NANOPARTICLE FORMULATIONS FOR DELIVERY OF NUCLEIC ACID COMPLEXES

FIELD OF THE DISCLOSURE

The disclosure relates to particle formulations, e.g., nanoparticle formulations, as well as methods of using such formulations for delivering oligonucleotides and/or synthetic RNA.

BACKGROUND OF THE DISCLOSURE

A considerable portion of human diseases can be treated by selectively altering protein and/or RNA levels of disease-associated transcription units (noncoding RNAs, protein-coding RNAs or other regulatory coding or noncoding genomic regions). Methods for inhibiting the expression of genes are known in the art and include, for example, antisense, RNAi and miRNA mediated approaches. Such methods may involve blocking translation of mRNAs or causing degradation of target RNAs. However, limited approaches are available for increasing the expression of genes. Furthermore, there is a need for effective formulation and delivery of agents useful for increasing the expression of genes in a subject.

SUMMARY OF THE DISCLOSURE

Aspects of the disclosure described herein relate to formulations and methods for delivery of a stabilizing oligonucleotide that is useful for modulating nucleic acids. In some embodiments, formulations and methods provided herein are useful for protecting RNAs (e.g., RNA transcripts) from degradation (e.g., exonuclease mediated degradation). In some embodiments, the protected RNAs are present outside of cells. In some embodiments, the protected RNAs are present in cells. In some embodiments, formulations and methods are provided that are useful for posttranscriptionally altering protein and/or RNA levels in a targeted manner. In some embodiments, methods disclosed herein involve reducing or preventing degradation or processing of targeted RNAs thereby elevating steady state levels of the targeted RNAs, e.g., in cells. In some embodiments, methods disclosed herein may also or alternatively involve increasing translation or increasing transcription of targeted RNAs, thereby elevating levels of RNA and/or protein levels in a targeted manner. In some embodiments, formulations and methods provided herein are useful for delivering a stabilizing oligonucleotide to a cell or tissue of interest.

Aspects of the disclosure relate to a recognition that certain RNA degradation is mediated by exonucleases. In some embodiments, exonucleases may destroy RNA from its 3′ end and/or 5′ end. As used herein, the term “stabilizing oligonucleotide” refers to an oligonucleotide (oligo) that hybridizes with RNA at or near one or both ends. Without wishing to be bound by theory, in some embodiments, it is believed that one or both ends of RNA can be protected from exonuclease enzyme activity by contacting the RNA with such stabilizing oligonucleotides, thereby increasing stability and/or levels of the RNA. The ability to increase stability and/or levels of a RNA by targeting the RNA at or near one or both ends, as disclosed herein, is surprising in part because of the presence of endonucleases (e.g., in cells) capable of destroying the RNA through internal cleavage. Moreover, in some embodiments, it is surprising that a 5′ targeting oligonucleotide is effective alone (e.g., not in combination with a 3′ targeting oligonucleotide or in the context of a pseudocircularization oligonucleotide) at stabilizing RNAs or increasing RNA levels because in cells, for example, 3′ end processing exonucleases may be dominant (e.g., compared with 5′ end processing exonucleases). However, in some embodiments, 3′ targeting oligonucleotides are used in combination with 5′ targeting oligonucleotides, or alone, to stabilize a target RNA.

In some embodiments, where a targeted RNA is protein-coding, increases in steady state levels of the RNA result in concomitant increases in levels of the encoded protein. Thus, in some embodiments, stabilizing oligonucleotides (including 5′-targeting, 3′-targeting and pseudocircularization oligonucleotides) are provided herein that when delivered to cells increase protein levels of target RNAs. In some embodiments, not only are target RNA levels increased but the resulting translation products are also increased. In some embodiments, this result is surprising in part because of an understanding that for translation to occur ribosomal machinery requires access to certain regions of the RNA (e.g., the 5′ cap region, start codon, etc.) to facilitate translation.

In some embodiments, where the targeted RNA is non-coding, increases in steady state levels of the non-coding RNA result in concomitant increases activity associated with the non-coding RNA. For example, in instances where the non-coding RNA is an miRNA, increases in steady state levels of the miRNA may result in increased degradation of mRNAs targeted by the miRNA.

In some embodiments, stabilizing oligonucleotides are provided with chemistries suitable for delivery, hybridization and stability within cells to target and stabilize RNA transcripts. Furthermore, in some embodiments, stabilizing oligonucleotide chemistries are provided that are useful for controlling the pharmacokinetics, biodistribution, bioavailability and/or efficacy of the oligonucleotides.

In some aspects of the disclosure, methods are provided for stabilizing a synthetic RNA (e.g., a synthetic RNA that is to be delivered to a cell). In some embodiments, the methods involve contacting a synthetic RNA with one or more stabilizing oligonucleotides that bind to a 5′ region of the synthetic RNA and a 3′ region of the synthetic RNA and that when bound to the synthetic RNA form a circularized product with the synthetic RNA. In some embodiments, the synthetic RNA is contacted with the one or more stabilizing oligonucleotides outside of a cell. Aspects of the disclosure relate to a formulation comprising: a single stranded synthetic nucleic acid, one or more stabilizing oligonucleotides complementary with the single stranded synthetic nucleic acid, and a particle. In some embodiments, the single stranded synthetic nucleic acid is a synthetic RNA.

In other aspects of the disclosure, methods are provided for stabilizing a nucleic acid in a cell (e.g., a messenger RNA present in a cell). In some embodiments, the methods involve contacting a messenger RNA in a cell with one or more stabilizing oligonucleotides that bind to a 5′ region of the RNA and a 3′ region of the messenger RNA and that when bound to the messenger RNA form a circularized product with the messenger RNA. Accordingly, aspects of the disclosure also relate to a formulation comprising: one or more stabilizing oligonucleotides, and a particle. In some embodiments, the particle is a nanoparticle. In some embodiments, the nanoparticle is a lipid nanoparticle.

In some embodiments, the particle is a nanoparticle. In some embodiments, the nanoparticle is a lipid nanoparticle.

In some embodiments, the lipid nanoparticle includes: (a) one or more cationic lipids, (b) one or more non-cationic lipids, (c) one or more conjugated lipids that inhibits aggregation of particles, or a combination thereof. In some embodiments, the lipid nanoparticle includes (a) one or more cationic lipids. In some embodiments, the lipid nanoparticle includes (a) one or more cationic lipid and (b) one or more non-cationic lipids. In some embodiments, the lipid nanoparticle includes (a) one or more cationic lipids, (b) one or more non-cationic lipids, and (c) one or more conjugated lipids that inhibit aggregation of particle.

In some embodiments, the lipid nanoparticle includes a cationic lipid selected from N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N—(I-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N—(I-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA), N,N-dimethyl-2,3-dioleyloxy)propylamine (DODMA), 1,2-DiLinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-Dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), 1,2-Dilinoleylcarbamoyloxy-3-dimethylaminopropane (DLin-C-DAP), 1,2-Dilinoleyoxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1,2-Dilinoleyoxy-3-morpholinopropane (DLin-MA), 1,2-Dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2-Dilinoleylthio-3-dimethylaminopropane (DLin-S-DMA), 1-Linoleoyl-2-linoleyloxy-3-dimethylaminopropane (DLin-2-DMAP), 1,2-Dilinoleyloxy-3-trimethylaminopropane chloride salt (DLin-TMA.Cl), 1,2-Dilinoleoyl-3-trimethylaminopropane chloride salt (DLin-TAP.Cl), 1,2-Dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), or 3-(N,N-Dilinoleylamino)-1,2-propanediol (DLinAP), 3-(N,N-Dioleylamino)-1,2-propanedio (DOAP), 1,2-Dilinoleyloxo-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), 1,2-Dilinolenyloxy-N,N-dimethylaminopropane (DLinDMA), 2,2-Dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA) or analogs thereof, (3aR,5s,6aS)—N,N-dimethyl-2,2-di((9Z,12Z)-octadeca-9, 12-dienyl)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-5-amine (ALN100), (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (MC3), 1, 1′-(2-(4-(2-((2-(bis(2-hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amino)ethyl)piperazin-1-yl)ethylazanediyl)didodecan-2-ol (Tech G1), 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane, β-L-arginyl-2, 3-L-diaminopropionic acid-N-palmityl-N-oleylamide trihydrochloride, N′,N′-dioctadecyl-N-4, 8-diaza-10-aminodecanoylglycine amide[71], 1,2-dilinoleyloxy-3-dimethylaminopropane, DLin-KC2-DMA, amino lipid 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA, 1), 1,2-distearloxy-/V,N-dimethylaminopropane (DSDMA), dilinoleylmethyl-4-dimethylaminobutyrate (DLin-MC3-DMA), DLin-D-DMA, C12-200, 98N12-5, (20Z,23Z)—N,N-dimethylnonacosa-20,23-dien-10-amine, (17Z,20Z)—N,N-dimemylhexacosa-17,20-dien-9-amine, (1Z,19Z)—N5N-dimethylpentacosa-16,19-dien-8-amine, (13Z,16Z)—N,N-dimethyldocosa-13,16-dien-5-amine, (12Z,15Z)—N,N-dimethylhenicosa-12,15-dien-4-amine, (14Z,17Z)—N,N-dimethyltricosa-14,17-dien-6-amine, (15Z,18Z)—N,N-dimethyltetracosa-15,18-dien-7-amine, (18Z,21Z)—N,N-dimethylheptacosa-18,21-dien-10-amine, (15Z,18Z)—N,N-dimethyltetracosa-15,18-dien-5-amine, (14Z,17Z)—N,N-dimethyltricosa-14,17-dien-4-amine, (19Z,22Z)—N,N-dimethyloctacosa-19,22-dien-9-amine, (18Z,21Z)—N,N-dimethylheptacosa-18,21-dien-8-amine, (17Z,20Z)—N,N-dimethylhexacosa-17,20-dien-7-amine, (16Z,19Z)—N,N-dimethylpentacosa-16,19-dien-6-amine, (22Z,25Z)—N,N-dimethylhentriaconta-22,25-dien-10-amine, (21Z,24Z)—N,N-dimethyltriaconta-21,24-dien-9-amine, (18Z)—N,N-dimethylheptacos-18-en-10-amine, (17Z)—N,N-dimethylhexacos-17-en-9-amine, (19Z,22Z)—N,N-dimethyloctacosa-19,22-dien-7-amine, N,N-dimethylheptacosan-10-amine, (20Z,23Z)—N-ethyl-N-methylnonacosa-20,23-dien-10-amine, 1-[(11Z,14Z)-1-nonylicosa-11,14-dien-1-yl]pyrrolidine, (20Z)—N,N-dimethylheptacos-20-en-10-amine, (15Z)—N,N-dimethyleptacos-15-en-10-amine, (14Z)—N,N-dimethylnonacos-14-en-10-amine, (17Z)—N,N-dimethylnonacos-17-en-10-amine, (24Z)—N,N-dimethyltritriacont-24-en-10-amine, (20Z)—N,N-dimethylnonacos-20-en-10-amine, (22Z)—N,N-dimethylhentriacont-22-en-10-amine, (16Z)—N,N-dimethylpentacos-16-en-8-amine, (12Z,15Z)—N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine, (13Z,16Z)—N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]eptadecan-8-amine, 1-[(1S,2R)-2-hexylcyclopropyl]-N,N-dimethylnonadecan-10-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]nonadecan-10-amine, N,N-dimethyl-21-[(1 S,2R)-2-octylcyclopropyl]henicosan-10-amine,N,N-dimeth-yl-1-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]methyl}cyclopropyl]nonadecan-10-amine,N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]hexadecan-8-amine, N,N-dimethyl-[(1R,2S)-2-undecylcyclopropyl]tetradecan-5-amine, N,N-dimethyl-3-{7-[(1S,2R)-2-octylcyclopropyl]heptyl}dodecan-i-amine, 1-[(1R,2S)-2-heptylcyclopropyl]-N,N-dimethyloctadecan-9-amine, 1-[(1S,2R)-2-decylcyclopropyl]-N,N-dimethylpentadecan-6-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]pentadecan-8-amine, R—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-(octyloxy)propa-n-2-amine, S—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-(octy-loxy)propan-2-amine, 1-{2-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-1-Roctyloxy)methyl]ethyl}pyrro-lidine, (2S)—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-[(5Z)-oct-5-en-1-yloxy]propan-2-amine, 1-{2-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-1-[(octyloxy)methyl]ethyl}azet-idine, (2S)-1-(hexyloxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-ylo-xy]propan-2-amine, (2S)-1-(heptyloxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]pr-opan-2-amine, N,N-dimethyl-1-(nonyloxy)-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine, N,N-dimethyl-1-[(9Z)-octadec-9-en-1-yloxy]-3-(octyloxy)propan-2-am-ine; (2S)—N,N-dimethyl-1-[(6Z,9Z,12Z)-octadeca-6,9,12-trien-1-yloxy]-3-(o-ctyloxy)propan-2-amine, (2 S)-1-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethyl-3-(pentyloxy)propa-n-2-amine, (2S)-1-(hexyloxy)-3-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-di-methylpropan-2-amine, 1-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, 1-[(13Z,16Z)-docosa-13,16-dien-1-yloxy]-N,N-dimethyl-3-(octyloxy)pr-opan-2-amine, (2S)-1-[(13Z,16Z)-docosa-13,16-dien-1-yloxy]-3-(hexyloxy)-N,N-dimethylpro-pan-2-amine, (2S)-1-[(13Z)-docos-13-en-1-yloxy]-3-(hexyloxy)-N,N-dimethylpropan-2-amin-e, 1-[(13Z)-docos-13-en-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, 1-[(9Z)-hexadec-9-en-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, (2R)—N,N-dimethyl-H(1-metoyloctyl)oxy]-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine, (2R)-1-[(3,7-dimethyloctyl)oxy]-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-di-en-1-yloxy]propan-2-amine, N,N-dimethyl-1-(octyloxy)-3-({8-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]-methyl}cyclopropyl]octyl}oxy)propan-2-amine, N,N-dimethyl-1-{[8-(2-oclylcyclopropyl)octyl]oxy}-3-(octyloxy)propan-2-am-ine and (11E,20Z,23Z)—N,N-dimethylnonacosa-11,20,2-trien-10-amine, 5-carboxyspermylglycine dioctaoleoylamide (“DOGS”), dipalmitoylphosphatidylethanolamine 5-carboxyspermyl-amide (“DPPES”), 1,2-dimyristyloxypropyl-3-dimethyl-hydroxy ethyl ammonium bromide (DMRIE), DMRIE-HP, Lipofectamine (DOSPA), 3b-(N—(N′,N′-dimethylaminoethane)-carbamoyl)cholesterol (“DC-Choi”), N-(1,2-dimyhstyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide (“DMRIE”), 1,2-Dioleoyl-3-dimethylammonium-propane (“DODAP”), DMDMA cationic lipid-based transfection reagents TransIT-TKO, LIPOFECTIN, Lipofectamine, OLIGOFECTAMINE or DHARMAFECT, DSDMA, DODMA, DLinDMA, DLenDMA, gamma-DLenDMA, DLin-K-DMA, DLin-K-C2-DMA (also known as DLin-C2K-DMA, XTC2, and C2K), DLin-K-C3-DM A, DLin-K-C4-DMA, DLen-C2K-DMA, y-DLen-C2K-DMA, DLin-M-C2-DMA (also known as MC2), DLin-M-C3-DMA (also known as MC3) and (DLin-MP-DMA)(also known as 1-B11), or a mixture thereof.

