA
4323
4341
UUCUAAAGGAAGGAAUGAA
4581

4341
AAAACUGAAGAUUACCCAU
4324
4341
AAAACUGAAGAUUACCCAU
4324
4359
AUGGGUAAUCUUCAGUUUU
4582

4359
UAAUCUCCUCUUAUUACUU
4325
4359
UAAUCUCCUCUUAUUACUU
4325
4377
AAGUAAUAAGAGGAGAUUA
4583

4377
UGAGGGCCUUGACUAUUUA
4326
4377
UGAGGGCCUUGACUAUUUA
4326
4395
UAAAUAGUCAAGGCCCUCA
4584

4395
AGUUUAUUUUGUUUACUUU
4327
4395
AGUUUAUUUUGUUUACUUU
4327
4413
AAAGUAAACAAAAUAAACU
4585

4413
UACAGGUUAACACAGUUGU
4328
4413
UACAGGUUAACACAGUUGU
4328
4431
ACAACUGUGUUAACCUGUA
4586

4431
UUUUGUCUGAUUGCAUUUU
4329
4431
UUUUGUCUGAUUGCAUUUU
4329
4449
AAAAUGCAAUCAGACAAAA
4587

4449
UAUUAACUGUGAAGCCGUU
4330
4449
UAUUAACUGUGAAGCCGUU
4330
4467
AACGGCUUCACAGUUAAUA
4588

4467
UGAAAUGAAUAUCACUUAA
4331
4467
UGAAAUGAAUAUCACUUAA
4331
4485
UUAAGUGAUAUUCAUUUCA
4589

4485
AGCAACGUUGCUAAAUUUC
4332
4485
AGCAACGUUGCUAAAUUUC
4332
4503
GAAAUUUAGCAACGUUGCU
4590

4503
CUAUGUGUUUGAAAUGUGU
4333
4503
CUAUGUGUUUGAAAUGUGU
4333
4521
ACACAUUUCAAACACAUAG
4591

4521
UUAAUGAAGGCACUGCUUA
4334
4521
UUAAUGAAGGCACUGCUUA
4334
4539
UAAGCAGUGCCUUCAUUAA
4592

4539
AUUUGUAGUCACCUUGAAC
4335
4539
AUUUGUAGUCACCUUGAAC
4335
4557
GUUCAAGGUGACUACAAAU
4593

4557
CUGACUUAACCUAGAAGCU
4336
4557
CUGACUUAACCUAGAAGCU
4336
4575
AGCUUCUAGGUUAAGUCAG
4594

4575
UGUGCCUUCUUGUGAAAAA
4337
4575
UGUGCCUUCUUGUGAAAAA
4337
4593
UUUUUCACAAGAAGGCACA
4595

4593
AAAAAAAAAACAAAAACAA
4338
4593
AAAAAAAAAACAAAAACAA
4338
4611
UUGUUUUUGUUUUUUUUUU
4596

4611
AAAAACAGCCUUUAAACAA
4339
4611
AAAAACAGCCUUUAAACAA
4339
4629
UUGUUUAAAGGCUGUUUUU
4597

4629
AGUUUCCUUAGUGUCAAAA
4340
4629
AGUUUCCUUAGUGUCAAAA
4340
4647
UUUUGACACUAAGGAAACU
4598

HDAC11: NM_024827.1

3
CUUUGGGAGGGCCGGCCCC
4711
3
CUUUGGGAGGGCCGGCCCC
4711
21
GGGGCCGGCCCUCCCAAAG
4808

21
CGGGAUGCUACACACAACC
4712
21
CGGGAUGCUACACACAACC
4712
39
GGUUGUGUGUAGCAUCCCG
4809

39
CCAGGUGUACCAGGAUGUG
4713
39
CCAGCUGUACCAGCAUGUG
4713
57
CACAUGCUGGUACAGCUGG
4810

57
GCCAGAGACACCCUGGCCA
4714
57
GCCAGAGACACCCUGGCCA
4714
75
UGGCCAGGGUGUCUCUGGC
4811

75
AAUCGUGUACUCGCCGCGC
4715
75
AAUCGUGUACUCGCCGCGC
4715
93
GCGCGGCGAGUACACGAUU
4812

93
CUACAACAUCACCUUCAUG
4716
93
CUACAACAUCACCUUCAUG
4716
111
CAUGAAGGUGAUGUUGUAG
4813

111
GGGCCUGGAGAAGCUGCAU
4717
111
GGGCCUGGAGAAGCUGCAU
4717
129
AUGCAGCUUCUCCAGGCCC
4814

129
UCCCUUUGAUGCCGGAAAA
4718
129
UCCCUUUGAUGCCGGAAAA
4718
147
UUUUCCGGCAUCAAAGGGA
4815

147
AUGGGGCAAAGUGAUCAAU
4719
147
AUGGGGCAAAGUGAUCAAU
4719
165
AUUGAUCACUUUGCCCCAU
4816

165
UUUCCUAAAAGAAGAGAAG
4720
165
UUUCCUAAAAGAAGAGAAG
4720
183
CUUCUCUUCUUUUAGGAAA
4817

183
GCUUCUGUCUGACAGCAUG
4721
183
GCUUCUGUCUGACAGCAUG
4721
201
CAUGCUGUCAGACAGAAGC
4818

201
GCUGGUGGAGGCGCGGGAG
4722
201
GCUGGUGGAGGCGCGGGAG
4722
219
CUCCCGCGCCUCCACCAGC
4819

219
GGCCUCGGAGGAGGACCUG
4723
219
GGCCUCGGAGGAGGACCUG
4723
237
CAGGUCCUCCUCCGAGGCC
4820

237
GCUGGUGGUGCACACGAGG
4724
237
GCUGGUGGUGCACACGAGG
4724
255
CCUCGUGUGCACCACCAGC
4821

255
GCGCUAUCUUAAUGAGCUC
4725
255
GCGCUAUCUUAAUGAGCUC
4725
273
GAGCUCAUUAAGAUAGCGC
4822

273
CAAGUGGUCCUUUGCUGUU
4726
273
CAAGUGGUCCUUUGCUGUU
4726
291
AACAGCAAAGGACCACUUG
4823

291
UGCUACCAUCACAGAAAUC
4727
291
UGCUACCAUCACAGAAAUC
4727
309
GAUUUCUGUGAUGGUAGCA
4824

309
CCCCCCCGUUAUCUUCCUC
4728
309
CCCCCCCGUUAUCUUCCUC
4728
327
GAGGAAGAUAACGGGGGGG
4825

327
CCCCAACUUCCUUGUGCAG
4729
327
CCCCAACUUCCUUGUGCAG
4729
345
CUGCACAAGGAAGUUGGGG
4826

345
GAGGAAGGUGCUGAGGCCC
4730
345
GAGGAAGGUGCUGAGGCCC
4730
363
GGGCCUCAGCACCUUCCUC
4827

363
CCUUCGGACCCAGACAGGA
4731
363
CCUUCGGACCCAGACAGGA
4731
381
UCCUGUCUGGGUCCGAAGG
4828

381
AGGAACCAUAAUGGCGGGG
4732
381
AGGAACCAUAAUGGCGGGG
4732
399
CCCCGCCAUUAUGGUUCCU
4829

399
GAAGCUGGCUGUGGAGCGA
4733
399
GAAGCUGGCUGUGGAGCGA
4733
417
UCGCUCCACAGCCAGCUUC
4830

417
AGGCUGGGCCAUCAACGUG
4734
417
AGGCUGGGCCAUCAACGUG
4734
435
CACGUUGAUGGCCCAGCCU
4831

435
GGGGGGUGGCUUCCACCAC
4735
435
GGGGGGUGGCUUCCACCAC
4735
453
GUGGUGGAAGCCACCCCCC
4832

453
CUGCUCCAGCGACCGUGGC
4736
453
CUGCUCCAGCGACCGUGGC
4736
471
GCCACGGUCGCUGGAGCAG
4833

471
CGGGGGCUUCUGUGCCUAU
4737
471
CGGGGGCUUCUGUGCCUAU
4737
489
AUAGGCACAGAAGCCCCCG
4834

489
UGCGGACAUCACGCUCGCC
4738
489
UGCGGACAUCACGCUCGCC
4738
507
GGCGAGCGUGAUGUCCGCA
4835

507
CAUCAAGUUUCUGUUUGAG
4739
507
CAUCAAGUUUCUGUUUGAG
4739
525
CUCAAACAGAAACUUGAUG
4836

525
GCGUGUGGAGGGCAUCUCC
4740
525
GCGUGUGGAGGGCAUCUCC
4740
543
GGAGAUGCCCUCCACACGC
4837

543
CAGGGCUACCAUCAUUGAU
4741
543
CAGGGCUACCAUCAUUGAU
4741
561
AUCAAUGAUGGUAGCCCUG
4838

561
UCUUGAUGCCCAUCAGGGC
4742
561
UCUUGAUGCCCAUCAGGGC
4742
579
GCCCUGAUGGGCAUCAAGA
4839

579
CAAUGGGCAUGAGCGAGAC
4743
579
CAAUGGGCAUGAGCGAGAC
4743
597
GUCUCGCUCAUGCCCAUUG
4840

597
CUUCAUGGACGACAAGCGU
4744
597
CUUCAUGGACGACAAGCGU
4744
615
ACGCUUGUCGUCCAUGAAG
4841

615
UGUGUACAUCAUGGAUGUC
4745
615
UGUGUACAUCAUGGAUGUC
4745
633
GACAUCCAUGAUGUACACA
4842

633
CUACAACCGCCACAUCUAC
4746
633
CUACAACCGCCACAUCUAC
4746
651
GUAGAUGUGGCGGUUGUAG
4843

651
CCCAGGGGACCGCUUUGCC
4747
651
CCCAGGGGACCGCUUUGCC
4747
669
GGCAAAGCGGUCCCCUGGG
4844

669
CAAGCAGGCCAUCAGGCGG
4748
669
CAAGCAGGCCAUCAGGCGG
4748
687
CCGCCUGAUGGCCUGCUUG
4845

687
GAAGGUGGAGCUGGAGUGG
4749
687
GAAGGUGGAGCUGGAGUGG
4749
705
CCACUCCAGCUCCACCUUC
4846

705
GGGCACAGAGGAUGAUGAG
4750
705
GGGCACAGAGGAUGAUGAG
4750
723
CUCAUCAUCCUCUGUGCCC
4847

723
GUACCUGGAUAAGGUGGAG
4751
723
GUACCUGGAUAAGGUGGAG
4751
741
CUCCACCUUAUCCAGGUAC
4848

741
GAGGAACAUCAAGAAAUCC
4752
741
GAGGAACAUCAAGAAAUCC
4752
759
GGAUUUCUUGAUGUUCCUC
4849

759
CCUCCAGGAGCACCUGCCC
4753
759
CCUCCAGGAGCACCUGCCC
4753
777
GGGCAGGUGCUCCUGGAGG
4850

777
CGACGUGGUGGUAUACAAU
4754
777
CGACGUGGUGGUAUACAAU
4754
795
AUUGUAUACCACCACGUCG
4851

795
UGCAGGCACCGACAUCCUC
4755
795
UGCAGGCACCGACAUCCUC
4755
813
GAGGAUGUCGGUGCCUGCA
4852

813
CGAGGGGGACCGCCUUGGG
4756
813
CGAGGGGGACCGCCUUGGG
4756
831
CCCAAGGCGGUCCCCCUCG
4853

831
GGGGCUGUCCAUCAGCCCA
4757
831
GGGGCUGUCCAUCAGCCCA
4757
849
UGGGCUGAUGGACAGCCCC
4854

849
AGCGGGCAUCGUGAAGCGG
4758
849
AGCGGGCAUCGUGAAGCGG
4758
867
CCGCUUCACGAUGCCCGCU
4855

867
GGAUGAGCUGGUGUUCCGG
4759
867
GGAUGAGCUGGUGUUCCGG
4759
885
CCGGAACACCAGCUCAUCC
4856

885
GAUGGUCCGUGGCCGCCGG
4760
885
GAUGGUCCGUGGCCGCCGG
4760
903
CCGGCGGCCACGGACCAUC
4857

903
GGUGCCCAUCCUUAUGGUG
4761
903
GGUGCCCAUCCUUAUGGUG
4761
921
CACCAUAAGGAUGGGCACC
4858

921
GACCUCAGGCGGGUACCAG
4762
921
GACCUCAGGCGGGUACCAG
4762
939
CUGGUACCCGCCUGAGGUC
4859

939
GAAGCGCACAGCCCGCAUC
4763
939
GAAGCGCACAGCCCGCAUC
4763
957
GAUGCGGGCUGUGCGCUUC
4860

957
CAUUGCUGACUCCAUACUU
4764
957
CAUUGCUGACUCCAUACUU
4764
975
AAGUAUGGAGUCAGCAAUG
4861

975
UAAUCUGUUUGGCCUGGGG
4765
975
UAAUCUGUUUGGCCUGGGG
4765
993
CCCCAGGCCAAACAGAUUA
4862

993
GCUCAUUGGGCCUGAGUCA
4766
993
GCUCAUUGGGCCUGAGUCA
4766
1011
UGACUCAGGCCCAAUGAGC
4863

1011
ACCCAGCGUCUCCGCACAG
4767
1011
ACCCAGCGUCUCCGCACAG
4767
1029
CUGUGCGGAGACGCUGGGU
4864

1029
GAACUCAGACACACCGCUG
4768
1029
GAACUCAGACACACCGCUG
4768
1047
CAGCGGUGUGUCUGAGUUC
4865

1047
GCUUCCCCCUGCAGUGCCC
4769
1047
GCUUCCCCCUGCAGUGCCC
4769
1065
GGGCACUGCAGGGGGAAGC
4866

1065
CUGACCCUUGCUGCCCUGC
4770
1065
CUGACCCUUGCUGCCCUGC
4770
1083
GCAGGGCAGCAAGGGUCAG
4867

1083
CCUGUCACGUGGCCCUGCC
4771
1083
CCUGUCACGUGGCCCUGCC
4771
1101
GGCAGGGCCACGUGACAGG
4868

1101
CUAUCCGCCCCUUAGUGCU
4772
1101
CUAUCCGCCCCUUAGUGCU
4772
1119
AGCACUAAGGGGCGGAUAG
4869

1119
UUUUUGUUUUCUAACCUCA
4773
1119
UUUUUGUUUUCUAACCUCA
4773
1137
UGAGGUUAGAAAACAAAAA
4870

1137
AUGGGGUGGUGGAGGCAGC
4774
1137
AUGGGGUGGUGGAGGCAGC
4774
1155
GCUGCCUCCACCACCCCAU
4871

1155
CCUUCAGUGAGCAUGGAGG
4775
1155
CCUUCAGUGAGCAUGGAGG
4775
1173
CCUCCAUGCUCACUGAAGG
4872

1173
GGGCAGGGCCAUCCCUGGC
4776
1173
GGGCAGGGCCAUCCCUGGC
4776
1191
GCCAGGGAUGGCCCUGCCC
4873

1191
CUGGGGCCUGGAGCUGGCC
4777
1191
CUGGGGCCUGGAGCUGGCC
4777
1209
GGCCAGCUCCAGGCCCCAG
4874

1209
CCUUCCUCUACUUUUCCCU
4778
1209
CCUUCCUCUACUUUUCCCU
4778
1227
AGGGAAAAGUAGAGGAAGG
4875

1227
UGCUGGAAGCCAGAAGGGC
4779
1227
UGCUGGAAGCCAGAAGGGC
4779
1245
GCCCUUCUGGCUUCCAGCA
4876

1245
CUUGAGGCCUCUAUGGGUG
4780
1245
CUUGAGGCCUCUAUGGGUG
4780
1263
CACCCAUAGAGGCCUCAAG
4877

1263
GGGGGCAGAAGGCAGAGCC
4781
1263
GGGGGCAGAAGGCAGAGCC
4781
1281
GGCUCUGCCUUCUGCCCCC
4878

1281
CUGUGUCCCAGGGGGACCC
4782
1281
CUGUGUCCCAGGGGGACCC
4782
1299
GGGUCCCCCUGGGACACAG
4879

1299
CACACGAAGUCACCAGCCC
4783
1299
CACACGAAGUCACCAGCCC
4783
1317
GGGCUGGUGACUUCGUGUG
4880

1317
CAUAGGUCCAGGGAGGCAG
4784
1317
CAUAGGUCCAGGGAGGCAG
4784
1335
CUGCCUCCCUGGACCUAUG
4881

1335
GGCAGUUAACUGAGAAUUG
4785
1335
GGCAGUUAACUGAGAAUUG
4785
1353
CAAUUCUCAGUUAACUGCC
4882

1353
GGAGAGGACAGGCUAGGUC
4786
1353
GGAGAGGACAGGCUAGGUC
4786
1371
GACCUAGCCUGUCCUCUCC
4883

1371
CCCAGGCACAGCGAGGGCC
4787
1371
CCCAGGCACAGCGAGGGCC
4787
1389
GGCCCUCGCUGUGCCUGGG
4884

1389
CCUGGGCUUGGGGUGUUCU
4788
1389
CCUGGGCUUGGGGUGUUCU
4788
1407
AGAACACCCCAAGCCCAGG
4885

1407
UGGUUUUGAGAACGGCAGA
4789
1407
UGGUUUUGAGAACGGCAGA
4789
1425
UCUGCCGUUCUCAAAACCA
4886

1425
ACCCAGGUCGGAGUGAGGA
4790
1425
ACCCAGGUCGGAGUGAGGA
4790
1443
UCCUCACUCCGACCUGGGU
4887

1443
AAGCUUCCACCUCCAUCCU
4791
1443
AAGCUUCCACCUCCAUCCU
4791
1461
AGGAUGGAGGUGGAAGCUU
4888

1461
UGACUAGGCCUGCAUCCUA
4792
1461
UGACUAGGCCUGCAUCCUA
4792
1479
UAGGAUGCAGGCCUAGUCA
4889

1479
AACUGGGCCUCCCUCCCUC
4793
1479
AACUGGGCCUCCCUCCCUC
4793
1497
GAGGGAGGGAGGCCCAGUU
4890

1497
CCCCUUGGUCAUGGGAUUU
4794
1497
CCCCUUGGUCAUGGGAUUU
4794
1515
AAAUCCCAUGACCAAGGGG
4891

1515
UGCUGCCCUCUUUGCCCCA
4795
1515
UGCUGCCCUCUUUGCCCCA
4795
1533
UGGGGCAAAGAGGGCAGCA
4892

1533
AGAGCUGAAGAGCUAUAGG
4796
1533
AGAGCUGAAGAGCUAUAGG
4796
1551
CCUAUAGCUCUUCAGCUCU
4893

1551
GCACUGGUGUGGAUGGCCC
4797
1551
GCACUGGUGUGGAUGGCCC
4797
1569
GGGCCAUCCACACCAGUGC
4894

1569
CAGGAGGUGCUGGAGCUAG
4798
1569
CAGGAGGUGGUGGAGGUAG
4798
1587
CUAGCUCCAGCACCUCCUG
4895

1587
GGUCUCCAGGUGGGCCUGG
4799
1587
GGUCUCCAGGUGGGCCUGG
4799
1605
CCAGGCCCACCUGGAGACC
4896

1605
GUUCCCAGGCAGCAGGUGG
4800
1605
GUUCCCAGGCAGCAGGUGG
4800
1623
CCACCUGCUGCCUGGGAAC
4897

1623
GGAACCCUGGGCCUGGAUG
4801
1623
GGAACCCUGGGCCUGGAUG
4801
1641
CAUCCAGGCCCAGGGUUCC
4898

1641
GUGAGGGGCGGUCAGGAAG
4802
1641
GUGAGGGGCGGUCAGGAAG
4802
1659
CUUCCUGACCGCCCCUCAC
4899

1659
GGGGUACAGGUGGGUUCCC
4803
1659
GGGGUACAGGUGGGUUCCC
4803
1677
GGGAACCCACCUGUACCCC
4900

1677
CUCAUGUGGAGUUCCCCCU
4804
1677
CUCAUCUGGAGUUCCCCCU
4804
1695
AGGGGGAACUCCAGAUGAG
4901

1695
UCAAUAAAGCAAGGUCUGG
4805
1695
UCAAUAAAGCAAGGUCUGG
4805
1713
CCAGACCUUGCUUUAUUGA
4902

1713
GACCUGCAAAAAAAAAAAA
4806
1713
GACCUGCAAAAAAAAAAAA
4806
1731
UUUUUUUUUUUUGCAGGUC
4903

1731
AAAAAAAAAAAAAAAAAAA
4807
1731
AAAAAAAAAAAAAAAAAAA
4807
1749
UUUUUUUUUUUUUUUUUUU
4904

The 3′-ends of the Upper sequence and the Lower sequence of the siNA construct can include an overhang sequence, for example about 1, 2, 3, or 4 nucleotides in length, preferably 2 nucleotides in length, wherein the overhanging sequence of the lower sequence is optionally complementary to a portion of the target sequence. The upper sequence is also referred to as the sense strand, whereas the lower sequence is also referred to as the antisense strand. The upper and lower sequences in the Table can further comprise a chemical modification having Formulae I-VII, such as exemplary siNA constructs shown in FIGS. 4 and 5, or having modifications described in Table IV or any combination thereof.

TABLE IIIHDAC synthetic siNA and Target SequencesTar-getSeqSeqPosTargetIDCmpd#AliasesSequenceIDHDAC1744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 744U21 siNA senseGCUCCGAGACGGGAUUGAUTT2391892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1892U21 siNA senseGGCUCCUAAAGUAACAUCATT2401921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1921U21 siNA senseGAUUGGUUCUGUUUUCGUATT241743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 743U21 siNA senseCGCUCCGAGACGGGAUUGATT242963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 963U21 siNA senseCGGUGGUUACACCAUUCGUTT2431717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1717U21 siNA senseCGUUCUUAACUUUGAACCATT24430CGGACCGACUGACGGUAGGGACG237HDAC1: 30U21 siNA senseGACCGACUGACGGUAGGGATT245741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 741U21 siNA senseCCCGCUCCGAGACGGGAUUTT246744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 762L21 siNA antisenseAUCAAUCCCGUCUCGGAGCTT247(744C)1892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1910L21 siNA antisenseUGAUGUUACUUUAGGAGCCTT248(1892C)1921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1939L21 siNA antisenseUACGAAAACAGAACCAAUCTT249(1921C)743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 761L21 siNA antisenseUCAAUCCCGUCUCGGAGCGTT250(743C)963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 981L21 siNA antisenseACGAAUGGUGUAACCACCGTT251(963C)1717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1735L21 siNA antisenseUGGUUCAAAGUUAAGAACGTT252(1717C)30CGGACCGACUGACGGUAGGGACG237HDAC1: 48L21 siNA antisenseUCCCUACCGUCAGUCGGUCTT253(30C)741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 759L21 siNA antisenseAAUCCCGUCUCGGAGCGGGTT254(741C)744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 744U21 siNA sense stab04B GcuccGAGAcGGGAuuGAuTT B2551892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1892U21 siNA senseB GGcuccuAAAGuAAcAucATT B256stab041921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1921U21 siNA senseB GAuuGGuucuGuuuucGuATT B257stab04743CGCGCUCCGAGACGGGAUUGAUG234HDAC1: 743U21 siNA sense stab04B cGcuccGAGAcGGGAuuGATT B258963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 963U21 siNA sense stab04B cGGuGGuuAcAccAuucGuTT B2591717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1717U21 siNA senseB cGuucuuAAcuuuGAAccATT B260stab0430CGGACCGACUGACGGUAGGGACG237HDAC1: 30U21 siNA sense stab04B GAccGAcuGAcGGuAGGGATT B261741UACCGGCUCCGAGACGGGAUUGA238HDAC1: 741U21 siNA sense stab04B cccGcuccGAGAcGGGAuuTT B262744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 762L21 siNA antisenseAucAAucccGucucGGAGcTsT263(744C) stab051892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1910L21 siNA antisenseuGAuGuuAcuuuAGGAGccTsT264(1892C) stab051921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1939L21 siNA antisenseuAcGAAAAcAGAAccAAucTsT265(1921C) stab05743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 761L21 siNA antisenseucAAucccGucucGGAGcGTsT266(743C) stab05963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 981L21 siNA antisenseAcGAAuGGuGuAAccAccGTsT267(963C) stab051717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1735L21 siNA antisenseuGGuucAAAGuuAAGAAcGTsT268(1717C) stab0530CGGACCGACUGACGGUAGGGACG237HDAC1: 48L21 siNA antisenseucccuAccGucAGucGGucTsT269(30C) stab05741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 759L21 siNA antisenseAAucccGucucGGAGcGGGTsT270(741C) stab05744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 744U21 siNA sense stab07B GcuccGAGAcGGGAuuGAuTT B2711892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1892U21 siNA senseB GGcuccuAAAGuAAcAucATT B272stab071921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1921U21 siNA senseB GAuuGGuucuGuuuucGuATT B273stab07743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 743U21 siNA sense stab07B cGcuccGAGAcGGGAuuGATT B274963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 963U21 siNA sense stab07B cGGuGGuuAcAccAuucGuTT B2751717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1717U21 siNA senseB cGuucuuAAcuuuGAAccATT B276stab0730CGGACCGACUGACGGUAGGGACG237HDAC1: 30U21 siNA sense stab07B GAccGAcuGAcGGuAGGGATT B277741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 741U21 siNA sense stab07B cccGcuccGAGAcGGGAuuTT B278744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 762L21 siNA antisenseAucAAucccGucucGGAGcTsT279(744C) stab111892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1910L21 siNA antisenseuGAuGuuAcuuuAGGAGccTsT280(1892C) stab111921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1939L21 siNA antisenseuAcGAAAAcAGAAccAAucTsT281(1921C) stab11743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 761L21 siNA antisenseucAAucccGucucGGAGcGTsT282(743C) stab11963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 981L21 siNA antisenseAcGAAuGGuGuAAccAccGTsT283(963C) stab111717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1735L21 siNA antisenseuGGuucAAAGuuAAGAAcGTsT284(1717C) stab1130CGGACCGACUGACGGUAGGGACG237HDAC1: 48L21 siNA antisenseucccuAccGucAGucGGucTsT285(30C) stab11741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 759L21 siNA antisenseAAucccGucucGGAGcGGGTsT286(741C) stab11744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 744U21 siNA sense stab18B GcuccGAGAcGGGAuuGAuTT B2871892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1892U21 siNA senseB GGcuccuAAAGuAAcAucATT B288stab181921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1921U21 siNA senseB GAuuGGuucuGuuuucGuATT B289stab18743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 743U21 siNA sense stab18B cGcuccGAGAcGGGAuuGATT B290963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 963U21 siNA sense stab18B cGGuGGuuAcAccAuucGuTT B2911717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1717U21 siNA senseB cGuucuuAAcuuuGAAccATT B292stab1830CGGACCGACUGACGGUAGGGACG237HDAC1: 30U21 siNA sense stab18B GAccGAcuGAcGGuAGGGATT B293741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 741U21 siNA sense stab18B cccGcuccGAGAcGGGAuuTT B294744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 762L21 siNA antisenseAucAAucccGucucGGAGcTsT295(744C) stab081892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1910L21 siNA antisenseuGAuGuuAcuuuAGGAGccTsT296(1892C) stab081921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1939L21 siNA antisenseuAcGAAAAcAGAAccAAucTsT297(1921C) stab08743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 761L21 siNA antisenseucAAucccGucucGGAGcGTsT298(743C) stab08963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 981L21 siNA antisenseAcGAAuGGuGuAAccAccGTsT299(963C) stab081717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1735L21 siNA antisenseuGGuucAAAGuuAAGAAcGTsT300(1717C) stab0830CGGACCGACUGACGGUAGGGACG237HDAC1: 48L21 siNA antisenseucccuAccGucAGucGGucTsT301(30C) stab08741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 759L21 siNA antisenseAAucccGucucGGAGcGGGTsT302(741C) stab08744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 744U21 siNA sense stab09B GCUCCGAGACGGGAUUGAUTT B3031892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1892U21 siNA senseB GGCUCCUAAAGUAACAUCATT B304stab091921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1921U21 siNA senseB GAUUGGUUCUGUUUUCGUATT B305stab09743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 743U21 siNA sense stab09B CGCUCCGAGACGGGAUUGATT B306963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 963U21 siNA sense stab09B CGGUGGUUACACCAUUCGUTT B3071717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1717U21 siNA senseB CGUUCUUAACUUUGAACCATT B308stab0930CGGACCGACUGACGGUAGGGACG237HDAC1: 30U21 siNA sense stab09B GACCGACUGACGGUAGGGATT B309741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 741U21 siNA sense stab09B CCCGCUCCGAGACGGGAUUTT B310744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 762L21 siNA antisenseAUCAAUCCCGUCUCGGAGCTsT311(744C) stab101892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1910L21 siNA antisenseUGAUGUUACUUUAGGAGCCTsT312(1892C) stab101921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1939L21 siNA antisenseUACGAAAACAGAACCAAUCTsT313(1921C) stab10743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 761L21 siNA antisenseUCAAUCCCGUCUCGGAGCGTsT314(743C) stab10963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 981L21 siNA antisenseACGAAUGGUGUMCCACCGTsT315(963C) stab101717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1735L21 siNA antisenseUGGUUCAAAGUUAAGAACGTsT316(1717C) stab1030CGGACCGACUGACGGUAGGGACG237HDAC1: 48L21 siNA antisenseUCCCUACCGUCAGUCGGUCTsT317(30C) stab10741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 759L21 siNA antisenseAAUCCCGUCUCGGAGCGGGTsT318(741C) stab10744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 762L21 siNA antisenseAucAAucccGucucGGAGcTT B319(744C) stab191892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1910L21 siNA antisenseuGAuGuuAcuuuAGGAGccTT B320(1892C) stab191921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1939L21 siNA antisenseuAcGAAAAcAGAAccAAucTT B321(1921C) stab19743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 761L21 siNA antisenseucAAucccGucucGGAGcGTT B322(743C) stab19963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 981L21 siNA antisenseAcGAAuGGuGuAAccAccGTT B323(963C) stab191717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1735L21 siNA antisenseuGGuucAAAGuuAAGAAcGTT B324(1717C) stab1930CGGACCGACUGACGGUAGGGACG237HDAC1: 48L21 siNA antisenseucccuAccGucAGucGGucTT B325(30C) stab19741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 759L21 siNA antisenseAAucccGucucGGAGcGGGTT B326(741C) stab19744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 762L21 siNA antisenseAUCAAUCCCGUCUCGGAGCTT B327(744C) stab221892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1910L21 siNA antisenseUGAUGUUACUUUAGGAGCCTT B328(1892C) stab221921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1939L21 siNA antisenseUACGAAAACAGAACCAAUCTT B329(1921C) stab22743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 761L21 siNA antisenseUCAAUCCCGUCUCGGAGCGTT B330(743C) stab22963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 981L21 siNA antisenseACGAAUGGUGUAACCACCGTT B331(963C) stab221717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1735L21 siNA antisenseUGGUUCAAAGUUAAGAACGTT B332(1717C) stab2230CGGACCGACUGACGGUAGGGACG237HDAC1: 48L21 siNA antisenseUCCCUACCGUCAGUCGGUCTT B333(30C) stab22741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 759L21 siNA antisenseAAUCCCGUCUCGGAGCGGGTT B334(741C) stab22744CCGCUCCGAGACGGGAUUGAUGA231HDAC1: 762L21 siNA antisenseAUCAAucccGucucGGAGcTsT335(744C) stab251892CAGGCUCCUAAAGUAACAUCAGC232HDAC1: 1910L21 siNA antisenseUGAuGuuAcuuuAGGAGccTsT336(1892C) stab251921UAGAUUGGUUCUGUUUUCGUACC233HDAC1: 1939L21 siNA antisenseUACGAAAAcAGAAccAAucTsT337(1921C) stab25743CCCGCUCCGAGACGGGAUUGAUG234HDAC1: 761L21 siNA antisenseUCAAucccGucucGGAGcGTsT338(743C) stab25963GGCGGUGGUUACACCAUUCGUAA235HDAC1: 981L21 siNA antisenseACGAAuGGuGuAAccAccGTsT339(963C) stab251717CCCGUUCUUAACUUUGAACCAUA236HDAC1: 1735L21 siNA antisenseUGGuucAAAGuuAAGAAcGTsT340(1717C) stab2530CGGACCGACUGACGGUAGGGACG237HDAC1: 48L21 siNA antisenseUCCcuAccGucAGucGGucTsT341(30C) stab25741UACCCGCUCCGAGACGGGAUUGA238HDAC1: 759L21 siNA antisenseAAUcccGucucGGAGcGGGTsT342(741C) stab25HDAC2223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 223U21 siNA senseGGCGGCAAAAAAAAAGUCUTT571543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 543U21 siNA senseCUCAACUGGCGGUUCAGUUTT572781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 781U21 siNA senseCGUGUAAUGACGGUAUCAUTT573782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 782U21 siNA senseGUGUAAUGACGGUAUCAUUTT5741012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1012U21 siNA senseGAUAGACUGGGUUGUUUCATT575957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 957U21 siNA senseGAUGUAUCAACCUAGUGCUTT576985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 985U21 siNA senseCAGUGUGGUGCAGACUCAUTT577776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 776U21 siNA senseCAGAUCGUGUAAUGACGGUTT578223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 241L21 siNA antisenseAGACUUUUUUUUUGCCGCCTT579(223C)543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 561L21 siNA antisenseAACUGAACCGCCAGUUGAGTT580(543C)781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 799L21 siNA antisenseAUGAUACCGUCAUUACACGTT581(781C)782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 800L21 siNA antisenseAAUGAUACCGUCAUUACACTT582(782C)1012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1030L21 siNA antisenseUGAAACAACCCAGUCUAUCTT583(1012C)957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 975L21 siNA antisenseAGCACUAGGUUGAUACAUCTT584(957C)985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 1003L21 siNA antisenseAUGAGUCUGCACCACACUGTT585(985C)776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 794L21 siNA antisenseACCGUCAUUACACGAUCUGTT586(776C)223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 223U21 siNA sense stab04B GGcGGcAAAAAAAAAGucuTT B587543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 543U21 siNA sense stab04B cucAAcuGGcGGuucAGuuTT B588781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 781U21 siNA sense stab04B cGuGuAAuGAcGGuAucAuTT B589782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 782U21 siNA sense stab04B GuGuAAuGAcGGuAucAuuTT B5901012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1012U21 siNA senseB GAuAGAcuGGGuuGuuucATT B591stab04957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 957U21 siNA sense stab04B GAuGuAucAAccuAGuGcuTT B592985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 985U21 siNA sense stab04B cAGuGuGGuGcAGAcucAuTT B593776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 776U21 siNA sense stab04B cAGAucGuGuAAuGAcGGuTT B594223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 241L21 siNA antisenseAGAcuuuuuuuuuGccGccTsT595(223C) stab05543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 561L21 siNA antisenseAAcuGAAccGccAGuuGAGTsT596(543C) stab05781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 799L21 siNA antisenseAuGAuAccGucAuuAcAcGTsT597(781C) stab05782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 800L21 siNA antisenseAAuGAuAccGucAuuAcAcTsT598(782C) stab051012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1030L21 siNA antisenseuGAAAcAAcccAGucuAucTsT599(1012C) stab05957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 975L21 siNA antisenseAGcAcuAGGuuGAuAcAucTsT600(957C) stab05985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 1003L21 siNA antisenseAuGAGucuGcAccAcAcuGTsT601(985C) stab05776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 794L21 siNA antisenseAccGucAuuAcAcGAucuGTsT602(776C) stab05223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 223U21 siNA sense stab07B GGcGGcAAAAAAAAAGucuTT B603543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 543U21 siNA sense stab07B cucAAcuGGcGGuucAGuuTT B604781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 781U21 siNA sense stab07B cGuGuAAuGAcGGuAucAuTT B605782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 782U21 siNA sense stab07B GuGuAAuGAcGGuAucAuuTT B6061012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1012U21 siNA senseB GAuAGAcuGGGuuGuuucATT B607stab07957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 957U21 siNA sense stab07B GAuGuAucAAccuAGuGcuTT B608985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 985U21 siNA sense stab07B cAGuGuGGuGcAGAcucAuTT B609776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 776U21 siNA sense stab07B cAGAucGuGuAAuGAcGGuTT B610223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 241L21 siNA antisenseAGAcuuuuuuuuuGccGccTsT611(223C) stab11543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 561L21 siNA antisenseAAcuGAAccGccAGuuGAGTsT612(543C) stab11781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 799L21 siNA antisenseAuGAuAccGucAuuAcAcGTsT613(781C) stab11782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 800L21 siNA antisenseAAuGAuAccGucAuuAcAcTsT614(782C) stab111012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1030L21 siNA antisenseuGAAAcAAcccAGucuAucTsT615(1012C) stab11957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 975L21 siNA antisenseAGcAcuAGGuuGAuAcAucTsT616(957C) stab11985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 1003L21 siNA antisenseAuGAGucuGcAccAcAcuGTsT617(985C) stab11776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 794L21 siNA antisenseAccGucAuuAcAcGAucuGTsT618(776C) stab11223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 223U21 siNA sense stab18B GGcGGcAAAAAAAAAGucuTT B619543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 543U21 siNA sense stab18B cucAAcuGGcGGuucAGuuTT B620781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 781U21 siNA sense stab18B cGuGuAAuGAcGGuAucAuTT B621782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 782U21 siNA sense stab18B GuGuAAuGAcGGuAucAuuTT B6221012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1012U21 siNA senseB GAuAGAcuGGGuuGuuucATT B623stab18957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 957U21 siNA sense stab18B GAuGuAucAAccuAGuGcuTT B624985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 985U21 siNA sense stab18B cAGuGuGGuGcAGAcucAuTT B625776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 776U21 siNA sense stab18B cAGAucGuGuAAuGAcGGuTT B626223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 241L21 siNA antisenseAGAcuuuuuuuuuGccGccTsT627(223C) stab08543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 561L21 siNA antisenseAAcuGAAccGccAGuuGAGTsT628(543C) stab08781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 799L21 siNA antisenseAuGAuAccGucAuuAcAcGTsT629(781C) stab08782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 800L21 siNA antisenseAAuGAuAccGucAuuAcAcTsT630(782C) stab081012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1030L21 siNA antisenseuGAAAcAAcccAGucuAucTsT631(1012C) stab08957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 975L21 siNA antisenseAGcAcuAGGuuGAuAcAucTsT632(957C) stab08985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 1003L21 siNA antisenseAuGAGucuGcAccAcAcuGTsT633(985C) stab08776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 794L21 siNA antisenseAccGucAuuAcAcGAucuGTsT634(776C) stab08223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 223U21 siNA sense stab09B GGCGGCAAAAAAAAAGUCUTT B635543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 543U21 siNA sense stab09B CUCAACUGGCGGUUCAGUUTT B636781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 781U21 siNA sense stab09B CGUGUAAUGACGGUAUCAUTT B637782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 782U21 siNA sense stab09B GUGUAAUGACGGUAUCAUUTT B6381012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1012U21 siNA senseB GAUAGACUGGGUUGUUUCATT B639stab09957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 957U21 siNA sense stab09B GAUGUAUCAACCUAGUGCUTT B640985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 