In some embodiments, the lipid nanoparticle includes a non-cationic lipid, wherein the non-cationic lipid is an anionic lipid. In some embodiments, the lipid nanoparticle includes a non-cationic lipid, wherein the non-cationic lipid is a neutral lipid.

In some embodiments, the lipid nanoparticle includes a non-cationic lipid selected from distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), cholesterol, phosphatidylglycerols, cardiolipins, diacylphosphatidylserines, diacylphosphatidic acids, N-dodecanoyl phosphatidylethanolamines, N-succinyl phosphatidylethanolamines, N-glutarylphosphatidylethanolamines, lysylphosphatidylglycerols, palmitoyloleyolphosphatidylglycerol (POPG), and other anionic modifying groups joined to neutral lipids, or a mixture thereof.

In some embodiments, the anionic lipid is 1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol).

In some embodiments, the neutral lipid is 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine.

In some embodiments, the lipid nanoparticle includes a conjugated lipid that inhibits aggregation of particles. In some embodiments, the conjugated lipid is a PEG lipid. In some embodiments, the PEG lipid is a PEG-diacylglycerol (DAG), a PEG-dialkyloxypropyl (DAA), a PEG-phospholipid, a PEG-ceramide (Cer), or a mixture thereof. In some embodiments, PLGA is conjugated to a lipid-terminating PEG forming PLGA-DSPE-PEG. In some embodiments, a PEG lipid is selected from PEG-c-DOMG and 1,2-Dimyristoyl-sn-glycerol, methoxypolyethylene Glycol (PEG-DMG), 1,2-Distearoyl-sn-glycerol, methoxypolyethylene Glycol (PEG-DSG), PEG-c-DOMG, 1,2-Distearoyl-sn-glycerol, methoxypolyethylene glycol (PEG-DSG) 1,2-Dipalmitoyl-sn-glycerol, methoxypolyethylene glycol (PEG-DPG), PEG-lipid conjugates such as, e.g., PEG coupled to dialkyloxypropyls (e.g., PEG-DAA conjugates), PEG coupled to diacylglycerols (e.g., PEG-DAG conjugates), PEG coupled to cholesterol, PEG coupled to phosphatidylethanolamines, and PEG conjugated to ceramides, cationic PEG lipids, polyoxazoline (POZ)-lipid conjugates, polyamide oligomers (e.g., ATTA-lipid conjugates), and mixtures thereof. In some embodiments, the PEG is a PEG-dilauryloxypropyl (C12), a PEG-dimyristyloxypropyl (C14), a PEG-dipalmityloxypropyl (C16), a PEG-distearyloxypropyl (C18), PEG-c-DOMG, PEG-DMG, or a mixture thereof.

In some embodiments, the conjugated PEG lipid is coupled to the surface of the lipid nanoparticle. In some embodiments, the PEG lipid is coupled to the surface of the lipid nanoparticle by an oxime linkage. In some embodiments, the conjugated PEG lipid is susceptible to decomposition in an acidic environment.

In some embodiments, formulations are provided in which the lipid nanoparticle comprises more than one cationic lipid, more than one non-cationic lipid, more than one conjugated lipid, or a combination thereof.

In some embodiments, formulations are provided in which the one or more stabilizing oligonucleotides are substantially encapsulated within an aqueous interior of the lipid nanoparticle.

In some embodiments, the lipid nanoparticle is about 50 to 150 nm in diameter. In some embodiments, the lipid nanoparticle is about 20-50 nm in diameter. In some embodiments, the lipid nanoparticle is about 30 nm in diameter.

In some embodiments, formulations are provided in which the lipid to stabilizing oligonucleotide ratio (mass/mass ratio; w/w ratio) is from about 1:1 to about 50:1, from about 1:1 to about 25:1, from about 10:1 to about 14:1, from about 3:1 to about 15:1, from about 4:1 to about 10:1, from about 5:1 to about 9:1, or about 6:1 to about 9:1.

In some embodiments, formulations are provided in which the particle is a microsphere. In some embodiments, the microsphere comprises poly(lactic-co-glycolic acid) (PLGA). In some embodiments, the microsphere particle is about 4 and 20 m in diameter.

In some embodiments, formulations are provided in which the particle includes a polymer. In some embodiments, the polymer comprises a layer of a hydrogel or surgical sealant. In some embodiments, the polymer is PLGA, ethylene vinyl acetate, poloxamer, GELSITE®, or a combination thereof.

In some embodiments, formulations are provided which further include protamine or calcium phosphate. In some embodiments, formulations are provided which further include hyaluronic acid. In some embodiments, formulations are provided which further include polyglutamate.

In some embodiments, formulations are provided in which the particle comprises a lipoprotein or lipoprotein mimetic. In some embodiments, the lipoprotein is HDL, LDL, or a combination thereof.

In some embodiments, formulations are provided in which the particle comprises a lipidoid. In some embodiments, the lipidoid is penta[3-(1-laurylaminopropionyl)]-triethylenetetramine hydrochloride (TETA-5LAP; 98N12-5), C12-200, MD1, or a combination thereof.

In some embodiments, the stabilizing oligonucleotide is a modified oligonucleotide. In some embodiments, the modified oligonucleotide comprises a modified sugar moiety, a modified internucleoside linkage, or a modified nucleotide, or a combination thereof.

In some embodiments, the stabilizing oligonucleotide is a mixmer. In some embodiment, the stabilizing oligonucleotide is a morpholino.

In some embodiments, the synthetic RNA comprises a transcription start site.

In some embodiments, the one or more stabilizing oligonucleotides comprises an oligonucleotide of 8 to 50 nucleotides in length and comprising a region of complementarity that is complementary with at least 5 contiguous nucleotides of the synthetic RNA, wherein the nucleotide at the 3′-end of the region of complementarity is complementary with a nucleotide within 10 nucleotides of the transcription start site of the synthetic RNA, wherein the oligonucleotide comprises nucleotides linked by at least one modified internucleoside linkage or at least one bridged nucleotide.

In some embodiments, the one or more stabilizing oligonucleotides comprises an oligonucleotide comprising two regions of complementarity each of which is complementary with at least 5 contiguous nucleotides of the synthetic RNA, wherein the nucleotide at the 3′-end of the first region of complementary is complementary with a nucleotide within 100 nucleotides of the transcription start site of the synthetic RNA and wherein the second region of complementarity is complementary with a region of the synthetic RNA that ends within 300 nucleotides of the 3′-end of the RNA transcript.

In some embodiments, the one or more stabilizing oligonucleotides comprises an oligonucleotide comprising the general formula 5′-X₁—X₂-3′, wherein X₁comprises 5 to 20 nucleotides that have a region of complementarity that is complementary with at least 5 contiguous nucleotides of a synthetic RNA, wherein the nucleotide at the 3′-end of the region of complementary of X₁is complementary with the nucleotide at the transcription start site of the RNA transcript; and X₂comprises 1 to 20 nucleotides.

In some embodiments, the synthetic RNA has a 7-methylguanosine cap at its 5′-end. In some embodiments, the synthetic RNA has a 7-methylguanosine cap, and the nucleotide at the 3′-end of the region of complementary of X₁is complementary with the nucleotide of the synthetic RNA that is immediately internal to the 7-methylguanosine cap.

In some embodiments, at least the first nucleotide at the 5′-end of X₂is a pyrimidine complementary with guanine. In some embodiments, the second nucleotide at the 5′-end of X₂is a pyrimidine complementary with guanine. In some embodiments, X₂comprises the formula 5′-Y₁-Y₂—Y₃-3′, wherein X₂forms a stem-loop structure having a loop region comprising the nucleotides of Y₂and a stem region comprising at least two contiguous nucleotides of Y₁hybridized with at least two contiguous nucleotides of Y₃. In some embodiments, Y₁, Y₂and Y₃independently comprise 1 to 10 nucleotides. In some embodiments, Y₃comprises, at a position immediately following the 3′-end of the stem region, a pyrimidine complementary with guanine. In some embodiments, the pyrimidine complementary with guanine is cytosine.

In some embodiments, X₂comprises a region of complementarity that is complementary with at least 5 contiguous nucleotides of the synthetic RNA that do not overlap the region of the synthetic RNA that is complementary with the region of complementarity of X₁. In some embodiments, the region of complementarity of X₂is within 100 nucleotides of a polyadenylation junction of the synthetic RNA. In some embodiments, the region of complementarity of X₂is complementary with the synthetic RNA immediately adjacent to or overlapping the polyadenylation junction of the synthetic RNA. In some embodiments, X₂further comprises at least 2 consecutive pyrimidine nucleotides complementary with adenine nucleotides of the poly(A) tail of the synthetic RNA.

In some embodiments, the synthetic RNA is an mRNA, non-coding RNA, long non-coding RNA, miRNA, or snoRNA or any other suitable RNA.

In some embodiments, the synthetic RNA is an mRNA transcript, and wherein X₂comprises a region of complementarity that is complementary with at least 5 contiguous nucleotides in the 3′-UTR of the transcript.

In some embodiments, formulations are provided in which the one or more stabilizing oligonucleotides comprises an oligonucleotide of 10 to 50 nucleotides in length having a first region complementary with at least 5 consecutive nucleotides of the 5′-UTR of a synthetic RNA, and a second region complementary with at least 5 consecutive nucleotides of the 3′-UTR, poly(A) tail, or overlapping the polyadenylation junction of the synthetic RNA. In some embodiments, the first of the at least 5 consecutive nucleotides of the 5′-UTR of the stabilizing oligonucleotide is within 10 nucleotides of the 5′-methylguanosine cap of the synthetic RNA. In some embodiments, the second region of the stabilizing oligonucleotide is complementary with at least 5 consecutive nucleotides overlapping the polyadenylation junction.

In some embodiments, the stabilizing oligonucleotide further comprises 2-20 nucleotides that link the 5′ end of the first region with the 3′ end of the second region. In some embodiments, the stabilizing oligonucleotide further comprises 2-20 nucleotides that link the 3′ end of the first region with the 5′ end of the second region. In some embodiments, the oligonucleotide is 10 to 50 nucleotide in length. In some embodiments, the oligonucleotide is 9 to 20 nucleotide in length.

In some embodiments, formulations are provided in which the one or more stabilizing oligonucleotides comprises an oligonucleotide comprising the general formula 5′-X1-X2-3′, wherein X1 comprises 2 to 20 pyrimidine nucleotides that form base pairs with adenine; and X2 comprises a region of complementarity that is complementary with at least 3 contiguous nucleotides of a poly-adenylated synthetic RNA, wherein the nucleotide at the 5′-end of the region of complementary of X2 is complementary with the nucleotide of the synthetic RNA that is immediately internal to the poly-adenylation junction of the synthetic RNA.

In some embodiments, formulations are provided in which the one or more stabilizing oligonucleotides comprises: a first oligonucleotide having 5 to 25 nucleotides linked through internucleoside linkages, and a second oligonucleotide having 5 to 25 nucleotides linked through internucleoside linkages, wherein the first oligonucleotide is complementary with at least 5 consecutive nucleotides within 100 nucleotides of the 5′-end of a synthetic RNA and wherein the second oligonucleotide is complementary with at least 5 consecutive nucleotides within 100 nucleotides of the 3′-end of a synthetic RNA.

In some embodiments, the first oligonucleotide and second oligonucleotide are joined by a linker that is not an oligonucleotide having a sequence complementary with the synthetic RNA. In some embodiments, the linker is an oligonucleotide. In some embodiments, the linker is a polypeptide.

In some embodiments, the synthetic RNA encodes a protein. In some embodiments, the synthetic RNA comprises one or more modified nucleotides.

In some embodiments, the formulation comprises a first and a second stabilizing oligonucleotide.

In some embodiments, the first stabilizing oligonucleotide comprises a region of complementarity to a 5′ region of the synthetic RNA and the second stabilizing oligonucleotide comprises a region of complementarity to a 3′ region of the synthetic RNA. In some embodiments, the first stabilizing oligonucleotide is covalently linked with the second stabilizing oligonucleotide. In some embodiments, the first stabilizing oligonucleotide and second stabilizing oligonucleotide are covalently linked through an internucleoside linkage. In some embodiments, the first stabilizing oligonucleotide and second stabilizing oligonucleotide are covalently linked through an oligonucleotide. In some embodiments, the first stabilizing oligonucleotide and second stabilizing oligonucleotide are covalently linked through a linker.