985U21 siNA sense stab09B CAGUGUGGUGCAGACUCAUTT B641776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 776U21 siNA sense stab09B CAGAUCGUGUAAUGACGGUTT B642223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 241L21 siNA antisenseAGACUUUUUUUUUGCCGCCTsT643(223C) stab10543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 561L21 siNA antisenseAACUGAACCGCCAGUUGAGTsT644(543C) stab10781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 799L21 siNA antisenseAUGAUACCGUCAUUACACGTsT645(781C) stab10782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 800L21 siNA antisenseAAUGAUACCGUCAUUACACTsT646(782C) stab101012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1030L21 siNA antisenseUGAAACAACCCAGUCUAUCTsT647(1012C) stab10957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 975L21 siNA antisenseAGCACUAGGUUGAUACAUCTsT648(957C) stab10985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 1003L21 siNA antisenseAUGAGUCUGCACCACACUGTsT649(985C) stab10776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 794L21 siNA antisenseACCGUCAUUACACGAUCUGTsT650(776C) stab10223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 241L21 siNA antisenseAGAcuuuuuuuuuGccGccTT B651(223C) stab19543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 561L21 siNA antisenseAAcuGAAccGccAGuuGAGTT B652(543C) stab19781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 799L21 siNA antisenseAuGAuAccGucAuuAcAcGTT B653(781C) stab19782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 800L21 siNA antisenseAAuGAuAccGucAuuAcAcTT B654(782C) stab191012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1030L21 siNA antisenseuGAAAcAAcccAGucuAucTT B655(1012C) stab19957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 975L21 siNA antisenseAGcAcuAGGuuGAuAcAucTT B656(957C) stab19985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 1003L21 siNA antisenseAuGAGucuGcAccAcAcuGTT B657(985C) stab19776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 794L21 siNA antisenseAccGucAuuAcAcGAucuGTT B658(776C) stab19223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 241L21 siNA antisenseAGACUUUUUUUUUGCCGCCTT B659(223C) stab22543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 561L21 siNA antisenseAACUGAACCGCCAGUUGAGTT B660(543C) stab22781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 799L21 siNA antisenseAUGAUACCGUCAUUACACGTT B661(781C) stab22782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 800L21 siNA antisenseAAUGAUACCGUCAUUACACTT B662(782C) stab221012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1030L21 siNA antisenseUGAAACAACCCAGUCUAUCTT B663(1012C) stab22957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 975L21 siNA antisenseAGCACUAGGUUGAUACAUCTT B664(957C) stab22985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 1003L21 siNA antisenseAUGAGUCUGCACCACACUGTT B665(985C) stab22776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 794L21 siNA antisenseACCGUCAUUACACGAUCUGTT B666(776C) stab22223GAGGCGGCAAAAAAAAAGUCUGC563HDAC2: 241L21 siNA antisenseAGAcuuuuuuuuuGccGccTsT667(223C) stab25543CUCUCAACUGGCGGUUCAGUUGC564HDAC2: 561L21 siNA antisenseAACuGAAccGccAGuuGAGTsT668(543C) stab25781AUCGUGUAAUGACGGUAUCAUUC565HDAC2: 799L21 siNA antisenseAUGAuAccGucAuuAcAcGTsT669(781C) stab25782UCGUGUAAUGACGGUAUCAUUCC566HDAC2: 800L21 siNA antisenseAAUGAuAccGucAuuAcAcTsT670(782C) stab251012GUGAUAGACUGGGUUGUUUCAAU567HDAC2: 1030L21 siNA antisenseUGAAAcAAcccAGucuAucTsT671(1012C) stab25957GAGAUGUAUCAACCUAGUGCUGU568HDAC2: 975L21 siNA antisenseAGCAcuAGGuuGAuAcAucTsT672(957C) stab25985UACAGUGUGGUGCAGACUCAUUA569HDAC2: 1003L21 siNA antisenseAUGAGucuGcAccAcAcuGTsT673(985C) stab25776AACAGAUCGUGUAAUGACGGUAU570HDAC2: 794L21 siNA antisenseACCGucAuuAcAcGAucuGTsT674(776C) stab25HDAC3361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 361U21 siNA senseGUUCUGCUCGCGUUACACATT899849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 849U21 siNA senseGAUUGGGCUGCUUUAACCUTT900768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 768U21 siNA senseCGGUUAUCAACCAGGUAGUTT901781CAGGUAGUGGACUUCUACCAACC894HDAC3: 781U21 siNA senseGGUAGUGGACUUCUACCAATT9021484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1484U21 siNA senseGUUCUCGAACCAUCUACCUTT9031538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1538U21 siNA senseCCUAUUAGGGAUGGAGAUATT904315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 315U21 siNA senseCCUUCAACGUAGGCGAUGATT905355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 355U21 siNA senseCUUUGAGUUCUGCUCGCGUTT906361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 379L21 siNA antisenseUGUGUAACGCGAGCAGAACTT907(361C)849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 867L21 siNA antisenseAGGUUAAAGCAGCCCAAUCTT908(849C)768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 786L21 siNA antisenseACUACCUGGUUGAUAACCGTT909(768C)781CAGGUAGUGGACUUCUACCAACC894HDAC3: 799L21 siNA antisenseUUGGUAGAAGUCCACUACCTT910(781C)1484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1502L21 siNA antisenseAGGUAGAUGGUUCGAGAACTT911(1484C)1538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1556L21 siNA antisenseUAUCUCCAUCCCUAAUAGGTT912(1538C)315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 333L21 siNA antisenseUCAUCGCCUACGUUGAAGGTT913(315C)355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 373L21 siNA antisenseACGCGAGCAGAACUCAAAGTT914(355C)361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 361U21 siNA sense stab04B GuucuGcucGcGuuAcAcATT B915849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 849U21 siNA sense stab04B GAuuGGGcuGcuuuAAccuTT B916768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 768U21 siNA sense stab04B cGGuuAucAAccAGGuAGuTT B917781CAGGUAGUGGACUUCUACCAACC894HDAC3: 781U21 siNA sense stab04B GGuAGuGGAcuucuAccAATT B9181484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1484U21 siNA senseB GuucucGAAccAucuAccuTT B919stab041538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1538U21 siNA senseB ccuAuuAGGGAuGGAGAuATT B920stab04315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 315U21 siNA sense stab04B ccuucAAcGuAGGcGAuGATT B921355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 355U21 siNA sense stab04B cuuuGAGuucuGcucGcGuTT B922361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 379L21 siNA antisenseuGuGuAAcGcGAGcAGAAcTsT923(361C) stab05849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 867L21 siNA antisenseAGGuuAAAGcAGcccAAucTsT924(849C) stab05768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 786L21 siNA antisenseAcuAccuGGuuGAuAAccGTsT925(768C) stab05781CAGGUAGUGGACUUCUACCAACC894HDAC3: 799L21 siNA antisenseuuGGuAGAAGuccAcuAccTsT926(781C) stab051484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1502L21 siNA antisenseAGGuAGAuGGuucGAGAAcTsT927(1484C) stab051538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1556L21 siNA antisenseuAucuccAucccuAAuAGGTsT928(1538C) stab05315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 333L21 siNA antisenseucAucGccuAcGuuGAAGGTsT929(315C) stab05355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 373L21 siNA antisenseAcGcGAGcAGAAcucAAAGTsT930(355C) stab05361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 361U21 siNA sense stab07B GuucuGcucGcGuuAcAcATT B931849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 849U21 siNA sense stab07B GAuuGGGcuGcuuuAAccuTT B932768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 768U21 siNA sense stab07B cGGuuAucAAccAGGuAGuTT B933781CAGGUAGUGGACUUCUACCAACC894HDAC3: 781U21 siNA sense stab07B GGuAGuGGAcuucuAccAATT B9341484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1484U21 siNA senseB GuucucGAAccAucuAccuTT B935stab071538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1538U21 siNA senseB ccuAuuAGGGAuGGAGAuATT B936stab07315UGCCUUCAACGUAGGGGAUGACU897HDAC3: 315U21 siNA sense stab07B ccuucAAcGuAGGcGAuGATT B937355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 355U21 siNA sense stab07B cuuuGAGuucuGcucGcGuTT B938361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 379L21 siNA antisenseuGuGuAAcGcGAGcAGAAcTsT939(361C) stab11849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 867L21 siNA antisenseAGGuuAAAGcAGcccAAucTsT940(849C) stab11768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 786L21 siNA antisenseAcuAccuGGuuGAuAAccGTsT941(768C) stab11781CAGGUAGUGGACUUCUACCAACC894HDAC3:799L21 siNA antisenseuuGGuAGAAGuccAcuAccTsT942(781C) stab111484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1502L21 siNA antisenseAGGuAGAuGGuucGAGAAcTsT943(1484C) stab111538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1556L21 siNA antisenseuAucuccAucccuAAuAGGTsT944(1538C) stab11315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 333L21 siNA antisenseucAucGccuAcGuuGAAGGTsT945(315C) stab11355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 373L21 siNA antisenseAcGcGAGcAGAAcucAAAGTsT946(355C) stab11361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 361U21 siNA sense stab18B GuucuGcucGcGuuAcAcATT B947849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 849U21 siNA sense stab18B GAuuGGGcuGcuuuAAccuTT B948768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 768U21 siNA sense stab18B cGGuuAucAAccAGGuAGuTT B949781CAGGUAGUGGACUUCUACCAACC894HDAC3: 781U21 siNA sense stab18B GGuAGuGGAcuucuAccAATT B9501484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1484U21 siNA senseB GuucucGAAccAucuAccuTT B951stab181538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1538U21 siNA senseB ccuAuuAGGGAuGGAGAuATT B952stab18315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 315U21 siNA sense stab18B ccuucAAcGuAGGcGAuGATT B953355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 355U21 siNA sense stab18B cuuuGAGuucuGcucGcGuTT B954361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 379L21 siNA antisenseuGuGuAAcGcGAGcAGAAcTsT955(361C) stab08849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 867L21 siNA antisenseAGGuuAAAGcAGcccAAucTsT956(849C) stab08768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 786L21 siNA antisenseAcuAccuGGuuGAuAAccGTsT957(768C) stab08781CAGGUAGUGGACUUCUACCAACC894HDAC3: 799L21 siNA antisenseuuGGuAGAAGuccAcuAccTsT958(781C) stab081484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1502L21 siNA antisenseAGGuAGAuGGuucGAGAAcTsT959(1484C) stab081538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1556L21 siNA antisenseuAucuccAucccuAAuAGGTsT960(1538C) stab08315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 333L21 siNA antisenseucAucGccuAcGuuGAAGGTsT961(315C) stab08355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 373L21 siNA antisenseAcGcGAGcAGAAcucAAAGTsT962(355C) stab08361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 361U21 siNA sense stab09B GUUCUGCUCGCGUUACACATT B963849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 849U21 siNA sense stab09B GAUUGGGCUGCUUUAACCUTT B964768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 768U21 siNA sense stab09B CGGUUAUCAACCAGGUAGUTT B965781CAGGUAGUGGACUUCUACCAACC894HDAC3: 781U21 siNA sense stab09B GGUAGUGGACUUCUACCAATT B9661484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1484U21 siNA senseB GUUCUCGAACCAUCUACCUTT B967stab091538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1538U21 siNA senseB CCUAUUAGGGAUGGAGAUATT B968stab09315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 315U21 siNA sense stab09B CCUUCAACGUAGGCGAUGATT B969355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 355U21 siNA sense stab09B CUUUGAGUUCUGCUCGCGUTT B970361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 379L21 siNA antisenseUGUGUAACGCGAGCAGAACTsT971(361C) stab10849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 867L21 siNA antisenseAGGUUAAAGCAGCCCAAUCTsT972(849C) stab10768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 786L21 siNA antisenseACUACCUGGUUGAUAACCGTsT973(768C) stab10781CAGGUAGUGGACUUCUACCAACC894HDAC3: 799L21 siNA antisenseUUGGUAGAAGUCCACUACCTsT974(781C) stab101484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1502L21 siNA antisenseAGGUAGAUGGUUCGAGAACTsT975(1484C) stab101538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1556L21 siNA antisenseUAUCUCCAUCCCUAAUAGGTsT976(1538C) stab10315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 333L21 siNA antisenseUCAUCGCCUACGUUGAAGGTsT977(315C) stab10355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 373L21 siNA antisenseACGCGAGCAGAACUCAAAGTsT978(355C) stab10361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 379L21 siNA antisenseuGuGuAAcGcGAGcAGAAcTT B979(361C) stab19849UCGAUUGGGCUGCUUUAACCUCA892HDAC3:867L21 siNA antisenseAGGuuAAAGcAGcccAAucTT B980(849C) stab19768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 786L21 siNA antisenseAcuAccuGGuuGAuAAccGTT B981(768C) stab19781CAGGUAGUGGACUUCUACCAACC894HDAC3: 799L21 siNA antisenseuuGGuAGAAGuccAcuAccTT B982(781C) stab191484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1502L21 siNA antisenseAGGuAGAuGGuucGAGAAcTT B983(1484C) stab191538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1556L21 siNA antisenseuAucuccAucccuAAuAGGTT B984(1538C) stab19315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 333L21 siNA antisenseucAucGccuAcGuuGAAGGTT B985(315C) stab19355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 373L21 siNA antisenseAcGcGAGcAGAAcucAAAGTT B986(355C) stab19361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 379L21 siNA antisenseUGUGUAACGCGAGCAGAACTT B987(361C) stab22849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 867L21 siNA antisenseAGGUUAAAGCAGCCCAAUCTT B988(849C) stab22768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 786L21 siNA antisenseACUACCUGGUUGAUAACCGTT B989(768C) stab22781CAGGUAGUGGACUUCUACCAACC894HDAC3: 799L21 siNA antisenseUUGGUAGAAGUCCACUACCTT B990(781C) stab221484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1502L21 siNA antisenseAGGUAGAUGGUUCGAGAACTT B991(1484C) stab221538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1556L21 siNA antisenseUAUCUCCAUCCCUAAUAGGTT B992(1538C) stab22315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 333L21 siNA antisenseUCAUCGCCUACGUUGAAGGTT B993(315C) stab22355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 373L21 siNA antisenseACGCGAGCAGAACUCAAAGTT B994(355C) stab22361GAGUUCUGCUCGCGUUACACAGG891HDAC3: 379L21 siNA antisenseUGUGuAAcGcGAGcAGAAcTsT995(361C) stab25849UCGAUUGGGCUGCUUUAACCUCA892HDAC3: 867L21 siNA antisenseAGGuuAAAGcAGcccAAucTsT996(849C) stab25768GCCGGUUAUCAACCAGGUAGUGG893HDAC3: 786L21 siNA antisenseACUAccuGGuuGAuAAccGTsT997(768C) stab25781CAGGUAGUGGACUUCUACCAACC894HDAC3: 799L21 siNA antisenseUUGGuAGAAGuccAcuAccTsT998(781C) stab251484UGGUUCUCGAACCAUCUACCUGC895HDAC3: 1502L21 siNA antisenseAGGuAGAuGGuucGAGAAcTsT999(1484C) stab251538UACCUAUUAGGGAUGGAGAUACA896HDAC3: 1556L21 siNA antisenseUAUcuccAucccuAAuAGGTsT1000(1538C) stab25315UGCCUUCAACGUAGGCGAUGACU897HDAC3: 333L21 siNA antisenseUCAucGccuAcGuuGAAGGTsT1001(315C) stab25355CUCUUUGAGUUCUGCUCGCGUUA898HDAC3: 373L21 siNA antisenseACGcGAGcAGAAcucAAAGTsT1002(355C) stab25HDAC45108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5108U21 siNA senseGUUACGAUCGGAAUGCUUUTT19494373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4373U21 siNA senseGGCCGAGCUGCCGAAUUCATT19508280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8280U21 siNA senseGGUGAUGUAUGGCUAAGAUTT1951719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 719U21 siNA senseGCUCGUUGGAGCUAUCGUUTT19525829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5829U21 siNA senseGAGGGACCGUAGGUCUUUUTT1953720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 720U21 siNA senseCUCGUUGGAGCUAUCGUUUTT19547892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7892U21 siNA senseGUUUGCGUCUUAUUGAACUTT19558196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8196U21 siNA senseGACGGUUUAUUCUGAUUGATT19565108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5126L21 siNA antisenseAAAGCAUUGCGAUCGUAACTT1957(5108C)4373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4391L21 siNA antisenseUGAAUUCGGCAGCUCGGCCTT1958(4373C)8280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8298L21 siNA antisenseAUCUUAGCCAUACAUCACCTT1959(8280C)719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 737L21 siNA antisenseAACGAUAGCUCCAACGAGCTT1960(719C)5829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5847L21 siNA antisenseAAAAGACCUACGGUCCCUCTT1961(5829C)720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 738L21 siNA antisenseAAACGAUAGCUCCAACGAGTT1962(720C)7892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7910L21 siNA antisenseAGUUCAAUAAGACGCAAACTT1963(7892C)8196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8214L21 siNA antisenseUCAAUCAGAAUAAACCGUCTT1964(8196C)5108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5108U21 siNA senseB GuuAcGAucGGAAuGcuuuTT B1965stab044373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4373U21 siNA senseB GGccGAGcuGccGAAuucATT B1966stab048280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8280U21 siNA senseB GGuGAuGuAuGGcuAAGAuTT B1967stab04719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 719U21 siNA sense stab04B GcucGuuGGAGcuAucGuuTT B19685829AGGAGGGACGGUAGGUCUUUUCG1945HDAC4: 5829U21 siNA senseB GAGGGAccGuAGGucuuuuTT B1969stab04720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 720U21 siNA sense stab04B cucGuuGGAGcuAucGuuuTT B19707892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7892U21 siNA senseB GuuuGcGucuuAuuGAAcuTT B1971stab048196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8196U21 siNA senseB GAcGGuuuAuucuGAuuGATT B1972stab045108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5126L21 siNA antisenseAAAGcAuuccGAucGuAAcTsT1973(5108C) stab054373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4391L21 siNA antisenseuGAAuucGGcAGcucGGccTsT1974(4373C) stab058280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8298L21 siNA antisenseAucuuAGccAuAcAucAccTsT1975(8280C) stab05719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 737L21 siNA antisenseAAcGAuAGcuccAAcGAGcTsT1976(719C) stab055829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5847L21 siNA antisenseAAAAGAccuAcGGucccucTsT1977(5829C) stab05720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 738L21 siNA antisense(720C) stab05AAAcGAuAGcuccAAcGAGTsT19787892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7910L21 siNA antisenseAGuucAAuAAGAcGcAAAcTsT1979(7892C) stab058196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8214L21 siNA antisenseucAAucAGAAuAAAccGucTsT1980(8196C) stab055108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5108U21 siNA senseB GuuAcGAucGGAAuGcuuuTT B1981stab074373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4373U21 siNA senseB GGccGAGcuGccGAAuucATT B1982stab078280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8280U21 siNA senseB GGuGAuGuAuGGcuAAGAuTT B1983stab07719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 719U21 siNA sense stab07B GcucGuuGGAGcuAucGuuTT B19845829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5829U21 siNA senseB GAGGGAccGuAGGucuuuuTT B1985stab07720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 720U21 siNA sense stab07B cucGuuGGAGcuAucGuuuTT B19867892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7892U21 siNA senseB GuuuGcGucuuAuuGAAcuTT B1987stab078196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8196U21 siNA senseB GAcGGuuuAuucuGAuuGATT B1988stab075108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5126L21 siNA antisenseAAAGcAuuccGAucGuAAcTsT1989(5108C) stab114373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4391L21 siNA antisenseuGAAuucGGcAGcucGGccTsT1990(4373C) stab118280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8298L21 siNA antisenseAucuuAGccAuAcAucAccTsT1991(8280C) stab11719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 737L21 siNA antisenseAAcGAuAGcuccAAcGAGcTsT1992(719C) stab115829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5847L21 siNA antisenseAAAAGAccuAcGGucccucTsT1993(5829C) stab11720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 738L21 siNA antisenseAAAcGAuAGcuccAAcGAGTsT1994(720C) stab117892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7910L21 siNA antisenseAGuucAAuAAGAcGcAAAcTsT1995(7892C) stab118196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8214L21 siNA antisenseucAAucAGAAuAAAccGucTsT1996(8196C) stab115108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5108U21 siNA senseB GuuAcGAucGGAAuGcuuuTT B1997stab184373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4373U21 siNA senseB GGccGAGcuGccGAAuucATT B1998stab188280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8280U21 siNA senseB GGuGAuGuAuGGcuAAGAuTT B1999stab18719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 719U21 siNA sense stab18B GcucGuuGGAGcuAucGuuTT B20005829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5829U21 siNA senseB GAGGGAccGuAGGucuuuuTT B2001stab18720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 720U21 siNA sense stab18B cucGuuGGAGcuAucGuuuTT B20027892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7892U21 siNA senseB GuuuGcGucuuAuuGAAcuTT B2003stab188196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8196U21 siNA senseB GAcGGuuuAuucuGAuuGATT B2004stab185108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5126L21 siNA antisenseAAAGcAuuccGAucGuAAcTsT2005(5108C) stab084373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4391L21 siNA antisenseuGAAuucGGcAGcucGGccTsT2006(4373C) stab088280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8298L21 siNA antisenseAucuuAGccAuAcAucAccTsT2007(8280C) stab08719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 737L21 siNA antisenseAAcGAuAGcuccAAcGAGcTsT2008(719C) stab085829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5847L21 siNA antisenseAAAAGAccuAcGGucccucTsT2009(5829C) stab08720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 738L21 siNA antisenseAAAcGAuAGcuccAAcGAGTsT2010(720C) stab087892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7910L21 siNA antisenseAGuucAAuAAGAcGcAAAcTsT2011(7892C) stab088196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8214L21 siNA antisenseucAAucAGAAuAAAccGucTsT2012(8196C) stab085108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5108U21 siNA senseB GUUACGAUCGGAAUGCUUUTT B2013stab094373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4373U21 siNA senseB GGCCGAGCUGCCGAAUUCATT B2014stab098280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8280U21 siNA senseB GGUGAUGUAUGGCUAAGAUTT B2015stab09719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 719U21 siNA sense stab09B GCUCGUUGGAGCUAUCGUUTT B20165829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5829U21 siNA senseB GAGGGACCGUAGGUCUUUUTT B2017stab09720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 720U21 siNA sense stab09B CUCGUUGGAGCUAUCGUUUTT B20187892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7892U21 siNA senseB GUUUGCGUCUUAUUGAACUTT B2019stab098196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8196U21 siNA senseB GACGGUUUAUUCUGAUUGATT B2020stab095108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5126L21 siNA antisenseAAAGCAUUCCGAUCGUAACTsT2021(5108C) stab104373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4391L21 siNA antisenseUGAAUUCGGCAGCUCGGCCTsT2022(4373C) stab108280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8298L21 siNA antisenseAUCUUAGCCAUACAUCACCTsT2023(8280C) stab10719GAGCUCGUUGGAG0UAUCGUUUC1944HDAC4: 737L21 siNA antisenseAACGAUAGCUCCAACGAGCTsT2024(719C) stab105829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5847L21 siNA antisenseAAAAGACCUACGGUCCCUCTsT2025(5829C) stab10720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 738L21 siNA antisenseAAACGAUAGCUCCAACGAGTsT2026(720C) stab107892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7910L21 siNA antisenseAGUUCAAUAAGACGCAAACTsT2027(7892C) stab108196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8214L21 siNA antisenseUCAAUCAGAAUAAACCGUCTsT2028(8196C) stab105108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5126L21 siNA antisenseAAAGcAuuccGAucGuAAcTT B2029(5108C) stab194373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4391L21 siNA antisenseuGAAuucGGcAGcucGGccTT B2030(4373C) stab198280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8298L21 siNA antisenseAucuuAGccAuAcAucAccTT B2031(8280C) stab19719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 737L21 siNA antisenseAAcGAuAGcuccAAcGAGcTT B2032(719C) stab195829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5847L21 siNA antisenseAAAAGAccuAcGGucccucTT B2033(5829C) stab19720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 738L21 siNA antisenseAAAcGAuAGcuccAAcGAGTT B2034(720C) stab197892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7910L21 siNA antisenseAGuucAAuAAGAcGcAAAcTT B2035(7892C) stab198196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8214L21 siNA antisenseucAAucAGAAuAAAccGucTT B2036(8196C) stab195108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5126L21 siNA antisenseAAAGCAUUCCGAUCGUAACTT B2037(5108C) stab224373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4391L21 siNA antisenseUGAAUUCGGCAGCUCGGCCTT B2038(4373C) stab228280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8298L21 siNA antisenseAUCUUAGCCAUACAUCACCTT B2039(8280C) stab22719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 737L21 siNA antisenseAACGAUAGCUCCAACGAGCTT B2040(719C) stab225829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5847L21 siNA antisenseAAAAGACCUACGGUCCCUCTT B2041(5829C) stab22720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 738L21 siNA antisenseAAACGAUAGCUCCAACGAGTT B2042(720C) stab227892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7910L21 siNA antisenseAGUUCAAUAAGACGCAAACTT B2043(7892C) stab228196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8214L21 siNA antisenseUCAAUCAGAAUAAACCGUCTT B2044(8196C) stab225108GUGUUACGAUCGGAAUGCUUUUU1941HDAC4: 5126L21 siNA antisenseAAAGcAuuccGAucGuAAcTsT2045(5108C) stab254373GCGGCCGAGCUGCCGAAUUCAGU1942HDAC4: 4391L21 siNA antisenseUGAAuucGGcAGcucGGccTsT2046(4373C) stab258280UAGGUGAUGUAUGGCUAAGAUUU1943HDAC4: 8298L21 siNA antisenseAUCuuAGccAuAcAucAccTsT2047(8280C) stab25719GAGCUCGUUGGAGCUAUCGUUUC1944HDAC4: 737L21 siNA antisenseAACGAuAGcuccAAcGAGcTsT2048(719C) stab255829AGGAGGGACCGUAGGUCUUUUCG1945HDAC4: 5847L21 siNA antisenseAAAAGAccuAcGGucccucTsT2049(5829C) stab25720AGCUCGUUGGAGCUAUCGUUUCC1946HDAC4: 738L21 siNA antisenseAAAcGAuAGcuccAAcGAGTsT2050(720C) stab257892AAGUUUGCGUCUUAUUGAACUUA1947HDAC4: 7910L21 siNA antisenseAGUucAAuAAGAcGcAAAcTsT2051(7892C) stab258196GUGACGGUUUAUUCUGAUUGAGA1948HDAC4: 8214L21 siNA antisenseUCAAucAGAAuAAAccGucTsT2052(8196C) stab25HDAC51771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1771U21 siNA senseGCAUGCGGACGGUAGGCAATT26513771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3771U21 siNA senseGUCACACAUUCAACAAGGUTT2652321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 321U21 siNA senseCCCAGAGCCGGCAUGAACUTT26531031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1031U21 siNA senseGACGCCUCCCUCCUACAAATT26541182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1182U21 siNA senseCGCAAGGAUGGGACUGUUATT26551251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1251U21 siNA senseGCGUCGUCCGUGUGUAACATT26561567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1567U21 siNA senseCGCUGACCGGCAAGUUCAUTT26572196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2196U21 siNA senseCCUGGUGCUGGAUACAAAATT26581771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1789L21 siNA antisenseUUGCCUACCGUCCGCAUGCTT2659(1771C)3771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3789L21 siNA antisenseACCUUGUUGAAUGUGUGACTT2660(3771C)321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 339L21 siNA antisenseAGUUCAUGCCGGCUCUGGGTT2661(321C)1031GGGACGCCUCCCUCCUACAAAGU2646HDAC5v1: 1049L21 siNA antisenseUUUGUAGGAGGGAGGCGUCTT2662(1031C)1182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1200L21 siNA antisenseUAACAGUCCCAUCCUUGCGTT2663(1182C)1251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1269L21 siNA antisenseUGUUACACACGGACGACGCTT2664(1251C)1567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1585L21 siNA antisenseAUGAACUUGCCGGUCAGCGTT2665(1567C)2196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2214L21 siNA antisenseUUUUGUAUCCAGCACCAGGTT2666(2196C)1771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1771U21 siNA senseB GcAuGcGGAcGGuAGGcAATT B2667stab043771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3771U21 siNA senseB GucAcAcAuucAAcAAGGuTT B2668stab04321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 321U21 siNA senseB cccAGAGccGGcAuGAAcuTT B2669stab041031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1031U21 siNA senseB GAcGccucccuccuAcAAATT B2670stab041182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1182U21 siNA senseB cGcAAGGAuGGGAcuGuuATT B2671stab041251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1251U21 siNA senseB GcGucGuccGuGuGuAAcATT B2672stab041567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1567U21 siNA senseB cGcuGAccGGcAAGuucAuTT B2673stab042196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2196U21 siNA senseB ccuGGuGcuGGAuAcAAAATT B2674stab041771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1789L21 siNA antisenseuuGccuAccGuccGcAuGcTsT2675(1771C) stab053771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3789L21 siNA antisenseAccuuGuuGAAuGuGuGAcTsT2676(3771C) stab05321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 339L21 siNA antisenseAGuucAuGccGGcucuGGGTsT2677(321C) stab051031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1049L21 siNA antisenseuuuGuAGGAGGGAGGcGucTsT2678(1031C) stab051182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1200L21 siNA antisenseuAAcAGucccAuccuuGcGTsT2679(1182C) stab051251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1269L21 siNA antisenseuGuuAcAcAcGGAcGAcGcTsT2680(1251C) stab051567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1585L21 siNA antisenseAuGAAcuuGccGGucAGcGTsT2681(1567C) stab052196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2214L21 siNA antisenseuuuuGuAuccAGcAccAGGTsT2682(2196C) stab051771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1771U21 siNA senseB GcAuGcGGAcGGuAGGcAATT B2683stab073771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3771U21 siNA senseB GucAcAcAuucAAcAAGGUTT B2684stab07321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 321U21 siNA senseB cccAGAGccGGcAuGAAcuTT B2685stab071031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1031U21 siNA senseB GAcGCcucccuccuAcAAATT B2686stab071182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1182U21 siNA senseB cGcAAGGAuGGGAcuGuuATT B2687stab071251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1251U21 siNA senseB GcGucGuccGuGuGuAAcATT B2688stab071567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1567U21 siNA senseB cGcuGAccGGcAAGuucAuTT B2689stab072196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2196U21 siNA senseB ccuGGuGcuGGAuAcAAAATT B2690stab071771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1789L21 siNA antisenseuuGccuAccGuccGcAuGcTsT2691(1771C) stab113771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3789L21 siNA antisenseAccuuGuuGAAuGuGuGAcTsT2692(3771C) stab11321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 339L21 siNA antisenseAGuucAuGccGGcucuGGGTsT2693(321C) stab111031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1049L21 siNA antisenseuuuGuAGGAGGGAGGcGucTsT2694(1031C) stab111182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1200L21 siNA antisenseuAAcAGucccAuccuuGcGTsT2695(1182C) stab111251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1269L21 siNA antisenseuGuuAcAcAcGGAcGAcGcTsT2696(1251C) stab111567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1585L21 siNA antisenseAuGAAcuuGccGGucAGcGTsT2697(1567C) stab112196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2214L21 siNA antisenseuuuuGuAuccAGcAccAGGTsT2698(2196C) stab111771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1771U21 siNA senseB GcAuGcGGAcGGuAGGcAATT B2699stab183771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3771U21 siNA senseB GucAcAcAuucAAcAAGGuTT B2700stab18321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 321U21 siNA senseB cccAGAGccGGcAuGAAcuTT B2701stab181031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1031U21 siNA senseB GAcGccucccuccuAcAAATT B2702stab181182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1182U21 siNA senseB cGcAAGGAuGGGAcuGuuATT B2703stab181251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1251U21 siNA senseB GcGucGuccGuGuGuAAcATT B2704stab181567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1567U21 siNA senseB cGcuGAccGGcAAGuucAuTT B2705stab182196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2196U21 siNA senseB ccuGGuGcuGGAuAcAAAATT B2706stab181771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1789L21 siNA antisenseuuGccuAccGuccGcAuGcTsT2707(1771C) stab083771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3789L21 siNA antisenseAccuuGuuGAAuGuGuGAcTsT2708(3771C) stab08321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 339L21 siNA antisenseAGuucAuGccGGcucuGGGTsT2709(321C) stab081031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1049L21 siNA antisenseuuuGuAGGAGGGAGGcGucTsT2710(1031C) stab081182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1200L21 siNA antisenseuAAcAGucccAuccuuGcGTsT2711(1182C) stab081251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1269L21 siNA antisenseuGuuAcAcAcGGAcGAcGcTsT2712(1251C) stab081567CACGCUGACCGGCMGUUCAUGA2649HDAC5v1: 1585L21 siNA antisenseAuGAAcuuGccGGucAGcGTsT2713(1567C) stab082196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1:2214L21 siNA antisenseuuuuGuAuccAGcAccAGGTsT2714(2196C) stab081771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1771U21 siNA senseB GCAUGCGGACGGUAGGCAATT B2715stab093771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3771U21 siNA senseB GUCACACAUUCAACAAGGUTT B2716stab09321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 321U21 siNA senseB CCCAGAGCCGGCAUGAACUTT B2717stab091031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1031U21 siNA senseB GACGCCUCCCUCCUACAAATT B2718stab091182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1182U21 siNA senseB CGCAAGGAUGGGACUGUUATT B2719stab091251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1251U21 siNA senseB GCGUCGUCCGUGUGUAACATT B2720stab091567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1567U21 siNA senseB CGCUGACCGGCAAGUUCAUTT B2721stab092196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2196U21 siNA senseB CCUGGUGCUGGAUACAAAATT B2722stab091771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1789L21 siNA antisenseUUGCCUACCGUCCGCAUGCTsT2723(1771C) stab103771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3789L21 siNA antisenseACCUUGUUGAAUGUGUGACTsT2724(3771C) stab10321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 339L21 siNA antisenseAGUUCAUGCCGGCUCUGGGTsT2725(321C) stab101031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1049L21 siNA antisenseUUUGUAGGAGGGAGGCGUCTsT2726(1031C) stab101182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1200L21 siNA antisenseUAACAGUCCCAUCCUUGCGTsT2727(1182C) stab101251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1269L21 siNA antisenseUGUUACACACGGACGACGCTsT2728(1251C) stab101567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1585L21 siNA antisenseAUGAACUUGCCGGUCAGCGTsT2729(1567C) stab102196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2214L21 siNA antisenseUUUUGUAUCCAGCACCAGGTsT2730(2196C) stab101771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1789L21 siNA antisenseuuGccuAccGuccGcAuGcTT B2731(1771C) stab193771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3789L21 siNA antisenseAccuuGuuGAAuGuGuGAcTT B2732(3771C) stab19321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 339L21 siNA antisenseAGuucAuGccGGcucuGGGTT B2733(321C) stab191031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1049L21 siNA antisenseuuuGuAGGAGGGAGGcGucTT B2734(1031C) stab191182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1200L21 siNA antisenseuAAcAGucccAuccuuGcGTT B2735(1182C) stab191251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1269L21 siNA antisenseuGuuAcAcAcGGAcGAcGcTT B2736(1251C) stab191567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1585L21 siNA antisenseAuGAAcuuGccGGucAGcGTT B2737(1567C) stab192196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2214L21 siNA antisenseuuuuGuAuccAGcAccAGGTT B2738(2196C) stab191771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1789L21 siNA antisenseUUGCCUACCGUCCGCAUGCTT B2739(1771C) stab223771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3789L21 siNA antisenseACCUUGUUGAAUGUGUGACTT B2740(3771C) stab22321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 339L21 siNA antisenseAGUUCAUGCCGGCUCUGGGTT B2741(321C) stab221031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1049L21 siNA antisenseUUUGUAGGAGGGAGGCGUCTT B2742(1031C) stab221182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1200L21 siNA antisenseUAACAGUCCCAUCCUUGCGTT B2743(1182C) stab221251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1269L21 siNA antisenseUGUUACACACGGACGACGCTT B2744(1251C) stab221567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1585L21 siNA antisenseAUGAACUUGCCGGUCAGCGTT B2745(1567C) stab222196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2214L21 siNA antisenseUUUUGUAUCCAGCACCAGGTT B2746(2196C) stab221771CAGCAUGCGGACGGUAGGCAAGC2643HDAC5v1: 1789L21 siNA antisenseUUGccuAccGuccGcAuGcTsT2747(1771C) stab253771AAGUCACACAUUCAACAAGGUGU2644HDAC5v1: 3789L21 siNA antisenseACCuuGuuGAAuGuGuGAcTsT2748(3771C) stab25321GGCCCAGAGCCGGCAUGAACUCU2645HDAC5v1: 339L21 siNA antisenseAGUucAuGccGGcucuGGGTsT2749(321C) stab251031GGGACGCCUCCCUCCUACAAACU2646HDAC5v1: 1049L21 siNA antisenseUUUGuAGGAGGGAGGcGucTsT2750(1031C) stab251182GUCGCAAGGAUGGGACUGUUAUU2647HDAC5v1: 1200L21 siNA antisenseUAAcAGucccAuccuuGcGTsT2751(1182C) stab251251GGGCGUCGUCCGUGUGUAACAGC2648HDAC5v1: 1269L21 siNA antisenseUGUuAcAcAcGGAcGAcGcTsT2752(1251C) stab251567CACGCUGACCGGCAAGUUCAUGA2649HDAC5v1: 