In some embodiments, the synthetic RNA is circularized. In some embodiments, the synthetic RNA has a 7-methylguanosine cap at its 5′-end.

In some embodiments, the first stabilizing oligonucleotide comprises a region of complementarity that is complementary with the synthetic RNA at a position within 10 nucleotides of the first nucleotide at the 5′ end of the synthetic RNA.

In some embodiments, the synthetic RNA comprises a 5′-methylguanosine cap, and wherein the first stabilizing oligonucleotide comprises a region of complementarity that is complementary with the synthetic RNA at a position within 10 nucleotides of the nucleotide immediately internal to the 5′-methylguanosine cap.

In some embodiments, the second stabilizing oligonucleotide comprises a region of complementarity that is complementary with the synthetic RNA at a position within 250 nucleotides of the 3′ end of the synthetic RNA.

In some embodiments, the synthetic RNA comprises a 3′-poly(A) tail, and wherein the second stabilizing oligonucleotide comprises a region of complementarity that is complementary with the synthetic RNA at a position within 100 nucleotides of the polyadenylation junction of the synthetic RNA.

In some embodiments, the region of complementarity of the second stabilizing oligonucleotide is immediately adjacent to or overlapping the polyadenylation junction of the synthetic RNA.

In some embodiments, the synthetic RNA is an RNA transcript. In some embodiments, the synthetic RNA is a functional RNA. In some embodiments, the synthetic RNA is up to 10 kb in length.

In some embodiments, the synthetic RNA comprises one or more modified nucleotides. In some embodiments, the one or more modified nucleotides are selected from the group consisting of: 2′-amino-2′-deoxynucleotide, 2′-azido-2′-deoxynucleotide, 2′-fluoro-2′-deoxynucleotide, 2′-O-methyl-nucleotide, 2′ sugar super modifier, 2′-modified thermostability enhancer, 2′-fluoro-2′-deoxyadenosine-5′-triphosphate, 2′-fluoro-2′-deoxycytidine-5′-triphosphate, 2′-fluoro-2′-deoxyguanosine-5′-triphosphate, 2′-fluoro-2′-deoxyuridine-5′-triphosphate, 2′-O-methyladenosine-5′-triphosphate, 2′-O-methylcytidine-5′-triphosphate, 2′-O-methylguanosine-5′-triphosphate, 2′-O-methyluridine-5′-triphosphate, pseudouridine-5′-triphosphate, 2′-O-methylinosine-5′-triphosphate, 2′-amino-2′-deoxycytidine-5′-triphosphate, 2′-amino-2′-deoxyuridine-5′-triphosphate, 2′-azido-2′-deoxycytidine-5′-triphosphate, 2′-azido-2′-deoxyuridine-5′-triphosphate, 2′-O-methylpseudouridine-5′-triphosphate, 2′-O-methyl-5-methyluridine-5′-triphosphate, 2′-azido-2′-deoxyadenosine-5′-triphosphate, 2′-amino-2′-deoxyadenosine-5′-triphosphate, 2′-fluoro-thymidine-5′-triphosphate, 2′-azido-2′-deoxyguanosine-5′-triphosphate, 2′-amino-2′-deoxyguanosine-5′-triphosphate, and N4-methylcytidine-5′-triphosphate.

In some aspects of the disclosure, methods are provided for delivering a synthetic RNA to a cell, the method comprising delivering to a cell a formulation of the present disclosure. In some embodiments, the increase in the level of gene expression results from an increase in the level of a protein encoded by the synthetic RNA. In some embodiments, the increase in the level of protein encoded by the synthetic RNA is at least a 50% increase compared with an appropriate cell to which the formulation was not delivered. In some embodiments, the cell is in vitro. In some embodiments, the cell is in vivo.

In some embodiments, the one or more stabilizing oligonucleotides comprises an oligonucleotide comprising two regions of complementarity each of which is complementary with at least 5 contiguous nucleotides of an RNA transcript, wherein the nucleotide at the 3′-end of the first region of complementary is complementary with a nucleotide within 100 nucleotides of the transcription start site of the RNA transcript and wherein the second region of complementarity is complementary with a region of the RNA transcript that ends within 300 nucleotides of the 3′-end of the RNA transcript.

In some embodiments, the one or more stabilizing oligonucleotides comprises an oligonucleotide comprising the general formula 5′-X₁—X₂-3′, wherein X1 comprises 5 to 20 nucleotides that have a region of complementarity that is complementary with at least 5 contiguous nucleotides of an RNA transcript, wherein the nucleotide at the 3′-end of the region of complementary of X₁is complementary with the nucleotide at the transcription start site of the RNA transcript; and X₂comprises 1 to 20 nucleotides.

In some embodiments, the RNA transcript has a 7-methylguanosine cap at its 5′-end. In some embodiments, the RNA transcript has a 7-methylguanosine cap, and the nucleotide at the 3′-end of the region of complementary of X₁is complementary with the nucleotide of the RNA transcript that is immediately internal to the 7-methylguanosine cap.

In some embodiments, X₂comprises a region of complementarity that is complementary with at least 5 contiguous nucleotides of the RNA transcript that do not overlap the region of the RNA transcript that is complementary with the region of complementarity of X₁. In some embodiments, the region of complementarity of X₂is within 100 nucleotides of a polyadenylation junction of the RNA transcript. In some embodiments, the region of complementarity of X₂is complementary with the RNA transcript immediately adjacent to or overlapping the polyadenylation junction of the RNA transcript. In some embodiments, X₂further comprises at least 2 consecutive pyrimidine nucleotides complementary with adenine nucleotides of the poly(A) tail of the RNA transcript.

In some embodiments, the RNA transcript is an mRNA, non-coding RNA, long non-coding RNA, miRNA, or snoRNA or any other suitable RNA.

In some embodiments, the RNA transcript is an mRNA transcript, and wherein X₂comprises a region of complementarity that is complementary with at least 5 contiguous nucleotides in the 3′-UTR of the transcript.

In some embodiments, formulations are provided in which the one or more stabilizing oligonucleotides comprises an oligonucleotide of 10 to 50 nucleotides in length having a first region complementary with at least 5 consecutive nucleotides of the 5′-UTR of an mRNA transcript, and a second region complementary with at least 5 consecutive nucleotides of the 3′-UTR, poly(A) tail, or overlapping the polyadenylation junction of the mRNA transcript. In some embodiments, the first of the at least 5 consecutive nucleotides of the 5′-UTR of the stabilizing oligonucleotide is within 10 nucleotides of the 5′-methylguanosine cap of the mRNA transcript. In some embodiments, the second region of the stabilizing oligonucleotide is complementary with at least 5 consecutive nucleotides overlapping the polyadenylation junction.

In some embodiments, formulations are provided in which the one or more stabilizing oligonucleotides comprises an oligonucleotide comprising the general formula 5′-X1-X2-3′, wherein X1 comprises 2 to 20 pyrimidine nucleotides that form base pairs with adenine; and X2 comprises a region of complementarity that is complementary with at least 3 contiguous nucleotides of a poly-adenylated RNA transcript, wherein the nucleotide at the 5′-end of the region of complementary of X2 is complementary with the nucleotide of the RNA transcript that is immediately internal to the poly-adenylation junction of the RNA transcript.

In some embodiments, formulations are provided in which the one or more stabilizing oligonucleotides comprises: a first oligonucleotide having 5 to 25 nucleotides linked through internucleoside linkages, and a second oligonucleotide having 5 to 25 nucleotides linked through internucleoside linkages, wherein the first oligonucleotide is complementary with at least 5 consecutive nucleotides within 100 nucleotides of the 5′-end of an RNA transcript and wherein the second oligonucleotide is complementary with at least 5 consecutive nucleotides within 100 nucleotides of the 3′-end of an RNA transcript.

In some embodiments, the first oligonucleotide and second oligonucleotide are joined by a linker that is not an oligonucleotide having a sequence complementary with the RNA transcript. In some embodiments, the linker is an oligonucleotide. In some embodiments, the linker is a polypeptide.

In some aspects of the disclosure, methods are provided for increasing gene expression in a cell, the method comprising delivering to a cell a formulation of the present disclosure. In some embodiments, the increase in the level of gene expression results from an increase in the level of a protein encoded by an mRNA that is stabilized by the one or more stabilizing oligonucleotides. In some embodiments, the increase in the level of protein encoded by an mRNA is at least a 50% increase compared with an appropriate cell to which the formulation was not delivered. In some embodiments, the cell is in vitro. In some embodiments, the cell is in vivo.

In some aspects of the disclosure, methods are provided for treating a condition or disease associated with decreased levels of an RNA transcript in a subject, the method comprising administering a formulation of the present disclosure to the subject. In some embodiments, the formulation is administered via topical, oral, or parenteral administration. In some embodiments, the parental administration is intravenous, subcutaneous, intraperitoneal, intramuscular, intrathecal, or intraventricular administration.

In some aspects of the disclosure, kits are provided, which include a container housing a formulation of the present disclosure.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is an illustration depicting exemplary oligo designs for targeting 3′ RNA ends. The first example shows oligos complementary to the 3′ end of RNA, before the polyA-tail. The second example shows oligos complementary to the 3′ end of RNA with a 5′ T-stretch to hybridize to a polyA tail.

FIG. 2 is an illustration depicting exemplary oligos for targeting 5′ RNA ends. The first example shows oligos complementary to the 5′ end of RNA. The second example shows oligos complementary to the 5′ end of RNA, the oligo having 3′ overhang residues to create a RNA-oligo duplex with a recessed end. Overhang can include a combination of nucleotides including, but not limited to, C to potentially interact with a 5′ methylguanosine cap and stabilize the cap further.

FIG. 3A is an illustration depicting exemplary oligos for targeting 5′ RNA ends and exemplary oligos for targeting 5′ and 3′ RNA ends. The example shows oligos with loops to stabilize a 5′ RNA cap or oligos that bind to a 5′ and 3′ RNA end to create a pseudo-circularized RNA.

FIG. 3B is an illustration depicting exemplary oligo-mediated RNA pseudocircularization. The illustration shows an LNA mixmer oligo binding to the 5′ and 3′ regions of an exemplary RNA.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE DISCLOSURE

Formulations and methods disclosed herein are useful in a variety of different contexts in which it is desirable to protect RNAs from degradation, including protecting RNAs inside or outside of cells. In some embodiments, formulations and methods are provided that are useful for posttranscriptionally altering protein and/or RNA levels in cells in a targeted manner. For example, formulations and methods are provided that involve reducing or preventing degradation or processing of targeted RNAs thereby elevating steady state levels of the targeted RNAs. In some embodiments, the stability of an RNA is increased by protecting one or both ends (5′ or 3′ ends) of the RNA from exonuclease activity, thereby increasing stability of the RNA.

In some embodiments, methods of increasing gene expression are provided. As used herein the term, “gene expression” refers generally to the level or representation of a product of a gene in a cell, tissue or subject. It should be appreciated that a gene product may be an RNA transcript or a protein, for example. An RNA transcript may be protein coding. An RNA transcript may be non-protein coding, such as, for example, a long non-coding RNA, a long intergenic non-coding RNA, a non-coding RNA, an miRNA, a small nuclear RNA (snRNA), or other functional RNA. In some embodiments, formulations and methods for increasing gene expression may involve increasing stability of a RNA transcript, and thereby increasing levels of the RNA transcript in the cell. Formulations and methods for of increasing gene expression may alternatively or in addition involve increasing transcription or translation of RNAs. In some embodiments, other mechanisms of manipulating gene expression may be involved in methods disclosed herein.

In some embodiments, methods provided herein involve delivering to a cell one or more sequence specific oligonucleotides that hybridize with an RNA transcript at or near one or both ends, thereby protecting the RNA transcript from exonuclease mediated degradation. In embodiments where the targeted RNA transcript is protein-coding, increases in steady state levels of the RNA typically result in concomitant increases in levels of the encoded protein. In embodiments where the targeted RNA is non-coding, increases in steady state levels of the non-coding RNA typically result in concomitant increases in activity associated with the non-coding RNA.

In some embodiments, approaches disclosed herein based on regulating RNA levels and/or protein levels using oligonucleotides targeting RNA transcripts by mechanisms that increase RNA stability and/or translation efficiency may have several advantages over other types of oligos or compounds, such as oligonucleotides that alter transcription levels of target RNAs using cis or noncoding based mechanisms. For example, in some embodiments, lower concentrations of oligos may be used when targeting RNA transcripts in the cytoplasm as multiple copies of the target molecules exist. In contrast, in some embodiments, oligos that target transcriptional processes may need to saturate the cytoplasm and before entering nuclei and interacting with corresponding genomic regions, of which there are only one/two copies per cell, in many cases. In some embodiments, response times may be shorter for RNA transcript targeting because RNA copies need not to be synthesized transcriptionally. In some embodiments, a continuous dose response may be easier to achieve. In some embodiments, well defined RNA transcript sequences facilitate design of oligonucleotides that target such transcripts. In some embodiments, oligonucleotide design approaches provided herein, e.g., designs having sequence overhangs, loops, and other features facilitate high oligo specificity and sensitivity compared with other types of oligonucleotides, e.g., certain oligonucleotides that target transcriptional processes.