1585L21 siNA antisenseAUGAAcuuGccGGucAGcGTsT2753(1567C) stab252196AGCCUGGUGCUGGAUACAAAAAA2650HDAC5v1: 2214L21 siNA antisenseUUUuGuAuccAGcAccAGGTsT2754(2196C) stab25HDAC6825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 825U21 siNA senseCCGGAGGGUCCUUAUCGUATT32173904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3904U21 siNA senseGAGAACUGCGACGAUUAAUTT3218178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 178U21 siNA senseGUCACUUCGAAGCGAAAUATT32191540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1540U21 siNA senseCUGGUCUAUGACCAAAAUATT32201773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1773U21 siNA senseCCGUGAGAGUUCCAACUUUTT3221923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 923U21 siNA senseGCUACGAGCAGGGUAGGUUTT3222596AGCAGACACCUACGA0UCAGUUU3215HDAC6: 596U21 siNA senseCAGACACCUACGACUCAGUTT32232688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2688U21 siNA senseGCGGAUGACCACACGAGAATT3224825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 843L21 siNA antisenseUACGAUAAGGACCCUCCGGTT3225(825C)3904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3922L21 siNA antisenseAUUAAUCGUCGCAGUUCUCTT3226(3904C) 178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 196L21 siNA antisenseUAUUUCGCUUCGAAGUGACTT3227(178C)1540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1558L21 siNA antisenseUAUUUUGGUCAUAGACCAGTT3228(1540C)1773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1791L21 siNA antisenseAAAGUUGGAACUCUCACGGTT3229(1773C)923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 941L21 siNA antisenseAACCUACCCUGCUCGUAGCTT3230(923C)596AGCAGACACCUACGACUCAGUUU3215HDAC6: 614L21 siNA antisenseACUGAGUCGUAGGUGUCUGTT3231(596C)2688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2706L21 siNA antisenseUUCUCGUGUGGUCAUCCGCTT3232(2688C)825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 825U21 siNA sense stab04B ccGGAGGGuccuuAucGuATT B32333904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3904U21 siNA senseB GAGAAcuGcGAcGAuuAAuTT B3234stab04178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 178U21 siNA sense stab04B GucAcuucGAAGcGAAAuATT B32351540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1540U21 siNA senseB cuGGucuAuGAccAAAAuATT B3236stab041773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1773U21 siNA senseB CCGuGAGAGuuccAAcuuuTT B3237stab04923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 923U21 siNA sense stab04B GcuAcGAGcAGGGuAGGuuTT B3238596AGCAGACACCUACGACUCAGUUU3215HDAC6: 596U21 siNA sense stab04B cAGAcAccuAcGAcucAGuTT B32392688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2688U21 siNA senseB GcGGAuGAccAcAcGAGAATT B3240stab04825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 843L21 siNA antisenseuAcGAuAAGGAcccuccGGTsT3241(825C) stab053904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3922L21 siNA antisenseAuuAAucGucGcAGuucucTsT3242(3904C) stab05178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 196L21 siNA antisenseuAuuucGcuucGAAGuGAcTsT3243(178C) stab051540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1558L21 siNA antisenseuAuuuuGGucAuAGAccAGTsT3244(1540C) stab051773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1791L21 siNA antisenseAAAGuuGGAAcucucAcGGTsT3245(1773C) stab05923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 941L21 siNA antisenseAAccuAcccuGcucGuAGcTsT3246(923C) stab05596AGCAGACACCUACGACUCAGUUU3215HDAC6: 614L21 siNA antisense(596C) stab05AcuGAGucGuAGGuGucuGTsT32472688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2706L21 siNA antisenseuucucGuGuGGucAuccGcTsT3248(2688C) stab05825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 825U21 siNA sense stab07B ccGGAGGGuccuuAucGuATT B32493904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3904U21 siNA senseB GAGAACuGCGACGAUUAAUTT B3250stab07178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 178U21 siNA sense stab07B GucAcuucGAAGcGAAAuATT B32511540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1540U21 siNA senseB cuGGucuAuGAccAAAAuATT B3252stab071773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1773U21 siNA senseB ccGuGAGAGuuccAAcuuuTT B3253stab07923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 923U21 siNA sense stab07B GcuAcGAGcAGGGuAGGuuTT B3254596AGCAGACACCUACGACUCAGUUU3215HDAC6: 596U21 siNA sense stab07B cAGAcAccuAcGAcucAGuTT B32552688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2688U21 siNA senseB GcGGAuGAccAcAcGAGAATT B3256stab07825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 843L21 siNA antisenseuAcGAuAAGGAcccuccGGTsT3257(825C) stab113904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3922L21 siNA antisenseAuuAAucGucGcAGuucucTsT3258(3904C) stab11178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 196L21 siNA antisenseuAuuucGcuucGAAGuGAcTsT3259(178C) stab111540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1558L21 siNA antisenseuAuuuuGGucAuAGAccAGTsT3260(1540C) stab111773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1791L21 siNA antisenseAAAGuuGGAAcucucAcGGTsT3261(1773C) stab11923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 941L21 siNA antisenseAAccuAcccuGcucGuAGcTsT3262(923C) stab11596AGCAGACACCUACGACUCAGUUU3215HDAC6: 614L21 siNA antisenseAcuGAGucGuAGGuGucuGTsT3263(596C) stab112688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2706L21 siNA antisenseuucucGuGuGGucAuccGcTsT3264(2688C) stab11825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 825U21 siNA sense stab18B ccGGAGGGuccuuAucGuATT B32653904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3904U21 siNA senseB GAGAAcuGcGAcGAuuAAuTT B3266stab18178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 178U21 siNA sense stab18B GucAcuucGAAGcGAAAuATT B32671540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1540U21 siNA senseB cuGGucuAuGAccAAAAuATT B3268stab181773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1773U21 siNA senseB ccGuGAGAGuuccAAcuuuTT B3269stab18923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 923U21 siNA sense stab18B GcuAcGAGcAGGGuAGGuuTT B3270596AGCAGACACCUACGACUCAGUUU3215HDAC6: 596U21 siNA sense stab18B cAGAcAccuAcGAcucAGuTT B32712688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2688U21 siNA senseB GcGGAuGAccAcAcGAGAATT B3272stab18825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 843L21 siNA antisenseuAcGAuAAGGAcccuccGGTsT3273(825C) stab083904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3922L21 siNA antisenseAuuAAucGucGcAGuucucTsT3274(3904C) stab08178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 196L21 siNA antisenseuAuuucGcuucGAAGuGAcTsT3275(178C) stab081540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1558L21 siNA antisenseuAuuuuGGucAuAGAccAGTsT3276(1540C) stab081773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1791L21 siNA antisenseAAAGuuGGAAcucucAcGGTsT3277(1773C) stab08923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 941L21 siNA antisenseAAccuAcccuGcucGuAGcTsT3278(923C) stab08596AGCAGACACCUACGACUCAGUUU3215HDAC6: 614L21 siNA antisenseAcuGAGucGuAGGuGucuGTsT3279(596C) stab082688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2706L21 siNA antisenseuucucGuGuGGucAuccGcTsT3280(2688C) stab08825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 825U21 siNA sense stab09B CCGGAGGGUCCUUAUCGUATT B32813904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3904U21 siNA senseB GAGAACUGCGACGAUUAAUTT B3282stab09178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 178U21 siNA sense stab09B GUCACUUCGAAGCGAAAUATT B32831540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1540U21 siNA senseB CUGGUCUAUGACCAAAAUATT B3284stab091773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1773U21 siNA senseB CCGUGAGAGUUCCAACUUUTT B3285stab09923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 923U21 siNA sense stab09B GCUACGAGCAGGGUAGGUUTT B3286596AGCAGACACCUACGACUCAGUUU3215HDAC6: 596U21 siNA sense stab09B CAGACACCUACGACUCAGUTT B32872688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2688U21 siNA senseB GCGGAUGACCACACGAGAATT B3288stab09825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 843L21 siNA antisenseUACGAUAAGGACCCUCCGGTsT3289(825C) stab103904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3922L21 siNA antisenseAUUAAUCGUCGCAGUUCUCTsT3290(3904C) stab10178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 196L21 sNA antisenseUAUUUCGCUUCGAAGUGACTsT3291(178C) stab101540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1558L21 siNA antisenseUAUUUUGGUCAUAGACCAGTsT3292(1540C) stab101773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1791L21 siNA antisenseAAAGUUGGAACUCUCACGGTsT3293(1773C) stab10923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 941L21 siNA antisenseAACCUACCCUGCUCGUAGCTsT3294(923C) stab10596AGCAGACACCUACGACUCAGUUU3215HDAC6: 614L21 siNA antisenseACUGAGUCGUAGGUGUCUGTsT3295(596C) stab102688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2706L21 siNA antisenseUUCUCGUGUGGUCAUCCGCTsT3296(2688C) stab10825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 843L21 siNA antisenseuAcGAuAAGGAcccuccGGTT B3297(825C) stab193904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3922L21 siNA antisenseAuuAAucGucGcAGuucucTT B3298(3904C) stab19178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 196L21 siNA antisenseuAuuucGcuucGAAGuGAcTT B3299(178C) stab191540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1558L21 siNA antisenseuAuuuuGGucAuAGAccAGTT B3300(1540C) stab191773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1791L21 siNA antisenseAAAGuuGGAAcucucAcGGTT B3301(1773C) stab19923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 941L21 siNA antisenseAAccuAcccuGcucGuAGcTT B3302(923C) stab19596AGCAGACACCUACGACUCAGUUU3215HDAC6: 614L21 siNA antisenseAcuGAGucGuAGGuGucuGTT B3303(596C) stab192688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2706L21 siNA antisenseuucucGuGuGGucAuccGcTT B3304(2688C) stab19825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 843L21 siNA antisenseUACGAUAAGGACCCUCCGGTT B3305(825C) stab223904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3922L21 siNA antisenseAUUAAUCGUCGCAGUUCUCTT B3306(3904C) stab22178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 196L21 siNA antisenseUAUUUCGCUUCGAAGUGACTT B3307(178C) stab221540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1558L21 siNA antisenseUAUUUUGGUCAUAGACCAGTT B3308(1540C) stab221773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1791L21 siNA antisenseAAAGUUGGAACUCUCACGGTT B3309(1773C) stab22923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 941L21 siNA antisenseAACCUACCCUGCUCGUAGCTT B3310(923C) stab22596AGCAGACACCUACGACUCAGUUU3215HDAC6: 614L21 siNA antisenseACUGAGUCGUAGGUGUCUGTT B3311(596C) stab222688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2706L21 siNA antisenseUUCUCGUGUGGUCAUCCGCTT B3312(2688C) stab22825AUCCGGAGGGUCCUUAUCGUAGA3209HDAC6: 843L21 siNA antisenseUACGAuAAGGAcccuccGGTsT3313(825C) stab253904AAGAGAACUGCGACGAUUAAUUG3210HDAC6: 3922L21 siNA antisenseAUUAAucGucGcAGuucucTsT3314(3904C) stab25178GUGUCACUUCGAAGCGAAAUAUU3211HDAC6: 196L21 siNA antisenseUAUuucGcuucGAAGuGAcTsT3315(178C) stab251540GGCUGGUCUAUGACCAAAAUAUG3212HDAC6: 1558L21 siNA antisenseUAUuuuGGucAuAGAccAGTsT3316(1540C) stab251773CACCGUGAGAGUUCCAACUUUGA3213HDAC6: 1791L21 siNA antisenseAAAGuuGGAAcucucAcGGTsT3317(1773C) stab25923CCGCUACGAGCAGGGUAGGUUCU3214HDAC6: 941L21 siNA antisenseAACcuAcccuGcucGuAGcTsT3318(923C) stab25596AGCAGACACCUACGACUCAGUUU3215HDAC6: 614L21 siNA antisenseACUGAGucGuAGGuGucuGTsT3319(596C) stab252688GAGCGGAUGACCACACGAGAAAA3216HDAC6: 2706L21 siNA antisenseUUCucGuGuGGucAuccGcTsT3320(2688C) stab25HDAC716UGCAGGACCACGACAGGAUUAAG3667HDAC7: 16U21 siNA senseCAGGACCACGACAGGAUUATT367521GACCACGACAGGAUUAAGUGAGG3668HDAC7: 21U21 siNA senseCCACGACAGGAUUAAGUGATT3676370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 370U21 siNA senseCGCCGAUGCCCGAGUUGCATT3677476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 476U21 siNA senseGAAGCUAGCGGAGGUGAUUTT3678511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 511U21 siNA senseCGGCCCUAGAAAGAACAGUTT3679506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 506U21 siNA senseGCAGGCGGCCCUAGAAAGATT3680699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 699U21 siNA senseCGCUAUAAGCCCAAGAAGUTT36811243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1243U21 siNA senseCUCACGUCCAGGUGAUCAATT368216UGCAGGACCACGACAGGAUUAAG3667HDAC7: 34L21 siNA antisenseUAAUCCUGUCGUGGUCCUGTT3683(16C)21GACCACGACAGGAUUAAGUGAGG3668HDAC7: 39L21 siNA antisenseUCACUUAAUCCUGUCGUGGTT3684(21C)370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 388L21 siNA antisenseUGCAACUCGGGCAUCGGCGTT3685(370C)476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 494L21 siNA antisenseAAUCACCUCCGCUAGCUUCTT3686(476C)511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 529L21 siNA antisenseACUGUUCUUUCUAGGGCCGTT3687(511C)506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 524L21 siNA antisenseUGUUUCUAGGGCCGCCUGCTT3688(506C)699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 717L21 siNA antisenseACUUCUUGGGCUUAUAGCGTT3689(699C)1243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1261L21 siNA antisenseUUGAUCACCUGGACGUGAGTT3690(1243C)16UGCAGGACCACGACAGGAUUAAG3667HDAC7: 16U21 siNA sense stab04B cAGGAccAcGAcAGGAuuATT B369121GACCACGACAGGAUUAAGUGAGG3668HDAC7: 21U21 siNA sense stab04B ccAcGAcAGGAuuAAGuGATT B3692370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 370U21 siNA sense stab04B cGccGAuGcccGAGuuGcATT B3693476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 476U21 siNA sense stab04B GAAGcuAGcGGAGGuGAuuTT B3694511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 511U21 siNA sense stab04B cGGcccuAGAAAGAAcAGuTT B3695506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 506U21 siNA sense stab04B GcAGGcGGcccuAGAAAGATT B3696699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 699U21 siNA sense stab04B cGcuAuAAGcccAAGAAGuTT B36971243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1243U21 siNA senseB cucAcGuccAGGuGAucAATT B3698stab0416UGCAGGACCACGACAGGAUUAAG3667HDAC7: 34L21 siNA antisenseuAAuccuGucGuGGuccuGTsT3699(16C) stab0521GACCACGACAGGAUUAAGUGAGG3668HDAC7: 39L21 siNA antisenseucAcuuAAuccuGucGuGGTsT3700(21C) stab05370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 388L21 siNA antisenseuGcAAcucGGGcAucGGcGTsT3701(370C) stab05476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 494L21 siNA antisenseAAucAccuccGcuAGcuucTsT3702(476C) stab05511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 529L21 siNA antisenseAcuGuucuuucuAGGGccGTsT3703(511C) stab05506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 524L21 siNA antisenseucuuucuAGGGccGccuGcTsT3704(506C) stab05699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 717L21 siNA antisenseAcuucuuGGGcuuAuAGcGTsT3705(699C) stab051243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1261L21 siNA antisenseuuGAucAccuGGAcGuGAGTsT3706(1243C) stab0516UGCAGGACCACGACAGGAUUAAG3667HDAC7: 16U21 siNA sense stab07B cAGGAccAcGAcAGGAuuATT B370721GACCACGACAGGAUUAAGUGAGG3668HDAC7: 21U21 siNA sense stab07B ccAcGAcAGGAuuAAGuGATT B3708370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 370U21 siNA sense stab07B cGccGAuGcccGAGuuGcATT B3709476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 476U21 siNA sense stab07B GAAGcuAGcGGAGGuGAuuTT B3710511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 511U21 siNA sense stab07B cGGcccuAGAAAGAAcAGuTT B3711506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 506U21 siNA sense stab07B GcAGGcGGcccuAGAAAGATT B3712699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 699U21 siNA sense stab07B cGcuAuAAGcccAAGAAGuTT B37131243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1243U21 siNA senseB cucAcGuccAGGuGAucAATT B3714stab0716UGCAGGACCACGACAGGAUUAAG3667HDAC7: 34L21 siNA antisenseuAAuccuGucGuGGuccuGTsT3715(16C) stab1121GACCACGACAGGAUUAAGUGAGG3668HDAC7: 39L21 siNA antisenseucAcuuAAuccuGucGuGGTsT3716(21C) stab11370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 388L21 siNA antisenseuGcAAcucGGGcAucGGcGTsT3717(370C) stab11476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 494L21 siNA antisenseAAucAccuccGcuAGcuucTsT3718(476C) stab11511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 529L21 siNA antisenseAcuGuucuuucuAGGGccGTsT3719(511C) stab11506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 524L21 siNA antisenseucuuucuAGGGccGccuGcTsT3720(506C) stab11699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 717L21 siNA antisenseAcuucuuGGGcuuAuAGcGTsT3721(699C) stab111243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1261L21 siNA antisenseuuGAucAccuGGAcGuGAGTsT3722(1243C) stab1116UGCAGGACCACGACAGGAUUAAG3667HDAC7: 16U21 siNA sense stab18B cAGGAccAcGAcAGGAuuATT B372321GACCACGACAGGAUUAAGUGAGG3668HDAC7: 21U21 siNA sense stab18B ccAcGAcAGGAuuAAGuGATT B3724370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 370U21 siNA sense stab18B cGccGAuGcccGAGuuGcATT B3725476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 476U21 siNA sense stab18B GAAGcuAGcGGAGGuGAuuTT B3726511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 511U21 siNA sense stab18B cGGcccuAGAAAGAAcAGuTT B3727506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 506U21 siNA sense stab18B GcAGGcGGcccuAGAAAGATT B3728699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 699U21 siNA sense stab18B cGcuAuAAGcccAAGAAGuTT B37291243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1243U21 siNA senseB cucAcGuccAGGuGAucAATT B3730stab1816UGCAGGACCACGACAGGAUUAAG3667HDAC7: 34L21 siNA antisenseuAAuccuGucGuGGuccuGTsT3731(16C) stab0821GACCACGACAGGAUUAAGUGAGG3668HDAC7: 39L21 siNA antisenseucAcuuAAuccuGucGuGGTsT3732(21C) stab08370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 388L21 siNA antisenseuGcAAcucGGGcAucGGcGTsT3733(370C) stab08476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 494L21 siNA antisenseAAucAccuccGcuAGcuucTsT3734(476C) stab08511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 529L21 siNA antisenseAcuGuucuuucuAGGGccGTsT3735(511C) stab08506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 524L21 siNA antisenseucuuucuAGGGccGccuGcTsT3736(506C) stab08699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 717L21 siNA antisenseAcuucuuGGGcuuAuAGcGTsT3737(699C) stab081243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1261L21 siNA antisenseuuGAucAccuGGAcGuGAGTsT3738(1243C) stab0816UGCAGGACCACGACAGGAUUAAG3667HDAC7: 16U21 siNA sense stab09B CAGGACCACGACAGGAUUATT B373921GACCACGACAGGAUUAAGUGAGG3668HDAC7: 21U21 siNA sense stab09B CCACGACAGGAUUAAGUGATT B3740370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 370U21 siNA sense stab09B CGCCGAUGCCCGAGUUGCATT B3741476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 476U21 siNA sense stab09B GAAGCUAGCGGAGGUGAUUTT B3742511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 511U21 siNA sense stab09B CGGCCCUAGAAAGAACAGUTT B3743506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 506U21 siNA sense stab09B GCAGGCGGCCCUAGAAAGATT B3744699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 699U21 siNA sense stab09B CGCUAUAAGCCCAAGAAGUTT