In some embodiments, methods provided herein involve use of oligonucleotides that stabilize a nucleic acid (e.g., an RNA) by hybridizing at a single stranded 5′ and/or 3′ region of the nucleic acid. In some embodiments, oligonucleotides that prevent or inhibit degradation of an nucleic acid by hybridizing with the nucleic acid may be referred to herein as “stabilizing oligonucleotides.” In some examples, such oligonucleotides hybridize with an RNA and prevent or inhibit exonuclease mediated degradation of the RNA. Inhibition of exonuclease mediated degradation includes, but is not limited to, reducing the extent of degradation of a particular RNA by exonucleases. For example, an exonuclease that processes only single stranded RNA may cleave a portion of the RNA up to a region where an oligonucleotide is hybridized with the RNA because the exonuclease cannot effectively process (e.g., pass through) the duplex region. Thus, in some embodiments, using an oligonucleotide that targets a particular region of an RNA makes it possible to control the extent of degradation of the RNA by exonucleases up to that region. For example, use of an oligonucleotide that hybridizes at an end of an RNA may reduce or eliminate degradation by an exonuclease that processes only single stranded RNAs from that end. For example, use of an oligonucleotide that hybridizes at the 5′ end of an RNA may reduce or eliminate degradation by an exonuclease that processes single stranded RNAs in a 5′ to 3′ direction. Similarly, use of an oligonucleotide that hybridizes at the 3′ end of an RNA may reduce or eliminate degradation by an exonuclease that processes single stranded RNAs in a 3′ to 5′ direction. In some embodiments, lower concentrations of an oligo may be used when the oligo hybridizes at both the 5′ and 3′ regions of the RNA. In some embodiments, an oligo that hybridizes at both the 5′ and 3′ regions of the RNA protects the 5′ and 3′ regions of the RNA from degradation (e.g., by an exonuclease). In some embodiments, an oligo that hybridizes at both the 5′ and 3′ regions of the RNA creates a pseudo-circular RNA (e.g., a circularized RNA with a region of the poly A tail that protrudes from the circle, see FIG. 3B). In some embodiments, a pseudo-circular RNA is translated at a higher efficiency than a non-pseudo-circular RNA.

In some embodiments, an oligonucleotide may be used that comprises multiple regions of complementarity with an RNA, such that at one region the oligonucleotide hybridizes at or near the 5′ end of the RNA and at another region it hybridizes at or near the 3′ end of the RNA, thereby preventing or inhibiting degradation of the RNA by exonucleases at both ends. In some embodiments, when an oligonucleotide hybridizes both at or near the 5′ end of an RNA and at or near the 3′ end of the RNA a circularized complex results that is protected from exonuclease mediated degradation. In some embodiments, when an oligonucleotide hybridizes both at or near the 5′ end of an mRNA and at or near the 3′ end of the mRNA, the circularized complex that results is protected from exonuclease mediated degradation and the mRNA in the complex retains its ability to be translated into a protein.

As used herein the term, “synthetic RNA” refers to a RNA produced through an in vitro transcription reaction or through artificial (non-natural) chemical synthesis. In some embodiments, a synthetic RNA is an RNA transcript. In some embodiments, a synthetic RNA encodes a protein. In some embodiments, the synthetic RNA is a functional RNA (e.g., a lncRNA, miRNA, etc.). In some embodiments, a synthetic RNA comprises one or more modified nucleotides. In some embodiments, a synthetic RNA is up to 0.5 kilobases (kb), 1 kb, 1.5 kb, 2 kb, 2.5 kb, 3 kb, 4 kb, 5 kb, 6 kb, 7 kb, 8 kb, 9 kb, 10 kb, 15 kb, 20 kb, 25 kb, 30 kb or more in length. In some embodiments, a synthetic RNA is in a range of 0.1 kb to 1 kb, 0.5 kb to 2 kb, 0.5 kb to 10 kb, 1 kb to 5 kb, 2 kb to 5 kb, 1 kb to 10 kb, 3 kb to 10 kb, 5 kb to 15 kb, or 1 kb to 30 kb in length.

As used herein, the term “RNA transcript” refers to an RNA that has been transcribed from a nucleic acid by a polymerase enzyme. An RNA transcript may be produced inside or outside of cells. For example, an RNA transcript may be produced from a DNA template encoding the RNA transcript using an in vitro transcription reaction that utilizes recombination or purified polymerase enzymes. An RNA transcript may also be produced from a DNA template (e.g., chromosomal gene, an expression vector) in a cell by an RNA polymerase (e.g., RNA polymerase I, II, or III). In some embodiments, the RNA transcript is a protein coding mRNA. In some embodiments, the RNA transcript is a non-coding RNA (e.g., a tRNA, rRNA, snoRNA, miRNA, ncRNA, long-noncoding RNA, shRNA). In some embodiments, RNA transcript is up to 0.5 kilobases (kb), 1 kb, 1.5 kb, 2 kb, 2.5 kb, 3 kb, 4 kb, 5 kb, 6 kb, 7 kb, 8 kb, 9 kb, 10 kb, 15 kb, 20 kb, 25 kb, 30 kb or more in length. In some embodiments, a RNA transcript is in a range of 0.1 kb to 1 kb, 0.5 kb to 2 kb, 0.5 kb to 10 kb, 1 kb to 5 kb, 2 kb to 5 kb, 1 kb to 10 kb, 3 kb to 10 kb, 5 kb to 15 kb, or 1 kb to 30 kb in length.

In some embodiments, the RNA transcript is capped post-transcriptionally, e.g., with a 7′-methylguanosine cap. In some embodiments, the 7′-methylguanosine is added to the RNA transcript by a guanylyltransferase during transcription (e.g., before the RNA transcript is 20-50 nucleotides long.) In some embodiments, the 7′-methylguanosine is linked to the first transcribed nucleotide through a 5′-5′ triphosphate bridge. In some embodiments, the nucleotide immediately internal to the cap is an adenosine that is N6 methylated. In some embodiments, the first and second nucleotides immediately internal to the cap of the RNA transcript are not 2′-O-methylated. In some embodiments, the first nucleotide immediately internal to the cap of the RNA transcript is 2′-O-methylated. In some embodiments, the second nucleotide immediately internal to the cap of the RNA transcript is 2′-O-methylated. In some embodiments, the first and second nucleotides immediately internal to the cap of the RNA transcript are 2′-O-methylated.

In some embodiments, the RNA transcript is a non-capped transcript (e.g., a transcript produced from a mitochondrial gene). In some embodiments, the RNA transcript is a nuclear RNA that was capped but that has been decapped. In some embodiments, decapping of an RNA is catalyzed by the decapping complex, which may be composes of Dcp1 and Dcp2, e.g., that may compete with eIF-4E to bind the cap. In some embodiments, the process of RNA decapping involves hydrolysis of the 5′ cap structure on the RNA exposing a 5′ monophosphate. In some embodiments, this 5′ monophosphate is a substrate for the exonuclease XRN1. Accordingly, in some embodiments, an oligonucleotide that targets the 5′ region of an RNA may be used to stabilize (or restore stability) to a decapped RNA, e.g., protecting it from degradation by an exonuclease such as XRN1.

In some embodiments, in vitro transcription (e.g., performed via a T7 RNA polymerase or other suitable polymerase) may be used to produce an RNA transcript. In some embodiments transcription may be carried out in the presence of anti-reverse cap analog (ARCA) (TriLink Cat. # N-7003). In some embodiments, transcription with ARCA results in insertion of a cap (e.g., a cap analog (mCAP)) on the RNA in a desirable orientation.

In some embodiments, transcription is performed in the presence of one or more modified nucleotides (e.g., pseudouridine, 5-methylcytosine, etc.), such that the modified nucleotides are incorporated into the RNA transcript. It should be appreciated that any suitable modified nucleotide may be used, including, but not limited to, modified nucleotides that reduced immune stimulation, enhance translation and increase nuclease stability. Non-limiting examples of modified nucleotides that may be used include: 2′-amino-2′-deoxynucleotide, 2′-azido-2′-deoxynucleotide, 2′-fluoro-2′-deoxynucleotide, 2′-O-methyl-nucleotide, 2′ sugar super modifier, 2′-modified thermostability enhancer, 2′-fluoro-2′-deoxyadenosine-5′-triphosphate, 2′-fluoro-2′-deoxycytidine-5′-triphosphate, 2′-fluoro-2′-deoxyguanosine-5′-triphosphate, 2′-fluoro-2′-deoxyuridine-5′-triphosphate, 2′-O-methyladenosine-5′-triphosphate, 2′-O-methylcytidine-5′-triphosphate, 2′-O-methylguanosine-5′-triphosphate, 2′-O-methyluridine-5′-triphosphate, pseudouridine-5′-triphosphate, 2′-O-methylinosine-5′-triphosphate, 2′-amino-2′-deoxycytidine-5′-triphosphate, 2′-amino-2′-deoxyuridine-5′-triphosphate, 2′-azido-2′-deoxycytidine-5′-triphosphate, 2′-azido-2′-deoxyuridine-5′-triphosphate, 2′-O-methylpseudouridine-5′-triphosphate, 2′-O-methyl-5-methyluridine-5′-triphosphate, 2′-azido-2′-deoxyadenosine-5′-triphosphate, 2′-amino-2′-deoxyadenosine-5′-triphosphate, 2′-fluoro-thymidine-5′-triphosphate, 2′-azido-2′-deoxyguanosine-5′-triphosphate, 2′-amino-2′-deoxyguanosine-5′-triphosphate, and N4-methylcytidine-5′-triphosphate. In one embodiment, RNA degradation or processing can be reduced/prevented to elevate steady state RNA and, at least for protein-coding transcripts, protein levels. In some embodiments, a majority of degradation of RNA transcripts is done by exonucleases. In such embodiments, these enzymes start destroying RNA from either their 3′ or 5′ ends. By protecting the ends of the RNA transcripts from exonuclease enzyme activity, for instance, by hybridization of sequence-specific blocking oligonucleotides with proper chemistries for proper delivery, hybridization and stability within cells, RNA stability may be increase, along with protein levels for protein-coding transcripts.

In some embodiments, for the 5′ end, oligonucleotides may be used that are fully/partly complementary to 10-20 nts of the RNA 5′ end. In some embodiments, such oligonucleotides may have overhangs to form a hairpin (e.g., the 3′ nucleotide of the oligonucleotide can be, but not limited to, a C to interact with the mRNA 5′ cap's G nucleoside) to protect the RNA 5′ cap. In some embodiments, all nucleotides of an oligonucleotide may be complementary to the 5′ end of an RNA transcript, with or without few nucleotide overhangs to create a blunt or recessed 5′RNA-oligo duplex. In some embodiments, for the 3′ end, oligonucleotides may be partly complementary to the last several nucleotides of the RNA 3′ end, and optionally may have a poly(T)-stretch to protect the poly(A) tail from complete degradation (for transcripts with a poly(A)-tail). In some embodiments, similar strategies can be employed for other RNA species with different 5′ and 3′ sequence composition and structure (such as transcripts containing 3′ poly(U) stretches or transcripts with alternate 5′ structures). In some embodiments, oligonucleotides as described herein, including, for example, oligonucleotides with overhangs, may have higher specificity and sensitivity to their target RNA end regions compared to oligonucleotides designed to be perfectly complementary to RNA sequences, because the overhangs provide a destabilizing effect on mismatch regions and prefer binding in regions that are at the 5′ or 3′ ends of the RNAs. In some embodiments, oligonucleotides that protect the very 3′ end of the poly(A) tail with a looping mechanism (e.g., TTTTTTTTTTGGTTTTCC, SEQ ID NO: 458). In some embodiments, this latter approach may nonspecifically target all protein-coding transcripts. However, in some embodiments, such oligonucleotides, may be useful in combination with other target-specific oligos.

In some embodiments, methods provided herein involve the use of an oligonucleotide that comprises a region of complementarity that is complementary with the RNA transcript at a position at or near the first transcribed nucleotide of the RNA transcript. In some embodiments, an oligonucleotide (e.g., an oligonucleotide that stabilizes an RNA transcript) comprises a region of complementarity that is complementary with the RNA transcript (e.g., with at least 5 contiguous nucleotides) at a position that begins within 100 nucleotides, within 50 nucleotides, within 30 nucleotides, within 20 nucleotides, within 10 nucleotides or within 5 nucleotides of the 5′-end of the transcript. In some embodiments, an oligonucleotide (e.g., an oligonucleotide that stabilizes an RNA transcript) comprises a region of complementarity that is complementary with the RNA transcript (e.g., with at least 5 contiguous nucleotides of the RNA transcript) at a position that begins at the 5′-end of the transcript. In some embodiments, an oligonucleotide (e.g., an oligonucleotide that stabilizes an RNA transcript) comprises a region of complementarity that is complementary with an RNA transcript at a position within a region of the 5′ untranslated region (5′ UTR) of the RNA transcript spanning from the transcript start site to 50, 100, 150, 200, 250, 500 or more nucleotides upstream from a translation start site (e.g., a start codon, AUG, arising in a Kozak sequence of the transcript).

In some embodiments, an RNA transcript is poly-adenylated. Polyadenylation refers to the post-transcriptional addition of a polyadenosine (poly(A)) tail to an RNA transcript. Both protein-coding and non-coding RNA transcripts may be polyadenylated. Poly(A) tails contain multiple adenosines linked together through internucleoside linkages. In some embodiments, a poly(A) tail may contain 10 to 50, 25 to 100, 50 to 200, 150 to 250 or more adenosines. In some embodiments, the process of polyadenlyation involves endonucleolytic cleavage of an RNA transcript at or near its 3′-end followed by one by one addition of multiple adenosines to the transcript by a polyadenylate polymerase, the first of which adenonsines is added to the transcript at the 3′ cleavage site. Thus, often a polyadenylated RNA transcript comprises transcribed nucleotides (and possibly edited nucleotides) linked together through internucleoside linkages that are linked at the 3′ end to a poly(A) tail. The location of the linkage between the transcribed nucleotides and poly(A) tail may be referred to herein as, a “polyadenylation junction.” In some embodiments, endonucleolytic cleavage may occur at any one of several possible sites in an RNA transcript. In such embodiments, the sites may be determined by sequence motifs in the RNA transcript that are recognized by endonuclease machinery, thereby guiding the position of cleavage by the machinery. Thus, in some embodiments, polyadenylation can produce different RNA transcripts from a single gene, e.g., RNA transcripts have different polyadenylation junctions. In some embodiments, length of a poly(A) tail may determine susceptibility of the RNA transcript to enzymatic degradation by exonucleases with 3′-5′ processing activity. In some embodiments, oligonucleotides that target an RNA transcript at or near its 3′ end target a region overlapping a polyadenylation junction. In some embodiments, such oligonucleotides may have at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more nucleotides that are complementary with the transcribed portion of the transcript (5′ to the junction). In some embodiments, it is advantageous to have a limited number of nucleotides (e.g., T, U) complementary to the polyA side of the junction. In some embodiments, having a limited number of nucleotides complementary to the polyA side of the junction it is advantageous because it reduces toxicity associated with cross hybridization of the oligonucleotide to the polyadenylation region of non-target RNAs in cells. In some embodiments, the oligonucleotide has only 1, 2, 3, 4, 5, or 6 nucleotides complementary to the poly A region.