B37451243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1243U21 siNA senseB CUCACGUCCAGGUGAUCAATT B3746stab0916UGCAGGACCACGACAGGAUUAAG3667HDAC7: 34L21 siNA antisenseUAAUCCUGUCGUGGUCCUGTsT3747(16C) stab1021GACCACGACAGGAUUAAGUGAGG3668HDAC7: 39L21 siNA antisenseUCACUUAAUCCUGUCGUGGTsT3748(21C) stab10370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 388L21 siNA antisenseUGCAACUCGGGCAUGGGCGTsT3749(370C) stab 10476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 494L21 siNA antisenseAAUCACCUCCGCUAGCUUCTsT3750(476C) stab10511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 529L21 siNA antisenseACUGUUCUUUCUAGGGCCGTsT3751(511C) stab10506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 524L21 siNA antisenseUCUUUCUAGGGCCGCCUGCTsT3752(506C) stab10699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 717L21 siNA antisenseACUUCUUGGGCUUAUAGCGTsT3753(699C) stab101243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1261L21 siNA antisenseUUGAUCACCUGGACGUGAGTsT3754(1243C) stab1016UGCAGGACCACGACAGGAUUAAG3667HDAC7: 34L21 siNA antisenseuAAuccuGucGuGGuccuGTT B3755(16C) stab1921GACCACGACAGGAUUAAGUGAGG3668HDAC7: 39L21 siNA antisenseucAcuuAAuccuGucGuGGTT B3756(21C) stab19370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 388L21 siNA antisenseuGcAAcucGGGcAucGGcGTT B3757(370C) stab19476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 494L21 siNA antisenseAAucAccuccGcuAGcuucTT B3758(476C) stab19511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 529L21 siNA antisenseAcuGuucuuucuAGGGccGTT B3759(511C) stab19506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 524L21 siNA antisenseucuuucuAGGGccGccuGcTT B3760(506C) stab19699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 717L21 siNA antisenseAcuucuuGGGcuuAuAGcGTT B3761(699C) stab191243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1261L21 siNA antisenseuuGAucAccuGGAcGuGAGTT B3762(1243C) stab1916UGCAGGACCACGACAGGAUUAAG3667HDAC7: 34L21 siNA antisenseUAAUCCUGUCGUGGUCCUGTT B3763(16C) stab2221GACCACGACAGGAUUAAGUGAGG3668HDAC7: 39L21 siNA antisenseUCACUUAAUCCUGUCGUGGTT B3764(21C) stab22370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 388L21 siNA antisenseUGCAACUCGGGCAUCGGCGTT B3765(370C) stab22476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 494L21 siNA antisenseAAUCACCUCCGCUAGCUUCTT B3766(476C) stab22511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 529L21 siNA antisenseACUGUUCUUUCUAGGGCCGTT B3767(511C) stab22506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 524L21 siNA antisenseUCUUUCUAGGGCCGCCUGCTT B3768(506C) stab22699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 717L21 siNA antisenseACUUCUUGGGGUUAUAGCGTT B3769(699C) stab221243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1261L21 siNA antisenseUUGAUCACCUGGACGUGAGTT B3770(1243C) stab2216UGCAGGACCACGACAGGAUUAAG3667HDAC7: 34L21 siNA antisenseUAAuccuGucGuGGuccuGTsT3771(16C) stab2521GACCACGACAGGAUUAAGUGAGG3668HDAC7: 39L21 siNA antisenseUCAcuuAAuccuGucGuGGTsT3772(21C) stab25370CACGCCGAUGCCCGAGUUGCAGG3669HDAC7: 388L21 siNA antisenseUGCAAcucGGGcAucGGcGTsT3773(370C) stab25476CAGAAGCUAGCGGAGGUGAUUCU3670HDAC7: 494L21 siNA antisenseAAUcAccuccGcuAGcuucTsT3774(476C) stab25511GGCGGCCCUAGAAAGAACAGUCC3671HDAC7: 529L21 siNA antisenseACUGuucuuucuAGGGccGTsT3775(511C) stab25506CAGCAGGCGGCCCUAGAAAGAAC3672HDAC7: 524L21 siNAUCUuucuAGGGccGccuGcTsT3776(506C) stab25699UGCGCUAUAAGCCCAAGAAGUCC3673HDAC7: 717L21 siNA antisenseACUucuuGGGcuuAuAGcGTsT3777(699C) stab251243AACUCACGUCCAGGUGAUCAAGA3674HDAC7: 1261L21 siNA antisenseUUGAucAccuGGAcGuGAGTsT3778(1243C) stab25HDAC884CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 84U21 siNA senseGGUCCCGGUUUAUAUCUAUTT3979889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 889U21 siNA senseGGAAUUGGCAAGUGUCUUATT3980418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 418U21 siNA senseCUGAUUGACGGAAUGUGCATT3981426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 426U21 siNA senseCGGAAUGUGCAAAGUAGCATT3982923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 923U21 siNA senseGGCAGUUGGCAACACUCAUTT3983533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 533U21 siNA senseGAUUGCGACGGAAAUUUGATT3984542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 542U21 siNA senseGGAAAUUUGAGCGUAUUCUTT3985554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 554U21 siNA senseGUAUUCUCUACGUGGAUUUTT398684CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 102L21 siNA antisenseAUAGAUAUAAACCGGGACCTT3987(84C)889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 907L21 siNA antisenseUAAGACACUUGCCAAUUCCTT3988(889C)418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 436L21 siNA antisenseUGCACAUUCCGUCAAUCAGTT3989(418C)426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 444L21 siNA antisenseUGCUACUUUGCACAUUCCGTT3990(426C)923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 941L21 siNA antisenseAUGAGUGUUGCCAACUGCCTT3991(923C)533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 551L21 siNA antisenseUCAAAUUUCCGUCGCAAUCTT3992(533C)542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 560L21 siNA antisenseAGAAUACGCUCAAAUUUCCTT3993(542C)554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 572L21 siNA antisenseAAAUCCACGUAGAGAAUACTT3994(554C)84CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 84U21 siNA sense stab04B GGucccGGuuuAuAucuAuTT B3995889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 889U21 siNA sense stab04B GGAAuuGGcAAGuGucuuATT B3996418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 418U21 siNA sense stab04B cuGAuuGAcGGAAuGuGcATT B3997426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 426U21 siNA sense stab04B cGGAAuGuGcAAAGuAGcATT B3998923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 923U21 siNA sense stab04B GGcAGuuGGcAAcAcucAuTT B3999533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 533U21 siNA sense stab04B GAuuGcGAcGGAAAuuuGATT B4000542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 542U21 siNA sense stab04B GGAAAuuuGAGcGuAuucuTT B4001554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 554U21 siNA sense stab04B GuAuucucuAcGuGGAuuuTT B400284CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 102L21 siNA antisenseAuAGAuAuAAAccGGGAccTsT4003(84C) stab05889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 907L21 siNA antisenseuAAGAcAcuuGccAAuuccTsT4004(889C) stab05418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 436L21 siNA antisenseuGcAcAuuccGucAAucAGTsT4005(418C) stab05426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 444L21 siNA antisenseuGcuAcuuuGcAcAuuccGTsT4006(426C) stab05923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 941L21 siNA antisenseAuGAGuGuuGccAAcuGccTsT4007(923C) stab05533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 551L21 siNA antisenseucAAAuuuccGucGcAAucTsT4008(533C) stab05542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 560L21 siNA antisenseAGAAuAcGcucAAAuuuccTsT4009(542C) stab05554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 572L21 siNA antisenseAAAuccAcGuAGAGAAuAcTsT4010(554C) stab0584CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 84U21 siNA sense stab07B GGucccGGuuuAuAucuAuTT B4011889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 889U21 siNA sense stab07B GGAAuuGGcAAGuGucuuATT B4012418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 418U21 siNA sense stab07B cuGAuuGAcGGAAuGuGcATT B4013426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 426U21 siNA sense stab07B cGGAAuGuGcAAAGuAGcATT B4014923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 923U21 siNA sense stab07B GGcAGuuGGcAAcAcucAuTT B4015533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 533U21 siNA sense stab07B GAuuGcGAcGGAAAuuuGATT B4016542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 542U21 siNA sense stab07B GGAAAuuuGAGcGuAuucuTT B4017554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 554U21 siNA sense stab07B GuAuucucuAcGuGGAuuuTT B401884CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 102L21 siNA antisenseAuAGAuAuAAAccGGGAccTsT4019(84C) stab11889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 907L21 siNA antisenseuAAGAcAcuuGccAAuuccTsT4020(889C) stab11418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 436L21 siNA antisenseuGcAcAuuccGucAAucAGTsT4021(418C) stab11426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 444L21 siNA antisenseuGcuAcuuuGcAcAuuccGTsT4022(426C) stab11923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 941L21 siNA antisenseAuGAGuGuuGccAAcuGccTsT4023(923C) stab11533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 551L21 siNA antisenseucAAAuuuccGucGcAAucTsT4024(533C) stab11542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 560L21 siNA antisenseAGAAuAcGcucAAAuuuccTsT4025(542C) stab11554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 572L21 siNA antisenseAAAuccAcGuAGAGAAuAcTsT4026(554C) stab1184CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 84U21 siNA sense stab18B GGucccGGuuuAuAucuAuTT B4027889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 889U21 siNA sense stab18B GGAAuuGGcAAGuGucuuATT B4028418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 418U21 siNA sense stab18B cuGAuuGAcGGAAuGuGcATT B4029426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 426U21 siNA sense stab18B cGGAAuGuGcAAAGuAGcATT B4030923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 923U21 siNA sense stab18B GGcAGuuGGcAAcAcucAuTT B4031533ACGAUUGCGACGGAAAUUUGAGC3978HDAC8: 533U21 siNA sense stab18B GAuuGcGAcGGAAAuuuGATT B4032542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 542U21 siNA sense stab18B GGAAAuuuGAGcGuAuucuTT B4033554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 554U21 siNA sense stab18B GuAuucucuAcGuGGAuuuTT B403484CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 102L21 siNA antisenseAuAGAuAuAAAccGGGAccTsT4035(84C) stab08889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 907L21 siNA antisenseuAAGAcAcuuGccAAuuccTsT4036(889C) stab08418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 436L21 siNA antisenseuGcAcAuuccGucAAucAGTsT4037(418C) stab08426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 444L21 siNA antisenseuGcuAcuuuGcAcAuuccGTsT4038(426C) stab08923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 941L21 siNA antisenseAuGAGuGuuGccAAcuGccTsT4039(923C) stab08533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 551L21 siNA antisenseucAAAuuuccGucGcAAucTsT4040(533C) stab08542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 560L21 siNA antisenseAGAAuAcGcucAAAuuuccTsT4041(542C) stab08554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 572L21 siNA antisenseAAAuccAcGuAGAGAAuAcTsT4042(554C) stab0884CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 84U21 siNA sense stab09B GGUCCCGGUUUAUAUCUAUTT B4043889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 889U21 siNA sense stab09B GGAAUUGGCAAGUGUCUUATT B4044418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 418U21 siNA sense stab09B CUGAUUGACGGAAUGUGCATT B4045426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 426U21 siNA sense stab09B CGGAAUGUGCAAAGUAGCATT B4046923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 923U21 siNA sense stab09B GGCAGUUGGCAACACUCAUTT B4047533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 533U21 siNA sense stab09B GAUUGCGACGGAAAUUUGATT B4048542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 542U21 siNA sense stab09B GGAAAUUUGAGCGUAUUCUTT B4049554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 554U21 siNA sense stabo9B GUAUUCUCUACGUGGAUUUTT B405084CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 102L21 siNA antisenseAUAGAUAUAAACCGGGACCTsT4051(84C) stab10889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 907L21 siNA antisenseUAAGACACUUGCCAAUUCCTsT4052(889C) stab10418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 436L21 siNA antisenseUGCACAUUCCGUCAAUCAGTsT4053(418C) stab10426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 444L21 siNA antisenseUGCUACUUUGCACAUUCCGTsT4054(426C) stab10923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 941L21 siNA antisenseAUGAGUGUUGCCAACUGCCTsT4055(923C) stab10533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 551L21 siNA antisenseUCAAAUUUCCGUCGCAAUCTsT4056(533C) stab10542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 560L21 siNA antisenseAGAAUACGCUCAAAUUUCCTsT4057(542C) stab10554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 572L21 siNA antisenseAAAUCCACGUAGAGAAUACTsT4058(554C) stab1084CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 102L21 siNA antisenseAuAGAuAuAAAccGGGAccTT B4059(84C) stab19889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 907L21 siNA antisenseuAAGAcAcuuGccAAuuccTT B4060(889C) stab19418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 436L21 siNA antisenseuGcAcAuuccGucAAucAGTT B4061(418C) stab19426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 444L21 siNA antisenseuGcuAcuuuGcAcAuuccGTT B4062(426C) stab19923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 941L21 siNA antisenseAuGAGuGuuGccAAcuGccTT B4063(923C) stab19533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 551L21 siNA antisenseucAAAuuuccGucGcAAucTT B4064(533C) stab19542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 560L21 siNA antisenseAGAAuAcGcucAAAuuuccTT B4065(542C) stab19554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 572L21 siNA antisenseAAAuccAcGuAGAGAAuAcTT B4066(554C) stab1984CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 102L21 siNA antisenseAUAGAUAUAAACCGGGACCTT B4067(84C) stab22889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 907L21 siNA antisenseUAAGACACUUGCCAAUUCCTT B4068(889C) stab22418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 436L21 siNA antisenseUGCACAUUCCGUCAAUCAGTT B4069(418C) stab22426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 444L21 siNA antisenseUGCUACUUUGCACAUUCCGTT B4070(426C) stab22923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 941L21 siNA antisenseAUGAGUGUUGCCAACUGCCTT B4071(923C) stab22533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 551L21 siNA antisenseUCAAAUUUCCGUCGCAAUCTT B4072(533C) stab22542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 560L21 siNA antisenseAGAAUACGCUCAAAUUUCCTT B4073(542C) stab22554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 572L21 siNA antisenseAAAUCCACGUAGAGAAUACTT B4074(554C) stab2284CUGGUCCCGGUUUAUAUCUAUAG3971HDAC8: 102L21 siNA antisenseAUAGAuAuAAAccGGGAccTsT4075(84C) stab25889UGGGAAUUGGCAAGUGUCUUAAG3972HDAC8: 907L21 siNA antisenseUAAGAcAcuuGccAAuuccTsT4076(889C) stab25418GCCUGAUUGACGGAAUGUGCAAA3973HDAC8: 436L21 siNA antisenseUGCAcAuuccGucAAucAGTsT4077(418C) stab25426GACGGAAUGUGCAAAGUAGCAAU3974HDAC8: 444L21 siNA antisenseUGCuAcuuuGcAcAuuccGTsT4078(426C) stab25923AUGGCAGUUGGCAACACUCAUUU3975HDAC8: 941L21 siNA antisenseAUGAGuGuuGccAAcuGccTsT4079(923C) stab25533ACGAUUGCGACGGAAAUUUGAGC3976HDAC8: 551L21 siNA antisenseUCAAAuuuccGucGcAAucTsT4080(533C) stab25542ACGGAAAUUUGAGCGUAUUCUCU3977HDAC8: 560L21 siNA antisenseAGAAuAcGcucAAAuuuccTsT4081(542C) stab25554GCGUAUUCUCUACGUGGAUUUGG3978HDAC8: 572L21 siNA antisenseAAAuccAcGuAGAGAAuAcTsT4082(554C) stab25HDAC93108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3108U21 siNA senseCCAAAGCCCGAAUAUGAAUTT46073548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3548U21 siNA senseCGUAACCGCUGUGAUUCUATT46083573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3573U21 siNA senseCAGUAAACCACGAUUGGAATT46093822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3822U21 siNA senseGCUAUGAACGGAUCGUAAUTT4610987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 987U21 siNA senseCAAUGGGCCAACUGGAAGUTT46112292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2292U21 siNA senseCAGGAUACUCCUAGGUGAUTT46122294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2294U21 siNA senseGGAUACUCCUAGGUGAUGATT46133114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3114U21 siNA senseCCCGAAUAUGAAUGCUGUUTT46143108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3126L21 siNA antisenseAUUCAUAUUCGGGCUUUGGTT4615(3108C)3548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3566L21 siNA antisenseUAGAAUCACAGCGGUUACGTT4616(3548C)3573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3591L21 siNA antisenseUUCCAAUCGUGGUUUACUGTT4617(3573C)3822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3840L21 siNA antisenseAUUACGAUCGGUUCAUAGCTT4618(3822C)987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 1005L21 siNA antisenseACUUCCAGUUGGCCCAUUGTT4619(987C)2292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2310L21 siNA antisenseAUCACCUAGGAGUAUCCUGTT4620(2292C)2294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2312L21 siNA antisenseUCAUCACCUAGGAGUAUCCTT4621(2294C)3114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3132L21 siNA antisenseAACAGCAUUCAUAUUCGGGTT4622(3114C)3108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3108U21 siNA senseB ccAAAGcccGAAuAuGAAuTT B4623stab043548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3548U21 siNA senseB cGuAAccGcuGuGAuucuATT B4624stab043573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3573U21 siNA senseB cAGuAAAccAcGAuuGGAATT B4625stab043822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3822U21 siNA senseB GcuAuGAAcGGAucGuAAuTT B4626stab04987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 987U21 siNA senseB cAAuGGGccAAcuGGAAGuTT B4627stab042292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2292U21 siNA senseB cAGGAuAcuccuAGGuGAuTT B4628stab042294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2294U21 siNA senseB GGAuAcuccuAGGuGAuGATT B4629stab043114AGCCCGAAUAUGAAUGGUGUUAU4606HDAC9v4: 3114U21 siNA senseB cccGAAuAuGAAuGcuGuuTT B4630stab043108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3126L21 siNA antisenseAuucAuAuucGGGcuuuGGTsT4631(3108C) stab053548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3566L21 siNA antisenseuAGAAucAcAGcGGuuAcGTsT4632(3548C) stab053573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3591L21 siNA antisenseuuccAAucGuGGuuuAcuGTsT4633(3573C) stab053822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3840L21 siNA antisenseAuuAcGAuccGuucAuAGcTsT4634(3822C) stab05987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 1005L21 siNA antisenseAcuuccAGuuGGcccAuuGTsT4635(987C) stab052292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2310L21 siNA antisenseAucAccuAGGAGuAuccuGTsT4636(2292C) stab052294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2312L21 siNA antisenseucAucAccuAGGAGuAuccTsT4637(2294C) stab053114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3132L21 siNA antisenseAAcAGcAuucAuAuucGGGTsT4638(3114C) stab053108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3108U21 siNA senseB ccAAAGcccGAAuAuGAAuTT B4639stab073548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3548U21 siNA senseB cGuAAccGcuGuGAuucuATT B4640stab073573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3573U21 siNA senseB cAGuAAAccAcGAuuGGAATT B4641stab073822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3822U21 siNA senseB GcuAuGAAcGGAucGuAAuTT