In some embodiments, methods provided herein involve the use of an oligonucleotide that hybridizes with a target RNA transcript at or near its 3′ end and prevents or inhibits degradation of the RNA transcript by 3′-5′ exonucleases. For example, in some embodiments, RNA stabilization methods provided herein involve the use of an oligonucleotide that comprises a region of complementarity that is complementary with the RNA transcript at a position within 100 nucleotides, within 50 nucleotides, within 30 nucleotides, within 20 nucleotides, within 10 nucleotides, within 5 nucleotides of the last transcribed nucleotide of the RNA transcript. In a case where the RNA transcript is a polyadenylated transcript, the last transcribed nucleotide of the RNA transcript is the first nucleotide upstream of the polyadenylation junction. In some embodiments, RNA stabilization methods provided herein involve the use of an oligonucleotide that comprises a region of complementarity that is complementary with the RNA transcript at a position immediately adjacent to or overlapping the polyadenylation junction of the RNA transcript. In some embodiments, RNA stabilization methods provided herein involve the use of an oligonucleotide that comprises a region of complementarity that is complementary with the RNA transcript within the poly(A) tail.

Particle Formulation, Delivery, and Dosing

Nucleic acids, e.g., stabilizing oligonucleotides and/or synthetic RNAs, described herein can be formulated with a particle for administration to a subject for treating a condition associated with decreased levels of expression of gene or instability or low stability of an RNA transcript that results in decreased levels of expression of a gene (e.g., decreased protein levels or decreased levels of functional RNAs, such as miRNAs, snoRNAs, lncRNAs, etc.). It should be understood that the formulations, compositions and methods can be practiced with any of the oligonucleotides and/or synthetic RNAs disclosed herein.

Any of the formulations, excipients, vehicles, etc. disclosed herein may be adapted or used to facilitate delivery of nucleic acids, e.g., stabilizing oligonucleotides and/or synthetic RNAs (e.g., linear or circularized synthetic RNAs), to a cell. Formulations, excipients, vehicles, etc. disclosed herein may be adapted or used to facilitate delivery of a nucleic acid to a cell in vitro or in vivo. Nucleic acids, e.g., stabilizing oligonucleotides and/or synthetic RNAs, may be formulated with a particle as described herein. For example, nucleic acids may be formulated with a nanoparticle, poly(lactic-co-glycolic acid) (PLGA) microsphere, lipidoid, lipoplex, liposome, polymer, carbohydrate (including simple sugars), cationic lipid, a fibrin gel, a fibrin hydrogel, a fibrin glue, a fibrin sealant, fibrinogen, thrombin, rapidly eliminated lipid nanoparticles (reLNPs) and combinations thereof. In some embodiments, a nucleic acid, e.g., stabilizing oligonucleotides and/or synthetic RNAs (e.g., linear or circularized synthetic RNAs), may be delivered to a cell gymnotically. In some embodiments, nucleic acids, e.g., stabilizing oligonucleotides and/or synthetic RNAs, may be conjugated with factors that facilitate delivery to cells. In some embodiments, a nucleic acid, e.g., stabilizing oligonucleotide and/or synthetic RNA, is conjugated with a carbohydrate, such as GalNac, or other targeting moiety.

In some aspects, a particle of the present disclosure is a nanoparticle, e.g., lipid nanoparticle. In some embodiments, a lipid nanoparticle may include one or more cationic lipids, one or more non-cationic lipids, one or more conjugated lipids that inhibits aggregation of particles, or a combination thereof. Such lipids can be used alone or in combination. In some embodiments, a lipid nanoparticle comprises more than one cationic lipid, more than one non-cationic lipid, more than one conjugated lipid, or a combination thereof. Exemplary components that may be present in the formulations and methods of the present disclosure are described below.

In some embodiments, a particle of the present disclosure includes a cationic lipid. As used herein, the term “cationic lipid” refers to a positively charged lipid. In some embodiments, cationic lipids can have certain features including a head group, one or more hydrophobic tails, and a linker between the head group and the one or more tails. The head group may include an amine. Under certain conditions, the amine nitrogen can be a site of positive charge. The cationic lipid for use in accordance with the present disclosure may be any appropriate cationic lipid for particle formulation, e.g., nanoparticle formulation, e.g., lipid nanoparticle formulation. For example, without limitation, the cationic lipid may be N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N—(I-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N—(I-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA), N,N-dimethyl-2,3-dioleyloxy)propylamine (DODMA), 1,2-DiLinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-Dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), 1,2-Dilinoleylcarbamoyloxy-3-dimethylaminopropane (DLin-C-DAP), 1,2-Dilinoleyoxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1,2-Dilinoleyoxy-3-morpholinopropane (DLin-MA), 1,2-Dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2-Dilinoleylthio-3-dimethylaminopropane (DLin-S-DMA), 1-Linoleoyl-2-linoleyloxy-3-dimethylaminopropane (DLin-2-DMAP), 1,2-Dilinoleyloxy-3-trimethylaminopropane chloride salt (DLin-TMA.Cl), 1,2-Dilinoleoyl-3-trimethylaminopropane chloride salt (DLin-TAP.Cl), 1,2-Dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), or 3-(N,N-Dilinoleylamino)-1,2-propanediol (DLinAP), 3-(N,N-Dioleylamino)-1,2-propanedio (DOAP), 1,2-Dilinoleyloxo-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), 1,2-Dilinolenyloxy-N,N-dimethylaminopropane (DLinDMA), 2,2-Dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA) or analogs thereof, (3aR,5s,6aS)-N,N-dimethyl-2,2-di((9Z,12Z)-octadeca-9, 12-dienyl)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-5-amine (ALN100), (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (MC3), 1, 1′-(2-(4-(2-((2-(bis(2-hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amino)ethyl)piperazin-1-yl)ethylazanediyl)didodecan-2-ol (Tech G1), 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane, β-L-arginyl-2, 3-L-diaminopropionic acid-N-palmityl-N-oleylamide trihydrochloride, N′,N′-dioctadecyl-N-4, 8-diaza-10-aminodecanoylglycine amide[71], 1,2-dilinoleyloxy-3-dimethylaminopropane, DLin-KC2-DMA, amino lipid 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA, 1), 1,2-distearloxy-/V,N-dimethylaminopropane (DSDMA), dilinoleylmethyl-4-dimethylaminobutyrate (DLin-MC3-DMA), DLin-D-DMA, C12-200, 98N12-5, (20Z,23Z)—N,N-dimethylnonacosa-20,23-dien-10-amine, (17Z,20Z)—N,N-dimemylhexacosa-17,20-dien-9-amine, (1Z,19Z)—N5N-dimethylpentacosa-16,19-dien-8-amine, (13Z,16Z)—N,N-dimethyldocosa-13,16-dien-5-amine, (12Z,15Z)—N,N-dimethylhenicosa-12,15-dien-4-amine, (14Z,17Z)—N,N-dimethyltricosa-14,17-dien-6-amine, (15Z,18Z)—N,N-dimethyltetracosa-15,18-dien-7-amine, (18Z,21Z)—N,N-dimethylheptacosa-18,21-dien-10-amine, (15Z,18Z)—N,N-dimethyltetracosa-15,18-dien-5-amine, (14Z,17Z)—N,N-dimethyltricosa-14,17-dien-4-amine, (19Z,22Z)—N,N-dimethyloctacosa-19,22-dien-9-amine, (18Z,21Z)—N,N-dimethylheptacosa-18,21-dien-8-amine, (17Z,20Z)—N,N-dimethylhexacosa-17,20-dien-7-amine, (16Z,19Z)—N,N-dimethylpentacosa-16,19-dien-6-amine, (22Z,25Z)—N,N-dimethylhentriaconta-22,25-dien-10-amine, (21Z,24Z)—N,N-dimethyltriaconta-21,24-dien-9-amine, (18Z)—N,N-dimethylheptacos-18-en-10-amine, (17Z)—N,N-dimethylhexacos-17-en-9-amine, (19Z,22Z)—N,N-dimethyloctacosa-19,22-dien-7-amine, N,N-dimethylheptacosan-10-amine, (20Z,23Z)—N-ethyl-N-methylnonacosa-20,23-dien-10-amine, 1-[(11Z,14Z)-1-nonylicosa-11,14-dien-1-yl]pyrrolidine, (20Z)—N,N-dimethylheptacos-20-en-10-amine, (15Z)—N,N-dimethyleptacos-15-en-10-amine, (14Z)—N,N-dimethylnonacos-14-en-10-amine, (17Z)—N,N-dimethylnonacos-17-en-10-amine, (24Z)—N,N-dimethyltritriacont-24-en-10-amine, (20Z)—N,N-dimethylnonacos-20-en-10-amine, (22Z)—N,N-dimethylhentriacont-22-en-10-amine, (16Z)—N,N-dimethylpentacos-16-en-8-amine, (12Z,15Z)—N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine, (13Z,16Z)—N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]eptadecan-8-amine, 1-[(1S,2R)-2-hexylcyclopropyl]-N,N-dimethylnonadecan-10-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]nonadecan-10-amine, N,N-dimethyl-21-[(1S,2R)-2-octylcyclopropyl]henicosan-10-amine,N,N-dimeth-yl-1-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]methyl}cyclopropyl]nonadecan-10-amine,N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]hexadecan-8-amine, N,N-dimethyl-[(1R,2S)-2-undecylcyclopropyl]tetradecan-5-amine, N,N-dimethyl-3-{7-[(1 S,2R)-2-octylcyclopropyl]heptyl}dodecan-1-amine, 1-[(1R,2S)-2-heptylcyclopropyl]-N,N-dimethyloctadecan-9-amine, 1-[(1S,2R)-2-decylcyclopropyl]-N,N-dimethylpentadecan-6-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]pentadecan-8-amine, R—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-(octyloxy)propa-n-2-amine, S—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-(octy-loxy)propan-2-amine, 1-{2-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-1-Roctyloxy)methyl]ethyl}pyrro-lidine, (2S)—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-[(5Z)-oct-5-en-1-yloxy]propan-2-amine, 1-{2-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-1-[(octyloxy)methyl]ethyl}azet-idine, (2S)-1-(hexyloxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-ylo-xy]propan-2-amine, (2S)-1-(heptyloxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]pr-opan-2-amine, N,N-dimethyl-1-(nonyloxy)-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine, N,N-dimethyl-1-[(9Z)-octadec-9-en-1-yloxy]-3-(octyloxy)propan-2-am-ine; (2S)—N,N-dimethyl-1-[(6Z,9Z,12Z)-octadeca-6,9,12-trien-1-yloxy]-3-(o-ctyloxy)propan-2-amine, (2 S)-1-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethyl-3-(pentyloxy)propa-n-2-amine, (2S)-1-(hexyloxy)-3-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-di-methylpropan-2-amine, 1-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, 1-[(13Z,16Z)-docosa-13,16-dien-1-yloxy]-N,N-dimethyl-3-(octyloxy)pr-opan-2-amine, (2S)-1-[(13Z,16Z)-docosa-13,16-dien-1-yloxy]-3-(hexyloxy)-N,N-dimethylpro-pan-2-amine, (2S)-1-[(13Z)-docos-13-en-1-yloxy]-3-(hexyloxy)-N,N-dimethylpropan-2-amin-e, 1-[(13Z)-docos-13-en-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, 1-[(9Z)-hexadec-9-en-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, (2R)—N,N-dimethyl-H(1-metoyloctyl)oxy]-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine, (2R)-1-[(3,7-dimethyloctyl)oxy]-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-di-en-1-yloxy]propan-2-amine, N,N-dimethyl-1-(octyloxy)-3-({8-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]-methyl}cyclopropyl]octyl}oxy)propan-2-amine, N,N-dimethyl-1-{[8-(2-oclylcyclopropyl)octyl]oxy}-3-(octyloxy)propan-2-am-ine and (11E,20Z,23Z)—N,N-dimethylnonacosa-11,20,2-trien-10-amine, 5-carboxyspermylglycine dioctaoleoylamide (“DOGS”), dipalmitoylphosphatidylethanolamine 5-carboxyspermyl-amide (“DPPES”), 1,2-dimyristyloxypropyl-3-dimethyl-hydroxy ethyl ammonium bromide (DMRIE), DMRIE-HP, Lipofectamine (DOSPA), 3b-(N—(N′,N′-dimethylaminoethane)-carbamoyl)cholesterol (“DC-Choi”), N-(1,2-dimyhstyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide (“DMRIE”), 1,2-Dioleoyl-3-dimethylammonium-propane (“DODAP”), DMDMA, cationic lipid-based transfection reagents TransIT-TKO, LIPOFECTIN, Lipofectamine, OLIGOFECTAMINE or DHARMAFECT, DSDMA, DODMA, DLinDMA, DLenDMA, gamma-DLenDMA, DLin-K-DMA, DLin-K-C2-DMA (also known as DLin-C2K-DMA, XTC2, and C2K), DLin-K-C3-DM A, DLin-K-C4-DMA, DLen-C2K-DMA, y-DLen-C2K-DMA, DLin-M-C2-DMA (also known as MC2), DLin-M-C3-DMA (also known as MC3) and (DLin-MP-DMA)(also known as 1-B11) or a mixture thereof. In some embodiments, the cationic lipid may comprise from about 20 mol percent to about 50 mol percent or about 40 mol percent of the total lipid present in the particle.

The non-cationic lipid may be any appropriate non-cationic lipid for particle, e.g., nanoparticle, e.g., lipid nanoparticle, formulation. For example, the non-cationic lipid may be an anionic lipid or a neutral lipid. Anionic lipids suitable for use in particles of the present disclosure include, but are not limited to, phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacylphosphatidic acid, N-dodecanoyl phosphatidylethanoloamine, N-succinyl phosphatidylethanolamine, N-glutaryl phosphatidylethanolamine, lysylphosphatidylglycerol, and other anionic modifying groups joined to neutral lipids. In some embodiments, the anionic lipid is 1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol).

A neutral lipid generally refers to a lipid which exists either in an uncharged or neutral zwitterionic form at physiological pH. Such lipids include, for example diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, dihydrosphingomyelin, cephalin, and cerebrosides. The selection of neutral lipids for use in the particles described herein is generally guided by consideration of, e.g., liposome size and stability of the liposomes in the bloodstream. In some embodiments, the neutral lipid component is a lipid having two acyl groups, (e.g., diacylphosphatidylcholine and diacylphosphatidylethanolamine). Lipids having a variety of acyl chain groups of varying chain length and degree of saturation are available or may be isolated or synthesized by well-known techniques. In some embodiments, lipids contain saturated fatty acids with carbon chain lengths in the range of C₁₀to C₂₀. In some embodiments, lipids with mono or diunsaturated fatty acids with carbon chain lengths in the range of C₁₀to C₂₀may be used. Additionally, lipids having mixtures of saturated and unsaturated fatty acid chains can be used. For example, a neutral lipid for use in accordance with the present disclosure is DOPE, DSPC, POPC, DPPC or any related phosphatidylcholine. In some embodiments, a neutral lipid may be composed of sphingomyelin, dihydrosphingomyeline, or phospholipids with other head groups, such as serine and inositol. In some embodiments, the neutral lipid is 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine.

In some embodiments, exemplary non-cationic lipids include, but are not limited to, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), cholesterol, phosphatidylglycerols, cardiolipins, diacylphosphatidylserines, diacylphosphatidic acids, N-dodecanoyl phosphatidylethanolamines, N-succinyl phosphatidylethanolamines, N-glutarylphosphatidylethanolamines, lysylphosphatidylglycerols, palmitoyloleyolphosphatidylglycerol (POPG), and other anionic modifying groups joined to neutral lipids or a mixture thereof. In some embodiments, the anionic lipid is 1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol). In some embodiments, the neutral lipid is 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine.

Particles of the present disclosure may include a lipid that inhibits and/or reduces aggregation of particles during formation. Examples of lipids that reduce aggregation of particles during formation include polyethylene glycol (PEG)-modified lipids, monosialoganglioside Gm1, and polyamide oligomers (“PAO”) such as (described in U.S. Pat. No. 6,320,017). Other compounds with uncharged, hydrophilic, steric-barrier moieties, which prevent aggregation during formulation, like PEG, Gm1 or ATTA, can also be coupled to lipids for use as in accordance with the disclosure. Exemplary ATTA-lipids are described, e.g., in U.S. Pat. No. 6,320,017, and exemplary PEG-lipid conjugates are described, e.g., in U.S. Pat. Nos. 5,820,873, 5,534,499 and 5,885,613.

In some embodiments, the conjugated lipid is a PEG lipid. In some embodiments, the PEG lipid is a PEG-diacylglycerol (DAG), a PEG-dialkyloxypropyl (DAA), a PEG-phospholipid, a PEG-ceramide (Cer), or a mixture thereof. As a non-limiting example, PLGA may be conjugated to a lipid-terminating PEG forming PLGA-DSPE-PEG, PEG lipid is selected from PEG-c-DOMG and 1,2-Dimyristoyl-sn-glycerol, methoxypolyethylene Glycol (PEG-DMG), 1,2-Distearoyl-sn-glycerol, methoxypolyethylene Glycol (PEG-DSG), PEG-c-DOMG, 1,2-Distearoyl-sn-glycerol, methoxypolyethylene glycol (PEG-DSG) 1,2-Dipalmitoyl-sn-glycerol, methoxypolyethylene glycol (PEG-DPG), PEG-lipid conjugates such as, e.g., PEG coupled to dialkyloxypropyls (e.g., PEG-DAA conjugates), PEG coupled to diacylglycerols (e.g., PEG-DAG conjugates), PEG coupled to cholesterol, PEG coupled to phosphatidylethanolamines, and PEG conjugated to ceramides, cationic PEG lipids, polyoxazoline (POZ)-lipid conjugates, polyamide oligomers (e.g., ATTA-lipid conjugates), and mixtures thereof. In some embodiments, the PEG is a PEG-dilauryloxypropyl (C12), a PEG-dimyristyloxypropyl (C14), a PEG-dipalmityloxypropyl (C16), a PEG-distearyloxypropyl (C18), PEG-c-DOMG, PEG-DMG, or a mixture thereof.

In some embodiments, the conjugated lipid that prevents aggregation of particles may be from 0% to about 20%, e.g., about 1% to about 15%, e.g., about 2% of the total lipid present in the particle (by mole percent of lipids). In some embodiments, the nucleic acid-lipid particle further includes cholesterol at, e.g., about 10 mol percent to about 60 mol percent or about 48 mol percent of the total lipid present in the particle.

In some embodiments, the lipid nanoparticles of the present disclosure have a mean diameter of about 50 nm to about 150 nm, e.g., about 60 nm to about 130 nm, about 70 nm to about 110 nm, about 60 nm to about 80 nm. In some embodiments, the lipid nanoparticles of the present disclosure have a mean diameter of about 20-50 nm. In some embodiments, the lipid nanoparticles of the present disclosure have a mean diameter of about 30 nm. In some embodiments, the particle size is at least 30 nm, at least 40 nm, at least 50 nm, at least 60 nm, at least 70 nm, at least 80 nm, at least 90 nm, at least 100 nm, at least 110 nm, or at least 120 nm. In some embodiments, the particle size range is about at least 50 nm to about at least 110 nm, about at least 60 nm to about at least 100 nm, or about at least 50 nm to about at least 80 nm.

In some embodiments, nucleic acids, e.g., stabilizing oligonucleotide and/or synthetic RNA, are encapsulated in the particle of the present disclosure. In some embodiments, encapsulated nucleic acids are resistant in aqueous solution to degradation with a nuclease. In some embodiments, the nucleic acid, e.g., stabilizing oligonucleotide and/or synthetic RNA, encapsulated in the particles is not significantly degraded after exposure to serum or a nuclease assay that would significantly degrade free nucleic acid. Nuclease degradation of nucleic acids may be determined by an Oligreen® assay (Invitrogen Corporation, Carlsbad, Calif.), which is an ultra-sensitive fluorescent nucleic acid stain for quantitating oligonucleotides and single-stranded DNA in solution. In some embodiments, nucleic acids, e.g., stabilizing oligonucleotide and/or synthetic RNA, are at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 97% encapsulated in the particle.

In one embodiment, the lipid to drug ratio (mass/mass ratio; w/w ratio) (e.g., lipid to dsRNA ratio) will be in the range of from about 1:1 to about 50:1, from about 1:1 to about 25:1, from about 10:1 to about 14:1, from about 3:1 to about 15:1, from about 4:1 to about 10:1, from about 5:1 to about 9:1, or about 6:1 to about 9:1.

In some embodiments, a particle of the present disclosure comprises a polymer. In some embodiments, the polymer comprises a layer of a hydrogel or surgical sealant. In some embodiments, the polymer is PLGA, ethylene vinyl acetate, poloxamer, GELSITE®, or a combination thereof. In some embodiments, a particle of the present disclosure comprises a fibrin sealant.

In some embodiments, it is desirable to precondense the nucleic acid, e.g., stabilizing oligonucleotide and/or synthetic RNA, into the core of a particle described herein. In some embodiments, a heper cationic polymer, such as protamine is included in the formulation. In some embodiments, protamine interacts with nucleic acid to form a negatively charged compact core.

In some embodiments, calcium phosphate is included in a formulation described herein. In some embodiments, hyaluronic acid is included in a formulation described herein. In some embodiments, polyglutamate is included in a formulation described herein

In some embodiments, a particle of the disclosure comprises a lipoprotein or lipoprotein mimetic, or fragment thereof. In some embodiments, the lipoprotein is HDL, LDL, or a combination thereof.

In some aspects, a particle in accordance with the present disclosure comprises a lipidoid. The synthesis of lipidoids has been extensively described and formulations containing these compounds are particularly suited for delivery of modified nucleic acid molecules or mmRNA (see Mahon et al., Bioconjug Chem. 2010 21:1448-1454; Schroeder et al., J Intern Med. 2010 267:9-21; Akinc et al., Nat Biotechnol. 2008 26:561-569; Love et al., Proc Natl Acad Sci USA. 2010 107:1864-1869; Siegwart et al., Proc Natl Acad Sci USA. 2011 108:12996-3001; all of which are incorporated herein in their entireties). Complexes, micelles, liposomes or particles can be prepared containing a lipidoid and therefore, can result in an effective delivery of the nucleic acids, e.g., stabilizing oligonucleotides and/or synthetic RNAs, of the present disclosure. In some embodiments, the lipidoid is penta[3-(1-laurylaminopropionyl)]-triethylenetetramine hydrochloride (TETA-5LAP; 98N12-5), C12-200, MD1, or a combination thereof. The lipidoid formulations can include particles comprising, e.g., 3 or 4 or more components in addition to stabilizing oligonucleotide and/or synthetic RNA. As an example, formulations with certain lipidoids, include, but are not limited to, 98N12-5 and may contain 42% lipidoid, 48% cholesterol and 10% PEG (C14 alkyl chain length). As another example, formulations with certain lipidoids, include, but are not limited to, C12-200 and may contain 50% lipidoid, 10% disteroylphosphatidyl choline, 38.5% cholesterol, and 1.5% PEG-DMG.

In some embodiments, a formulation comprises a rapidly eliminated lipid nanoparticle (reLNP). In some embodiments, a reLNPcomprises a reLNP lipid, a fusogenic lipid, cholesterol and a PEG lipid at a molar ratio of 50:10:38.5:1.5 (reLNP lipid:fusogenic lipid:cholesterol:PEG lipid). The fusogenic lipid may be DSPC and the PEG lipid may be PEG-c-DOMG. The reLNP lipid may be DLin-DMA with an internal or terminal ester or DLin-MC3-DMA with an internal or terminal ester. In some embodiments, the total lipid to modified mRNA weight ratio is between 10:1 and 30:1.

The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient (e.g., an oligonucleotide or compound of the disclosure) which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration, e.g., intradermal or inhalation. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect, e.g. tumor regression.

Pharmaceutical formulations of this disclosure can be prepared according to any method known to the art for the manufacture of pharmaceuticals. Such formulations can contain sweetening agents, flavoring agents, coloring agents and preserving agents. A formulation can be admixtured with nontoxic pharmaceutically acceptable excipients which are suitable for manufacture. Formulations may comprise one or more diluents, emulsifiers, preservatives, buffers, excipients, etc. and may be provided in such forms as liquids, powders, emulsions, lyophilized powders, sprays, creams, lotions, controlled release formulations, tablets, pills, gels, on patches, in implants, etc.

A formulated nucleic acid, e.g., oligonucleotide and/or synthetic RNA, and particle composition can assume a variety of states. In some examples, the composition is at least partially crystalline, uniformly crystalline, and/or anhydrous (e.g., less than 80, 50, 30, 20, or 10% water). In another example, a nucleic acid, e.g., oligonucleotide and/or synthetic RNA, is in an aqueous phase, e.g., in a solution that includes water. The aqueous phase or the crystalline compositions can, e.g., be incorporated into a delivery vehicle, e.g., a liposome (particularly for the aqueous phase) or a particle (e.g., a microparticle as can be appropriate for a crystalline composition) as described herein. Generally, a nucleic acid, e.g., oligonucleotide and/or synthetic RNA, composition is formulated in a manner that is compatible with the intended method of administration.

In some embodiments, the composition is prepared by at least one of the following methods: spray drying, lyophilization, vacuum drying, evaporation, fluid bed drying, or a combination of these techniques; or sonication with a lipid, freeze-drying, condensation and other self-assembly.

A nucleic acid, e.g., oligonucleotide and/or synthetic RNA, preparation can be formulated or administered (together or separately) in combination with another agent, e.g., another therapeutic agent or an agent that stabilizes an oligonucleotide, e.g., a protein that complexes with oligonucleotide. Still other agents include chelators, e.g., EDTA (e.g., to remove divalent cations such as Mg²⁺), salts, RNAse inhibitors (e.g., a broad specificity RNAse inhibitor such as RNAsin) and so forth.

In one embodiment, an formulation comprising an oligonucleotide includes another oligonucleotide, e.g., a second oligonucleotide that modulates expression of a second gene or a second oligonucleotide that modulates expression of the first gene. Still other preparation can include at least 3, 5, ten, twenty, fifty, or a hundred or more different oligonucleotide species. Such oligonucleotides can mediated gene expression with respect to a similar number of different genes. In one embodiment, an oligonucleotide preparation includes at least a second therapeutic agent (e.g., an agent other than an oligonucleotide).