B4642stab07987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 987U21 siNA senseB cAAuGGGccAAcuGGAAGuTT B4643stab072292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2292U21 siNA senseB cAGGAuAcuccuAGGuGAuTT B4644stab072294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2294U21 siNA senseB GGAuAcuccuAGGuGAuGATT B4645stab073114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3114U21 siNA senseB cccGAAuAuGAAuGcuGuuTT B4646stab073108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3126L21 siNA antisenseAuucAuAuucGGGcuuuGGTsT4647(3108C) stab113548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3566L21 siNA antisenseuAGAAucAcAGcGGuuAcGTsT4648(3548C) stab113573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3591L21 siNA antisenseuuccAAucGuGGuuuAcuGTsT4649(3573C) stab113822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3840L21 siNA antisenseAuuAcGAuccGuucAuAGcTsT4650(3822C) stab11987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 1005L21 siNA antisenseAcuuccAGuuGGcccAuuGTsT4651(987C) stab112292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2310L21 siNA antisenseAucAccuAGGAGuAuccuGTsT4652(2292C) stab112294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2312L21 siNA antisenseucAucAccuAGGAGuAuccTsT4653(2294C) stab113114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3132L21 siNA antisenseAAcAGcAuucAuAuucGGGTsT4654(3114C) stab113108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3108U21 siNA senseB ccAAAGcccGAAuAuGAAuTT B4655stab183548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3548U21 siNA senseB cGuAAccGcuGuGAuucuATT B4656stab183573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3573U21 siNA senseB cAGuAAAccAcGAuuGGAATT B4657stab183822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3822U21 siNA senseB GcuAuGAAcGGAucGuAAuTT B4658stab18987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 987U21 siNA senseB cAAuGGGccAAcuGGAAGuTT B4659stab182292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2292U21 siNA senseB cAGGAuAcuccuAGGuGAuTT B4660stab182294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2294U21 siNA senseB GGAuAcuccuAGGuGAuGATT B4661stab183114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3114U21 siNA senseB cccGAAuAuGAAuGcuGuuTT B4662stab183108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3126L21 siNA antisenseAuucAuAuucGGGcuuuGGTsT4663(3108C) stab083548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3566L21 siNA antisenseuAGAAucAcAGcGGuuAcGTsT4664(3548C) stab083573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3591L21 siNA antisenseuuccAAucGuGGuuuAcuGTsT4665(3573C) stab083822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3840L21 siNA antisenseAuuAcGAuccGuucAuAGcTsT4666(3822C) stab08987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 1005L21 siNA antisenseAcuuccAGuuGGcccAuuGTsT4667(987C) stab082292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2310L21 siNA antisenseAucAccuAGGAGuAuccuGTsT4668(2292C) stab082294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2312L21 siNA antisenseucAucAccuAGGAGuAuccTsT4669(2294C) stab083114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3132L21 siNA antisenseAAcAGcAuucAuAuucGGGTsT4670(3114C) stab083108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3108U21 siNA senseB CCAAAGCCCGAAUAUGAAUTT B4671stab093548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3548U21 siNA senseB CGUAACCGCUGUGAUUCUATT B4672stab093573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3573U21 siNA senseB CAGUAAACCACGAUUGGAATT B4673stab093822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3822U21 siNA senseB GCUAUGAACGGAUCGUAAUTT B4674stab09987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 987U21 siNA senseB CAAUGGGCCAACUGGAAGUTT B4675stab092292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2292U21 siNA senseB CAGGAUACUCCUAGGUGAUTT B4676stab092294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2294U21 siNA senseB GGAUACUCCUAGGUGAUGATT B4677stab093114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3114U21 siNA senseB CCCGAAUAUGAAUGCUGUUTT B4678stab093108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3126L21 siNA antisenseAUUCAUAUUCGGGCUUUGGTsT4679(3108C) stab103548AACGUAAGCGCUGUGAUUCUAGA4600HDAC9v4: 3566L21 siNA antisenseUAGAAUCACAGCGGUUACGTsT4680(3548C) stab103573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3591L21 siNA antisenseUUCCAAUCGUGGUUUACUGTsT4681(3573C) stab103822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3840L21 siNA antisenseAUUACGAUCCGUUCAUAGCTsT4682(3822C) stab10987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 1005L21 siNA antisenseACUUCCAGUUGGCCCAUUGTsT4683(987C) stab102292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2310L21 siNA antisenseAUCACCUAGGAGUAUCCUGTsT4684(2292C) stab102294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2312L21 siNA antisenseUCAUCACCUAGGAGUAUCCTsT4685(2294C) stab103114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3132L21 siNA antisenseAACAGCAUUCAUAUUCGGGTsT4686(3114C) stab103108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3126L21 sNA antisenseAuucAuAuucGGGcuuuGGTT B4687(3108C) stab193548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3566L21 siNA antisenseuAGAAucAcAGcGGuuAcGTT B4688(3548C) stab193573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3591L21 siNA antisenseuuccAAucGuGGuuuAcuGTT B4689(3573C) stab193822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3840L21 siNA antisenseAuuAcGAuccGuucAuAGcTT B4690(3822C) stab19987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 1005L21 siNA antisenseAcuuccAGuuGGcccAuuGTT B4691(987C) stab192292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2310L21 siNA antisenseAucAccuAGGAGuAuccuGTT B4692(2292C) stab192294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2312L21 siNA antisenseucAucAccuAGGAGuAuccTT B4693(2294C) stab193114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3132L21 siNA antisenseAAcAGcAuucAuAuucGGGTT B4694(3114C) stab193108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3126L21 siNA antisenseAUUCAUAUUCGGGCUUUGGTT B4695(3108C) stab223548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3566L21 siNA antisenseUAGAAUCACAGCGGUUACGTT B4696(3548C) stab223573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3591L21 siNA antisenseUUCCAAUCGUGGUUUACUGTT B4697(3573C) stab223822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4: 3840L21 siNA antisenseAUUACGAUCCGUUCAUAGCTT B4698(3822C) stab22987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 1005L21 siNA antisenseACUUCCAGUUGGCCCAUUGTT B4699(987C) stab222292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2310L21 siNA antisenseAUCACCUAGGAGUAUCCUGTT B4700(2292C) stab222294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2312L21 siNA antisenseUCAUCACCUAGGAGUAUCCTT B4701(2294C) stab223114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3132L21 siNA antisenseAACAGCAUUCAUAUUCGGGTT B4702(3114C) stab223108CACCAAAGCCCGAAUAUGAAUGC4599HDAC9v4: 3126L21 siNA antisenseAUUcAuAuucGGGcuuuGGTsT4703(3108C) stab253548AACGUAACCGCUGUGAUUCUAGA4600HDAC9v4: 3566L21 siNA antisenseUAGAAucAcAGcGGuuAcGTsT4704(3548C) stab253573UACAGUAAACCACGAUUGGAAGA4601HDAC9v4: 3591L21 siNA antisenseUUCcAAucGuGGuuuAcuGTsT4705(3573C) stab253822UAGCUAUGAACGGAUCGUAAUUC4602HDAC9v4:3840L21 siNA antisenseAUUAcGAuccGuucAuAGcTsT4706(3822C) stab25987AACAAUGGGCCAACUGGAAGUGU4603HDAC9v4: 1005L21 siNA antisenseACUuccAGuuGGcccAuuGTsT4707(987C) stab252292CCCAGGAUACUCCUAGGUGAUGA4604HDAC9v4: 2310L21 siNA antisenseAUCAccuAGGAGuAuccuGTsT4708(2292C) stab252294CAGGAUACUCCUAGGUGAUGACU4605HDAC9v4: 2312L21 siNA antisenseUCAucAccuAGGAGuAuccTsT4709(2294C) stab253114AGCCCGAAUAUGAAUGCUGUUAU4606HDAC9v4: 3132L21 siNA antisenseAACAGcAuucAuAuucGGGTsT4710(3114C) stab25HDAC11500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 500U21 siNA senseCGCUCGCCAUCAAGUUUCUTT4913777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 777U21 siNA senseCGACGUGGUGGUAUACAAUTT4914899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 899U21 siNA senseGCCGGGUGCCCAUCCUUAUTT4915957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 957U21 siNA senseCAUUGCUGACUCCAUACUUTT49161333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1333U21 siNA senseCAGGCAGUUAACUGAGAAUTT491719GCCCCGGGAUGCUACACACAACC4910HDAC11: 19U21 siNA senseCCCGGGAUGCUACACACAATT491879UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 79U21 siNA senseGUGUACUCGCCGCGCUACATT4919491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 491U21 siNA senseCGGACAUCACGCUCGCCAUTT4920500CACGCUCGCCAUCPAGUUUCUGU4905HDAC11: 518L21 siNA antisenseAGAAACUUGAUGGCGAGCGTT4921(500C)777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 795L21 siNA antisenseAUUGUAUACCACCACGUCGTT4922(777C)899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 917L21 siNA antisenseAUAAGGAUGGGCACCCGGCTT4923(899C)957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 975L21 siNA antisenseAAGUAUGGAGUCAGCAAUGTT4924(957C)1333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1351L21 siNA antisenseAUUCUCAGUUAACUGCCUGTT4925(1333C)19GCCCCGGGAUGCUACACACAACC4910HDAC11: 37L21 siNA antisenseUUGUGUGUAGCAUCCCGGGTT4926(19C)79UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 97L21 siNA antisenseUGUAGCGCGGCGAGUACACTT4927(79C)491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 509L21 siNA antisenseAUGGCGAGCGUGAUGUCCGTT4928(491C)500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 500U21 siNA senseB cGcucGccAucAAGuuucuTT B4929stab04777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 777U21 siNA senseB cGAcGuGGuGGuAuAcAAuTT B4930stab04899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 899U21 siNA senseB GccGGGuGcccAuccuuAuTT B4931stab04957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 957U21 siNA senseB cAuuGcuGAcuccAuAcuuTT B4932stab041333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1333U21 siNA senseB cAGGcAGuuAAcuGAGAAuTT B4933stab0419GCCCCGGGAUGCUACACACAACC4910HDAC11: 19U21 siNA sense stab04B cccGGGAuGcuAcAcAcAATT B493479UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 79U21 siNA sense stab04B GuGuAcucGccGcGcuAcATT B4935491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 491U21 siNA senseB cGGAcAucAcGcucGccAuTT B4936stab04500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 518L21 siNA antisenseAGAAAcuuGAuGGcGAGcGTsT4937(500C) stab05777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 795L21 siNA antisenseAuuGuAuAccAccAcGucGTsT4938(777C) stab05899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 917L21 siNA antisenseAuAAGGAuGGGcAcccGGcTsT4939(899C) stab05957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 975L21 siNA antisenseAAGuAuGGAGucAGcAAuGTsT4940(957C) stab051333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1351L21 siNA antisenseAuucucAGuuAAcuGccuGTsT4941(1333C) stab0519GCCCCGGGAUGCUACACACAACC4910HDAC11: 37L21 siNA antisenseuuGuGuGuAGcAucccGGGTsT4942(19C) stab0579UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 97L21 siNA antisenseuGuAGcGcGGcGAGuAcAcTsT4943(79C) stab05491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 509L21 siNA antisenseAuGGcGAGcGuGAuGuccGTsT4944(491C) stab05500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 500U21 siNA senseB cGcucGccAucAAGuuucuTT B4945stab07777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 777U21 siNA senseB cGAcGuGGuGGuAuAcAAuTT B4946stab07899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 899U21 siNA senseB GccGGGuGcccAuccuuAuTT B4947stab07957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 957U21 siNA senseB cAuuGcuGAcuccAuAcuuTT B4948stab071333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1333U21 siNA senseB cAGGcAGuuAAcuGAGAAuTT B4949stab0719GCCCCGGGAUGCUACACACAACC4910HDAC11: 19U21 siNA senseB cccGGGAuGcuAcAcAcAATT B4950stab0779UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 79U21 siNA senseB GuGuAcucGccGcGcuAcATT B4951stab07491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 491U21 siNA senseB cGGAcAucAcGcucGccAuTT B4952stab07500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 518L21 siNA antisenseAGAAAcuuGAuGGcGAGcGTsT4953(500C) stab11777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 795L21 siNA antisenseAuuGuAuAccAccAcGucGTsT4954(777C) stab11899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 917L21 siNA antisenseAuAAGGAuGGGcAcccGGcTsT4955(899C) stab11957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 975L21 siNA antisenseAAGuAuGGAGucAGcAAuGTsT4956(957C) stab111333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1351L21 siNA antisenseAuucucAGuuAAcuGccuGTsT4957(1333C) stab1119GCCCCGGGAUGCUACACACAACC4910HDAC11: 37L21 siNA antisenseuuGuGuGuAGcAucccGGGTsT4958(19C) stab1179UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 97L21 siNA antisenseuGuAGcGcGGcGAGuAcAcTsT4959(79C) stab11491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 509L21 siNA antisenseAuGGcGAGcGuGAuGuccGTsT4960(491C) stab11500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 500U21 siNA senseB cGcucGccAucAAGuuucuTT B4961stab18777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 777U21 siNA senseB cGAcGuGGuGGuAuAcAAuTT B4962stab18899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 899U21 siNA senseB GccGGGuGcccAuccuuAuTT B4963stab18957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 957U21 siNA senseB cAuuGcuGAcuccAuAcuuTT B4964stab181333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1333U21 siNA senseB cAGGcAGuuAAcuGAGAAuTT B4965stab1819GCCCCGGGAUGCUACACACAACC4910HDAC11: 19U21 siNA senseB cccGGGAuGcuAcAcAcAATT B4966stab1879UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 79U21 siNA senseB GuGuAcucGccGcGcuAcATT B4967stab18491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 491U21 siNA senseB cGGAcAucAcGcucGccAuTT B4968stab18500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 518L21 siNA antisenseAGAAAcuuGAuGGcGAGcGTsT4969(500C) stab08777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 795L21 siNA antisenseAuuGuAuAccAccAcGucGTsT4970(777C) stab08899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 917L21 siNA antisenseAuAAGGAuGGGcAcccGGcTsT4971(899C) stab08957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 975L21 siNA antisenseAAGuAuGGAGucAGcAAuGTsT4972(957C) stab081333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1351L21 siNA antisenseAuucucAGuuAAcuGccuGTsT4973(1333C) stab0819GCCCCGGGAUGCUACACACAACC4910HDAC11: 37L21 siNA antisenseuuGuGuGuAGcAucccGGGTsT4974(19C) stab0879UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 97L21 siNA antisenseuGuAGcGcGGcGAGuAcAcTsT4975(79C) stab08491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 509L21 siNA antisenseAuGGcGAGcGuGAuGuccGTsT4976(491C) stab08500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 500U21 siNA senseB CGCUCGCCAUCAAGUUUCUTT B4977stab09777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 777U21 siNA senseB CGACGUGGUGGUAUACAAUTT B4978stab09899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 899U21 siNA senseB GCCGGGUGCCCAUCCUUAUTT B4979stab09957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 957U21 siNA senseB CAUUGCUGACUCCAUACUUTT B4980stab091333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1333U21 siNA senseB CAGGCAGUUAACUGAGAAUTT B4981stab0919GCCCCGGGAUGCUACACACAACC4910HDAC11: 19U21 siNA sense stab09B CCCGGGAUGCUACACACAATT B498279UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 79U21 siNA sense stab09B GUGUACUCGCCGCGCUACATT B4983491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 491U21 siNA senseB CGGACAUCACGCUCGCCAUTT B4984stab09500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 518L21 siNA antisenseAGAAACUUGAUGGCGAGCGTsT4985(500C) stab10777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 795L21 siNA antisenseAUUGUAUACCACCACGUCGTsT4986(777C) stab10899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 917L21 siNA antisenseAUAAGGAUGGGCACCCGGCTsT4987(899C) stab10957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 975L21 siNA antisenseAAGUAUGGAGUCAGCAAUGTsT4988(957C) stab101333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1351L21 siNA antisenseAUUCUCAGUUAACUGCCUGTsT4989(1333C) stab1019GCCCCGGGAUGCUACACACAACC4910HDAC11: 137L21 siNA antisenseUUGUGUGUAGCAUCCCGGGTsT4990(19C) stab1079UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 97L21 siNA antisenseUGUAGCGCGGCGAGUACACTsT4991(79C) stab10491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 509L21 siNA antisenseAUGGCGAGCGUGAUGUCCGTsT4992(491C) stab10500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 518L21 siNA antisenseAGAAAcuuGAuGGcGAGcGTT B4993(500C) stab19777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 795L21 siNA antisenseAuuGuAuAccAccAcGucGTT B4994(777C) stab19899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 917L21 siNA antisenseAuAAGGAuGGGcAcccGGcTT B4995(899C) stab19957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 975L21 siNA antisenseAAGuAuGGAGucAGcAAuGTT B4996(957C) stab191333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1351L21 siNA antisenseAuucucAGuuAAcuGccuGTT B4997(1333C) stab1919GCCCCGGGAUGCUACACACAACC4910HDAC11: 37L21 siNA antisenseuuGuGuGuAGcAucccGGGTT B4998(19C) stab1979UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 97L21 siNA antisenseuGuAGcGcGGcGAGuAcAcTT B4999(79C) stab19491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 509L21 siNA antisenseAuGGcGAGcGuGAuGuccGTT B5000(491C) stab19500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 518L21 siNA antisenseAGAAACUUGAUGGCGAGCGTT B5001(500C) stab22777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 795L21 siNA antisenseAUUGUAUACCACCACGUCGTT B5002(777C) stab22899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 917L21 siNA antisenseAUAAGGAUGGGCACCCGGCTT B5003(899C) stab22957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 975L21 siNA antisenseAAGUAUGGAGUCAGCAAUGTT B5004(957C) stab221333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1351L21 siNA antisenseAUUCUCAGUUAACUGCCUGTT B5005(1333C) stab2219GCCCCGGGAUGCUACACACAACC4910HDAC11: 37L21 siNA antisenseUUGUGUGUAGCAUCCCGGGTT B5006(19C) stab2279UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 97L21 siNA antisenseUGUAGCGCGGCGAGUACACTT B5007(79C) stab22491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 509L21 siNA antisenseAUGGCGAGCGUGAUGUCCGTT B5008(491C) stab22500CACGCUCGCCAUCAAGUUUCUGU4905HDAC11: 518L21 siNA antisenseAGAAAcuuGAuGGcGAGcGTsT5009(500C) stab25777CCCGACGUGGUGGUAUACAAUGC4906HDAC11: 795L21 siNA antisenseAUUGuAuAccAccAcGucGTsT5010(777C) stab25899CCGCCGGGUGCCCAUCCUUAUGG4907HDAC11: 917L21 siNA antisenseAUAAGGAuGGGcAcccGGcTsT5011(899C) stab25957AUCAUUGCUGACUCCAUACUUAA4908HDAC11: 975L21 siNA antisenseAAGuAuGGAGucAGcAAuGTsT5012(957C) stab251333GGCAGGCAGUUAACUGAGAAUUG4909HDAC11: 1351L21 siNA antisenseAUUcucAGuuAAcuGccuGTsT5013(1333C) stab2519GCCCCGGGAUGCUACACACAACC4910HDAC11: 37L21 siNA antisenseUUGuGuGuAGcAucccGGGTsT5014(19C) stab2579UCGUGUACUCGCCGCGCUACAAC4911HDAC11: 97L21 siNA antisenseUGUAGcGcGGcGAGuAcAcTsT5015(79C) stab25491UGCGGACAUCACGCUCGCCAUCA4912HDAC11: 509L21 siNA antisenseAUGGcGAGcGuGAuGuccGTsT5016(491C) stab25
Uppercase = ribonucleotide