Ligand Modified Particles

In some aspects, a particle of the present disclosure may be conjugated to a targeting ligand that targets the particle to a particular ligand binding molecule, e.g., a ligand binding molecule present in a particular tissue or on the surface of a particular cell. Any suitable ligand may be used, including protein, lipid, polynucleotide, polysaccharide, or other molecules. In some embodiments, without limitation, the ligand is an antibody or antigen binding fragment thereof, a small molecule, a drug, a chemical, an ion, an aptamer, an oligonucleotide, a single stranded oligonucleotide, DNA, RNA, a peptide, a protein, a neuropeptide, or a neurotransmitter.

In some embodiments, a targeting ligand may be conjugated to a particle directly, e.g., through a direct covalent linkage. However, in some embodiments, a targeting ligand may be conjugated to a particle indirectly, e.g., via another molecule, such as a linker or adaptor molecule. As used herein, the term “particle linker” refers to any linker that is conjugated to a particle. In some embodiments, a targeting ligand is conjugated to a particle (e.g., a lipid nanoparticle) via a particle linker. Any suitable particle linker may be used. In some embodiments, a particle linker is conjugated to a targeting ligand. In some embodiments, the particle linker is a hetero-bifunctional molecule, such as polyethylene glycol (PEG) (e.g., as described herein and/or in U.S. Pat. Nos. 5,820,873, 5,534,499 and/or 5,885,613, incorporated by reference herein, in their entireties). In some embodiments, the particle linker is a monosialoganglioside (Gm1/GM1/GM-1), a polyamide oligomer (e.g., PAO e.g., ATTA, e.g., N-(w-azido-octa-(14′-amino-3′,6′,9′,12′-tetraoxatetradecanoyl))-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (ATTA-DPSE)) (e.g., as described in U.S. Pat. Nos. 6,320,017 and/or 6,586,559, incorporated by reference herein, in their entireties). GM1 is a member of the ganglio series of gangliosides which contain one sialic acid residue. GM1 impacts neuronal plasticity and repair mechanisms and the release of neurotrophins in the brain. Polyamides are cell permeable, synthetic oligomers. In some embodiments, the particle is conjugated to a mixture of particle linkers. In some embodiments, the particle is conjugated to a targeting ligand. In some embodiments, the particle is conjugated to a mixture of targeting ligands. In some embodiments, the particle is conjugated to a mixture of particle linkers and targeting ligands.

In some embodiments the particle is a nanoparticle. In some embodiments, the nanoparticle is a polymeric nanoparticle, micelle, dendrimer, or liposome as described herein.

In some aspects, a nucleic acid complex (e.g., containing a stabilizing oligonucleotide and synthetic mRNA) or an oligonucleotide (e.g., stabilizing oligonucleotide) formulation disclosed herein can be targeted to a specific cell or tissue type of interest via a ligand conjugated to a particle or via a ligand conjugated to a particle linker, which is conjugated to the particle, wherein the ligand targets a receptor of the cell or tissue of interest. For example, in some embodiments, a ligand can be mannose, which can be conjugated directly to a particle or conjugated to a particle linker that is conjugated to the particle, and the mannose can bind to a mannose receptor on the surface of a macrophage or dendritic cell. In other embodiments, a ligand can be glutamate, which can be conjugated directly to a particle or conjugated to a particle linker that is conjugated to the particle, and the glutamate can bind to a glutamate receptor on the surface of a central nervous system (CNS) cell, e.g. neuron or glial cell, or dendrite of a postsynaptic cell.

In other embodiments, a ligand can be PD-L1 or PD-L2, which can be conjugated directly to a particle or conjugated to a particle linker that is conjugated to the particle, and the PD-L1 or PD-L2 ligand can bind to a PD-1 receptor on the surface of an antigen presenting cell (APC), e.g., a dendritic cell, or a T cell. However, other appropriate ligands may be used for targeting to a particular receptor.

In some embodiments, without limitation, the ligand is a ligand of a PD-1 receptor (e.g., PD-L1 or PD-L2), a CTLA4 receptor, a glutamate receptor, mannose receptor, natural killer group 2D (NKG2D) receptor, chemokine receptor (e.g., CCL19 or CCL21 ligands of CCR7 receptor, or CXCL9 or CXCL10 ligands of CXCR3 receptor), pattern recognition receptor (PRR), Toll-like receptor (TLR), killer activated receptor (KAR), killer inhibitor receptor (KIR), complement receptor, Fc receptor, B cell receptor, or T cell receptor, acetylcholine (ACh) receptor (e.g., nicotine acetylcholine receptor (nAChR) or muscarinic or metabotropic acetylcholine receptor (mAChR)), metabotropic receptor (e.g., G-protein coupled receptor or G-protein coupled receptor linked to a second messenger or second messenger system), ionotropic receptor (e.g., N-methyl D-aspartate (NMDA), amino methyl propionic acid (AMPA), Kainite), transferrin receptor, adenosine receptor (AR), 5-HT receptor (5-HT₁, 5-HT₂, or 5-HT₃), insulin receptor, γ-aminobutyric acid, 4-aminobutanoic acid (GABA) receptor, dopamine receptor (including D1-type and D2-type), or adrenergic receptor (α or β). In some embodiments, the ligand is glutamate, aspartate, mannose, N-acetylglucosamine, fucose, an NKG2D ligand (e.g. MICA or MICB), B7-H1 (PD-L1), B7-DC (PD-L2), CD80, CD86, delta-like ligand 4 (DLL4), transferrin, insulin, Apo-protein, IGF-1, leptin, NMDA, AMPA, GABA, bicuculline, picrotoxinin, dopamine, nicotine, muscarine, acetylcholine, adenosine, serotonin, phenylephrine, adenylate cyclase, or melatonin. In some embodiments, a ligand facilitates blood brain barrier transport (e.g., a transferrin ligand may be used to target a transferrin binding receptor in cells of the brain vasculature to facilitate blood brain barrier transport). In some embodiments, the ligand facilitates blood brain barrier transport via receptor-mediated transcytosis.

In some embodiments, without limitation, the ligand receptor is a PD-1 receptor, CTLA4 receptor, glutamate receptor, mannose receptor, natural killer group 2D (NKG2D) receptor, chemokine receptor, pattern recognition receptor (PRR), Toll-like receptor (TLR), killer activated receptor (KAR), killer inhibitor receptor (KIR), complement receptor, Fc receptor, B cell receptor, or T cell receptor, acetylcholine (ACh) receptor (e.g., nicotine acetylcholine receptor (nAChR) or muscarinic or metabotropic acetylcholine receptor (mAChR)), metabotropic receptor (e.g., G-protein coupled receptor or G-protein coupled receptor linked to a second messenger or second messenger system), ionotropic receptor (e.g., N-methyl D-aspartate (NMDA), amino methyl propionic acid (AMPA), Kainite), transferrin receptor, adenosine receptor (AR), a 5-HT receptor (5-HT₁, 5-HT₂, or 5-HT₃), insulin receptor, γ-aminobutyric acid, 4-aminobutanoic acid (GABA) receptor, dopamine receptor (including D1-type and D2-type), or adrenergic receptor (α or β).

In some embodiments the cell or tissue to be targeted is in vitro. In some embodiments the cell or tissue to be targeted is in vivo. In some embodiments, without limitation, the cell or tissue that has the ligand receptor is a cell or tissue of the immune system or central nervous system (CNS). In some embodiments, the tissue is a tissue of the central nervous system, muscle, or immune system. In some embodiments, the cell is an antigen presenting cell (APC), e.g., dendritic cell, macrophage, or B cell, a T cell, cytotoxic T cell, natural killer (NK) cell, T helper (T_h) cell, neuron, projection neuron, interneuron, glial cell, microglial cell, astrocyte, oligodendrocyte, ependymal cell, radial glial cell, or dendrite. In some embodiments, the cell is from a cell line that was derived from a normal, transformed or immortalized cell, e.g., a normal, transformed or immortalized immune cell or central nervous system cell.

Other cells may be targeted by using appropriate cell targeting ligands. For example, in some embodiments, cells to be targeted are stem cells, e.g., embryonic stem cells, mesenchymal stem cells, hematopoietic stem cells, cancer stem cells, stromal cells etc. In some embodiments, cells to be targeted are epithelial cells (e.g., corneal epithelial cells, mammary epithelial cells, etc.), fibroblasts, myoblasts (e.g., human skeletal myoblasts), keratinocytes, endothelial cells (e.g., vascular endothelial cells), neural cells, smooth muscle cells, marrow cells, bone cells (e.g., osteocytes, osteoblasts, osteoclasts) hematopoietic cells (e.g., monocytes, macrophages, megakaryocytes, etc.) or placental cells.

Oligonucleotides

Oligonucleotides provided herein are useful for stabilizing RNAs by inhibiting or preventing degradation of the RNAs (e.g., degradation mediated by exonucleases). Such oligonucleotides may be referred to as “stabilizing oligonucleotides.” In some embodiments, oligonucleotides hybridize at a 5′ and/or 3′ region of the RNA resulting in duplex regions that stabilize the RNA by preventing degradation by exonucleotides having single strand processing activity.

In some embodiments, oligonucleotides are provided having a region complementary with at least 5 consecutive nucleotides of a 5′ region of an RNA transcript. In some embodiments, oligonucleotides are provided having a region complementary with at least 5 consecutive nucleotides of a 3′-region of an RNA transcript. In some embodiments, oligonucleotides are provided having a first region complementary with at least 5 consecutive nucleotides of a 5′ region of an RNA transcript, and a second region complementary with at least 5 consecutive nucleotides of a 3′-region of an RNA transcript.

In some embodiments, oligonucleotides are provided having a region complementary with at least 5 consecutive nucleotides of the 5′-UTR of an mRNA transcript. In some embodiments, oligonucleotides are provided having a region complementary with at least 5 consecutive nucleotides of the 3′-UTR, poly(A) tail, or overlapping the polyadenylation junction of the mRNA transcript. In some embodiments, oligonucleotides are provided having a first region complementary with at least 5 consecutive nucleotides of the 5′-UTR of an mRNA transcript, and a second region complementary with at least 5 consecutive nucleotides of the 3′-UTR, poly(A) tail, or overlapping the polyadenylation junction of the mRNA transcript.

In some embodiments, oligonucleotides are provided that have a region of complementarity that is complementary to an RNA transcript in proximity to the 5′-end of the RNA transcript. In such embodiments, the nucleotide at the 3′-end of the region of complementarity of the oligonucleotides may be complementary with the RNA transcript at a position that is within 10 nucleotides, within 20 nucleotides, within 30 nucleotides, within 40 nucleotides, within 50 nucleotides, or within 100 nucleotides, within 200 nucleotides, within 300 nucleotides, within 400 nucleotides or more of the transcription start site of the RNA transcript.

In some embodiments, oligonucleotides are provided that have a region of complementarity that is complementary to an RNA transcript in proximity to the 3′-end of the RNA transcript. In such embodiments, the nucleotide at the 3′-end and/or 5′ end of the region of complementarity may be complementary with the RNA transcript at a position that is within 10 nucleotides, within 20 nucleotides, within 30 nucleotides, within 40 nucleotides, within 50 nucleotides, within 100 nucleotides, within 200 nucleotides, within 300 nucleotides, within 400 nucleotides or more of the 3′-end of the RNA transcript. In some embodiments, if the target RNA transcript is polyadenylated, the nucleotide at the 3′-end of the region of complementarity of the oligonucleotide may be complementary with the RNA transcript at a position that is within 10 nucleotides, within 20 nucleotides, within 30 nucleotides, within 40 nucleotides, within 50 nucleotides, within 100 nucleotides, within 200 nucleotides, within 300 nucleotides, within 400 nucleotides or more of polyadenylation junction. In some embodiments, an oligonucleotide that targets a 3′ region of an RNA comprises a region of complementarity that is a stretch of pyrimidines (e.g., 4 to 10 or 5 to 15 thymine nucleotides) complementary with adenines.

In some embodiments, combinations of 5′ targeting and 3′ targeting oligonucleotides are contacted with a target RNA. In some embodiments, the 5′ targeting and 3′ targeting oligonucleotides a linked together via a linker (e.g., a stretch of nucleotides non-complementary with the target RNA). In some embodiments, the region of complementarity of the 5′ targeting oligonucleotide is complementary to a region in the target RNA that is at least 2, 5, 10, 20, 50, 100, 500, 1000, 5000, 10000 nucleotides upstream from the region of the target RNA that is complementary to the region of complementarity of the 3′ end targeting oligonucleotide.

In some embodiments, oligonucleotides are provided that have the general formula 5′-X₁—X₂-3′, in which X₁has a region of complementarity that is complementary with an RNA transcript (e.g., with at least 5 contiguous nucleotides of the RNA transcript). In some embodiments, the nucleotide at the 3′-end of the region of complementary of X₁may be complementary with a nucleotide in proximity to the transcription start site of the RNA transcript. In some embodiments, the nucleotide at the 3′-end of the region of complementary of X₁may be complementary with a nucleotide that is present within 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides of the transcription start site of the RNA transcript. In some embodiments, the nucleotide at the 3′-end of the region of complementary of X₁may be complementary with the nucleotide at the transcription start site of the RNA transcript.

In some embodiments, X₁comprises 5 to 10 nucleotides, 5 to 15 nucleotides, 5 to 25 nucleotides, 10 to 25 nucleotides, 5 to 20 nucleotides, or 15 to 30 nucleotides. In some embodiments, X₁comprises 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or more nucleotides. In some embodiments, the region of complementarity of X₁may be complementary with at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 contiguous nucleotides of the RNA transcript. In some embodiments, the region of complementarity of X₁may be complementary with 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more contiguous nucleotides of the RNA transcript.