u,c = 2′-deoxy-2′-fluoro U,C

T = thymidine

B = inverted deoxy abasic

s = phosphorothioate linkage

A = deoxy Adenosine

G = deoxy Guanosine

G = 2′-O-methyl Guanosine

A = 2′-O-methyl Adenosine

TABLE IV

Non-limiting examples of Stabilization Chemistries

for chemically modified siNA constructs

Chemistry
pyrimidine
Purine
cap
p = S
Strand

“Stab 00”
Ribo
Ribo
TT at 3′-

S/AS

ends

“Stab 1”
Ribo
Ribo
—
5 at 5′-end
S/AS

1 at 3′-end

“Stab 2”
Ribo
Ribo
—
All linkages
Usually AS

“Stab 3”
2′-fluoro
Ribo
—
4 at 5′-end
Usually S

4 at 3′-end

“Stab 4”
2′-fluoro
Ribo
5′ and 3′-
—
Usually S

ends

“Stab 5”
2′-fluoro
Ribo
—
1 at 3′-end
Usually AS

“Stab 6”
2′-O-Methyl
Ribo
5′ and 3′-
—
Usually S

ends

“Stab 7”
2′-fluoro
2′-deoxy
5′ and 3′-
—
Usually S

ends

“Stab 8”
2′-fluoro
2′-O-
—
1 at 3′-end
S/AS

Methyl

“Stab 9”
Ribo
Ribo
5′ and 3′-
—
Usually S

ends

“Stab 10”
Ribo
Ribo
—
1 at 3′-end
Usually AS

“Stab 11”
2′-fluoro
2′-deoxy
—
1 at 3′-end
Usually AS

“Stab 12”
2′-fluoro
LNA
5′ and 3′-

Usually S

ends

“Stab 13”
2′-fluoro
LNA

1 at 3′-end
Usually AS

“Stab 14”
2′-fluoro
2′-deoxy

2 at 5′-end
Usually AS

1 at 3′-end

“Stab 15”
2′-deoxy
2′-deoxy

2 at 5′-end
Usually AS

1 at 3′-end

“Stab 16”
Ribo
2′-O-
5′ and 3′-

Usually S

Methyl
ends

“Stab 17”
2′-O-Methyl
2′-O-
5′ and 3′-

Usually S

Methyl
ends

“Stab 18”
2′-fluoro
2′-O-
5′ and 3′-

Usually S

Methyl
ends

“Stab 19”
2′-fluoro
2′-O-
3′-end

S/AS

Methyl

“Stab 20”
2′-fluoro
2′-deoxy
3′-end

Usually AS

“Stab 21”
2′-fluoro
Ribo
3′-end

Usually AS

“Stab 22”
Ribo
Ribo
3′-end

Usually AS

“Stab 23”
2′-fluoro*
2′-deoxy*
5′ and 3′-

Usually S

ends

“Stab 24”
2′-fluoro*
2′-O-
—
1 at 3′-end
S/AS

Methyl*

“Stab 25”
2′-fluoro*
2′-O-
—
1 at 3′-end
S/AS

Methyl*

“Stab 26”
2′-fluoro*
2′-O-
—

S/AS

Methyl*

“Stab 27”
2′-fluoro*
2′-O-
3′-end

S/AS

Methyl*

“Stab 28”
2′-fluoro*
2′-O-
3′-end

S/AS

Methyl*

“Stab 29”
2′-fluoro*
2′-O-

1 at 3′-end
S/AS

Methyl*

“Stab 30”
2′-fluoro*
2′-O-

S/AS

Methyl*

“Stab 31”
2′-fluoro*
2′-O-
3′-end

S/AS

Methyl*

“Stab 32”
2′-fluoro
2′-O-

S/AS

Methyl

“Stab 33”
2′-fluoro
2′-deoxy*
5′ and 3′-
—
Usually S

ends

“Stab 34”
2′-fluoro
2′-O-
5′ and 3′-

Usually S

Methyl*
ends

“Stab 3F”
2′-OCF3
Ribo
—
4 at 5′-end
Usually S

4 at 3′-end

“Stab 4F”
2′-OCF3
Ribo
5′ and 3′-
—
Usually S

ends

“Stab 5F”
2′-OCF3
Ribo
—
1 at 3′-end
Usually AS

“Stab 7F”
2′-OCF3
2′-deoxy
5′ and 3′-
—
Usually S

ends

“Stab 8F”
2′-OCF3
2′-O-
—
1 at 3′-end
S/AS

Methyl

“Stab 11F”
2′-OCF3
2′-deoxy
—
1 at 3′-end
Usually AS

“Stab 12F”
2′-OCF3
LNA
5′ and 3′-

Usually S

ends

“Stab 13F”
2′-OCF3
LNA

1 at 3′-end
Usually AS

“Stab 14F”
2′-OCF3
2′-deoxy

2 at 5′-end
Usually AS

1 at 3′-end

“Stab 15F”
2′-OCF3
2′-deoxy

2 at 5′-end
Usually AS

1 at 3′-end

“Stab 18F”
2′-OCF3
2′-O-
5′ and 3′-

Usually S

Methyl
ends

“Stab 19F”
2′-OCF3
2′-O-
3′-end

S/AS

Methyl

“Stab 20F”
2′-OCF3
2′-deoxy
3′-end

Usually AS

“Stab 21F”
2′-OCF3
Ribo
3′-end

Usually AS

“Stab 23F”
2′-OCF3*
2′-deoxy*
5′ and 3′-

Usually S

ends

“Stab 24F”
2′-OCF3*
2′-O-
—
1 at 3′-end
S/AS

Methyl*

“Stab 25F”
2′-OCF3*
2′-O-
—
1 at 3′-end
S/AS

Methyl*

“Stab 26F”
2′-OCF3*
2′-O-
—

S/AS

Methyl*

“Stab 27F”
2′-OCF3*
2′-O-
3′-end

S/AS

Methyl*

“Stab 28F”
2′-OCF3*
2′-O-
3′-end

S/AS

Methyl*

“Stab 29F”
2′-OCF3*
2′-O-

1 at 3′-end
S/AS

Methyl*

“Stab 30F”
2′-OCF3*
2′-O-

S/AS

Methyl*

“Stab 31F”
2′-OCF3*
2′-O-
3′-end

S/AS

Methyl*

“Stab 32F”
2′-OCF3
2′-O-

S/AS

Methyl

“Stab 33F”
2′-OCF3
2′-deoxy*
5′ and 3′-
—
Usually S

ends

“Stab 34F”
2′-OCF3
2′-O-
5′ and 3′-

Usually S

Methyl*
ends

CAP = any terminal cap, see for example FIG. 10.

All Stab 00-34 chemistries can comprise 3′-terminal thymidine (TT) residues

All Stab 00-34 chemistries typically comprise about 21 nucleotides, but can vary as described herein.

S = sense strand

AS = antisense strand

*Stab 23 has a single ribonucleotide adjacent to 3′-CAP

*Stab 24 and Stab 28 have a single ribonucleotide at 5′-terminus

*Stab 25, Stab 26, and Stab 27 have three ribonucleotides at 5′-terminus

*Stab 29, Stab 30, Stab 31, Stab 33, and Stab 34 any purine at first three nucleotide positions from 5′-terminus are ribonucleotides

p = phosphorothioate linkage

TABLE V

Reagent
Equivalents
Amount
Wait Time* DNA
Wait Time* 2′-O-methyl
Wait Time*RNA

A. 2.5 μmol Synthesis Cycle ABI 394 Instrument

Phosphoramidites
6.5
163
μL
45
sec
2.5
min
7.5
min

S-Ethyl Tetrazole
23.8
238
μL
45
sec
2.5
min
7.5
min

Acetic Anhydride
100
233
μL
5
sec
5
sec
5
sec

N-Methyl
186
233
μL
5
sec
5
sec
5
sec

Imidazole

TCA
176
2.3
mL
21
sec
21
sec
21
sec

Iodine
11.2
1.7
mL
45
sec
45
sec
45
sec

Beaucage
12.9
645
μL
100
sec
300
sec
300
sec

Acetonitrile
NA
6.67
mL
NA
NA
NA

B. 0.2 μmol Synthesis Cycle ABI 394 Instrument

Phosphoramidites
15
31
μL
45
sec
233
sec
465
sec

S-Ethyl Tetrazole
38.7
31
μL
45
sec
233
min
465
sec

Acetic Anhydride
655
124
μL
5
sec
5
sec
5
sec

N-Methyl
1245
124
μL
5
sec
5
sec
5
sec

Imidazole

TCA
700
732
μL
10
sec
10
sec
10
sec

Iodine
20.6
244
μL
15
sec
15
sec
15
sec

Beaucage
7.7
232
μL
100
sec
300
sec
300
sec

Acetonitrile
NA
2.64
mL
NA
NA
NA

C. 0.2 μmol Synthesis Cycle 96 well Instrument

Equivalents: DNA/
Amount: DNA/2′-O-

Wait Time* 2′-O-

Reagent
2′-O-methyl/Ribo
methyl/Ribo
Wait Time* DNA
methyl
Wait Time* Ribo

Phosphoramidites
22/33/66
40/60/120
μL
60
sec
180
sec
360
sec

S-Ethyl Tetrazole
70/105/210
40/60/120
μL
60
sec
180
min
360
sec

Acetic Anhydride
265/265/265
50/50/50
μL
10
sec
10
sec
10
sec

N-Methyl
502/502/502
50/50/50
μL
10
sec
10
sec
10
sec

Imidazole

TCA
238/475/475
250/500/500
μL
15
sec
15
sec
15
sec

Iodine
6.8/6.8/6.8
80/80/80
μL
30
sec
30
sec
30
sec

Beaucage
34/51/51
80/120/120

100
sec
200
sec
200
sec

Acetonitrile
NA
1150/1150/1150
μL
NA
NA
NA

Wait time does not include contact time during delivery.

Tandem synthesis utilizes double coupling of linker molecule

Number	Date	Country
60358580	Feb 2002	US
60358580	Feb 2002	US
60363124	Mar 2002	US
60363124	Mar 2002	US
60386782	Jun 2002	US
60386782	Jun 2002	US
60406784	Aug 2002	US
60406784	Aug 2002	US
60408378	Sep 2002	US
60408378	Sep 2002	US
60409293	Sep 2002	US
60409293	Sep 2002	US
60440129	Jan 2003	US
60440129	Jan 2003	US
60292217	May 2001	US
60362016	Mar 2002	US
60306883	Jul 2001	US
60311865	Aug 2001	US
60543480	Feb 2004	US

	Number	Date	Country
Parent	11098303	Apr 2005	US
Child	11217936	Sep 2005	US
Parent	10923536	Aug 2004	US
Child	11098303	Apr 2005	US
Parent	PCT/US04/16390	May 2004	US
Child	10923536	Aug 2004	US
Parent	10826966	Apr 2004	US
Child	PCT/US04/16390	May 2004	US
Parent	10757803	Jan 2004	US
Child	10826966	Apr 2004	US
Parent	10720448	Nov 2003	US
Child	10757803	Jan 2004	US
Parent	10693059	Oct 2003	US
Child	10720448	Nov 2003	US
Parent	10444853	May 2003	US
Child	10693059	Oct 2003	US
Parent	PCT/US03/05346	Feb 2003	US
Child	10444853	May 2003	US
Parent	PCT/US03/05028	Feb 2003	US
Child	10444853	May 2003	US
Parent	PCT/US04/13456	Apr 2004	US
Child	11217936	Sep 2005	US
Parent	10780447	Feb 2004	US
Child	PCT/US04/13456	Apr 2004	US
Parent	10427160	Apr 2003	US
Child	10780447	Feb 2004	US
Parent	PCT/US02/15876	May 2002	US
Child	10427160	Apr 2003	US
Parent	10727780	Dec 2003	US
Child	11217936	Sep 2005	US
Parent	PCT/US05/04270	Feb 2005	US
Child	11217936	Sep 2005	US

RNA interference mediated inhibition of histone deacetylase (HDAC) gene expression using short interfering nucleic acid (siNA)

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Abstract

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Provisional Applications (19)

Continuation in Parts (16)