In some embodiments, X₂is absent. In some embodiments, X₂comprises 1 to 10, 1 to 20 nucleotides, 1 to 25 nucleotides, 5 to 20 nucleotides, 5 to 30 nucleotides, 5 to 40 nucleotides, or 5 to 50 nucleotides. In some embodiments, X₂comprises 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50 or more nucleotides. In some embodiments, X₂comprises a region of complementarity complementary with at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 contiguous nucleotides of the RNA transcript. In some embodiments, X₂comprises a region of complementarity complementary with 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more contiguous nucleotides of the RNA transcript.

In some embodiments, the RNA transcript has a 7-methylguanosine cap at its 5′-end. In some embodiments, the nucleotide at the 3′-end of the region of complementary of X₁is complementary with the nucleotide of the RNA transcript that is immediately internal to the 7-methylguanosine cap or in proximity to the cap (e.g., with 10 nucleotides of the cap). In some embodiments, at least the first nucleotide at the 5′-end of X₂is a pyrimidine complementary with guanine (e.g., a cytosine or analogue thereof). In some embodiments, the first and second nucleotides at the 5′-end of X₂are pyrimidines complementary with guanine. Thus, in some embodiments, at least one nucleotide at the 5′-end of X₂is a pyrimidine that may form stabilizing hydrogen bonds with the 7-methylguanosine of the cap.

In some embodiments, X₂forms a stem-loop structure. In some embodiments, X₂comprises the formula 5′-Y₁-Y₂—Y₃-3′, in which X₂forms a stem-loop structure having a loop region comprising the nucleotides of Y₂and a stem region comprising at least two contiguous nucleotides of Y₁hybridized with at least two contiguous nucleotides of Y₃. In some embodiments, the stem region comprises 1-6, 1-5, 2-5, 1-4, 2-4 or 2-3 nucleotides. In some embodiments, the stem region comprises LNA nucleotides. In some embodiments, the stem region comprises 1-6, 1-5, 2-5, 1-4, 2-4 or 2-3 LNA nucleotides. In some embodiments, Y₁and Y₃independently comprise 2 to 10 nucleotides, 2 to 20 nucleotides, 2 to 25 nucleotides, or 5 to 20 nucleotides. In some embodiments, Y₁and Y₃independently comprise 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25 or more nucleotides. In some embodiments, Y₂comprises 3 to 10 nucleotides, 3 to 15 nucleotides, 3 to 25 nucleotides, or 5 to 20 nucleotides. In some embodiments, Y₂comprises 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25 or more nucleotides. In some embodiments, Y₂comprises 2-8, 2-7, 2-6, 2-5, 3-8, 3-7, 3-6, 3-5 or 3-4 nucleotides. In some embodiments, Y₂comprises at least one DNA nucleotide. In some embodiments, the nucleotides of Y₂comprise at least one (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25 or more adenines). In some embodiments, Y₃comprises 1-5, 1-4, 1-3 or 1-2 nucleotides following the 3′ end of the stem region. In some embodiments, the nucleotides of Y₃following the 3′ end of the stem region are DNA nucleotides. In some embodiments, Y₃comprises a pyrimidine complementary with guanine (e.g., cytosine or an analogue thereof). In some embodiments, Y₃comprises one or more (e.g., two) pyrimidines complementary with guanine at a position following the 3′-end of the stem region (e.g., 1, 2, 3 or more nucleotide after the 3′-end of the stem region). Thus, in embodiments where the RNA transcript is capped, Y₃may have a pyrimidine that forms stabilizing hydrogen bonds with the 7-methylguanosine of the cap.

In some embodiments, X₁and X₂are complementary with non-overlapping regions of the RNA transcript. In some embodiments, X₁comprises a region complementary with a 5′ region of the RNA transcript and X₂comprises a region complementary with a 3′ region of the RNA transcript. For example, if the RNA transcript is polyadenylated, X₂may comprise a region of complementarity that is complementary with the RNA transcript at a region within 100 nucleotides, within 50 nucleotides, within 25 nucleotides or within 10 nucleotides of the polyadenylation junction of the RNA transcript. In some embodiments, X₂comprises a region of complementarity that is complementary with the RNA transcript immediately adjacent to or overlapping the polyadenylation junction of the RNA transcript. In some embodiments, X₂comprises at least 2 consecutive pyrimidine nucleotides (e.g., 5 to 15 pyrimidine nucleotides) complementary with adenine nucleotides of the poly(A) tail of the RNA transcript.

In some embodiments, oligonucleotides are provided that comprise the general formula 5′-X₁—X₂-3′, in which X₁comprises at least 2 nucleotides that form base pairs with adenine (e.g., thymidines or uridines or analogues thereof); and X₂comprises a region of complementarity that is complementary with at least 3 contiguous nucleotides of a poly-adenylated RNA transcript, wherein the nucleotide at the 5′-end of the region of complementary of X₂is complementary with the nucleotide of the RNA transcript that is immediately internal to the poly-adenylation junction of the RNA transcript. In such embodiments, X₁may comprises 2 to 10, 2 to 20, 5 to 15 or 5 to 25 nucleotides and X₂may independently comprises 2 to 10, 2 to 20, 5 to 15 or 5 to 25 nucleotides.

In some embodiments, compositions are provided that comprise a first oligonucleotide comprising at least 5 nucleotides (e.g., of 5 to 25 nucleotides) linked through internucleoside linkages, and a second oligonucleotide comprising at least 5 nucleotides (e.g., of 5 to 25 nucleotides) linked through internucleoside linkages, in which the first oligonucleotide is complementary with at least 5 consecutive nucleotides in proximity to the 5′-end of an RNA transcript and the second oligonucleotide is complementary with at least 5 consecutive nucleotides in proximity to the 3′-end of an RNA transcript. In some embodiments, the 5′ end of the first oligonucleotide is linked with the 3′ end of the second oligonucleotide. In some embodiments, the 3′ end of the first oligonucleotide is linked with the 5′ end of the second oligonucleotide. In some embodiments, the 5′ end of the first oligonucleotide is linked with the 5′ end of the second oligonucleotide. In some embodiments, the 3′ end of the first oligonucleotide is linked with the 3′ end of the second oligonucleotide.

In some embodiments, the first oligonucleotide and second oligonucleotide are joined by a linker. The term “linker” generally refers to a chemical moiety that is capable of covalently linking two or more oligonucleotides. In some embodiments, a linker is resistant to cleavage in certain biological contexts, such as in a mammalian cell extract, such as an endosomal extract. However, in some embodiments, at least one bond comprised or contained within the linker is capable of being cleaved (e.g., in a biological context, such as in a mammalian extract, such as an endosomal extract), such that at least two oligonucleotides are no longer covalently linked to one another after bond cleavage. In some embodiments, the linker is not an oligonucleotide having a sequence complementary with the RNA transcript. In some embodiments, the linker is an oligonucleotide (e.g., 2-8 thymines). In some embodiments, the linker is a polypeptide. Other appropriate linkers may also be used, including, for example, linkers disclosed in International Patent Application Publication WO 2013/040429 A1, published on Mar. 21, 2013, and entitled MULTIMERIC ANTISENSE OLIGONUCLEOTIDES, and in International Patent Application Publication WO 2014043544 A1, published on Mar. 20, 2014, and entitled MULTIMERIC ANTISENSE OLIGONUCLEOTIDES. The contents of these publications relating to linkers are incorporated herein by reference in their entireties.

An oligonucleotide may have a region of complementarity with a target RNA transcript (e.g., a mammalian mRNA transcript) that has less than a threshold level of complementarity with every sequence of nucleotides, of equivalent length, of an off-target RNA transcript. For example, an oligonucleotide may be designed to ensure that it does not have a sequence that targets RNA transcripts in a cell other than the target RNA transcript. The threshold level of sequence identity may be 50%, 60%, 70%, 80%, 85%, 90%, 95%, 99% or 100% sequence identity.

An oligonucleotide may be complementary to RNA transcripts encoded by homologues of a gene across different species (e.g., a mouse, rat, rabbit, goat, monkey, etc.) In some embodiments, oligonucleotides having these characteristics may be tested in vivo or in vitro for efficacy in multiple species (e.g., human and mouse). This approach also facilitates development of clinical candidates for treating human disease by selecting a species in which an appropriate animal exists for the disease.

In some embodiments, the region of complementarity of an oligonucleotide is complementary with at least 8 to 15, 8 to 30, 8 to 40, or 10 to 50, or 5 to 50, or 5 to 40 bases, e.g., 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 consecutive nucleotides of a target RNA. In some embodiments, the region of complementarity is complementary with at least 8 consecutive nucleotides of a target RNA.

Complementary, as the term is used in the art, refers to the capacity for precise pairing between two nucleotides. For example, if a nucleotide at a certain position of an oligonucleotide is capable of hydrogen bonding with a nucleotide at a corresponding position of a target RNA, then the nucleotide of the oligonucleotide and the nucleotide of the target RNA are complementary to each other at that position. The oligonucleotide and target RNA are complementary to each other when a sufficient number of corresponding positions in each molecule are occupied by nucleotides that can hydrogen bond with each other through their bases. Thus, “complementary” is a term which is used to indicate a sufficient degree of complementarity or precise pairing such that stable and specific binding occurs between the oligonucleotide and target RNA. For example, if a base at one position of an oligonucleotide is capable of hydrogen bonding with a base at the corresponding position of a target RNA, then the bases are considered to be complementary to each other at that position. 100% complementarity is not required.

An oligonucleotide may be at least 80% complementary to (optionally one of at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% complementary to) the consecutive nucleotides of a target RNA. In some embodiments an oligonucleotide may contain 1, 2 or 3 base mismatches compared to the portion of the consecutive nucleotides of the target RNA. In some embodiments an oligonucleotide may have up to 3 mismatches over 15 bases, or up to 2 mismatches over 10 bases.

In some embodiments, a complementary nucleic acid sequence need not be 100% complementary to that of its target to be specifically hybridizable. In some embodiments, an oligonucleotide for purposes of the present disclosure is specifically hybridizable with a target RNA when hybridization of the oligonucleotide to the target RNA prevents or inhibits degradation of the target RNA, and when there is a sufficient degree of complementarity to avoid non-specific binding of the sequence to non-target sequences under conditions in which avoidance of non-specific binding is desired, e.g., under physiological conditions in the case of in vivo assays or therapeutic treatment, and in the case of in vitro assays, under conditions in which the assays are performed under suitable conditions of stringency.

In some embodiments, an oligonucleotide is 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 60, 70, 80 or more nucleotides in length. In some embodiments, the oligonucleotide is 8 to 50, 10 to 30, 9 to 20, 15 to 30 or 8 to 80 nucleotides in length.

Base pairings may include both canonical Watson-Crick base pairing and non-Watson-Crick base pairing (e.g., Wobble base pairing and Hoogsteen base pairing). It is understood that for complementary base pairings, adenosine-type bases (A) are complementary to thymidine-type bases (T) or uracil-type bases (U), that cytosine-type bases (C) are complementary to guanosine-type bases (G), and that universal bases such as 3-nitropyrrole or 5-nitroindole can hybridize to and are considered complementary to any A, C, U, or T. Inosine (I) has also been considered in the art to be a universal base and is considered complementary to any A, C, U or T.

In some embodiments, any one or more thymidine (T) nucleotides (or modified nucleotide thereof) or uridine (U) nucleotides (or a modified nucleotide thereof) in a sequence provided herein, including a sequence provided in the sequence listing, may be replaced with any other nucleotide suitable for base pairing (e.g., via a Watson-Crick base pair) with an adenosine nucleotide. In some embodiments, any one or more thymidine (T) nucleotides (or modified nucleotide thereof) or uridine (U) nucleotides (or a modified nucleotide thereof) in a sequence provided herein, including a sequence provided in the sequence listing, may be suitably replaced with a different pyrimidine nucleotide or vice versa. In some embodiments, any one or more thymidine (T) nucleotides (or modified nucleotide thereof) in a sequence provided herein, including a sequence provided in the sequence listing, may be suitably replaced with a uridine (U) nucleotide (or a modified nucleotide thereof) or vice versa.

In some embodiments, an oligonucleotide may have a sequence that does not contain guanosine nucleotide stretches (e.g., 3 or more, 4 or more, 5 or more, 6 or more consecutive guanosine nucleotides). In some embodiments, oligonucleotides having guanosine nucleotide stretches have increased non-specific binding and/or off-target effects, compared with oligonucleotides that do not have guanosine nucleotide stretches. Contiguous runs of three or more Gs or Cs may not be preferable in some embodiments. Accordingly, in some embodiments, the oligonucleotide does not comprise a stretch of three or more guanosine nucleotides.

An oligonucleotide may have a sequence that is has greater than 30% G-C content, greater than 40% G-C content, greater than 50% G-C content, greater than 60% G-C content, greater than 70% G-C content, or greater than 80% G-C content. An oligonucleotide may have a sequence that has up to 100% G-C content, up to 95% G-C content, up to 90% G-C content, or up to 80% G-C content. In some embodiments, GC content of an oligonucleotide is preferably between about 30-60%.

It is to be understood that any oligonucleotide provided herein can be excluded.

In some embodiments, it has been found that oligonucleotides disclosed herein may increase stability of a target RNA by at least about 50% (e.g. 150% of normal or 1.5 fold), or by about 2 fold to about 5 fold. In some embodiments, stability (e.g., stability in a cell) may be increased by at least about 15 fold, 20 fold, 30 fold, 40 fold, 50 fold or 100 fold, or any range between any of the foregoing numbers. In some embodiments, increased mRNA stability has been shown to correlate to increased protein expression. Similarly, in some embodiments, increased stability of non-coding positively correlates with increased activity of the RNA.

It is understood that any reference to uses of oligonucleotides or other molecules throughout the description contemplates use of the oligonucleotides or other molecules in preparation of a pharmaceutical composition or medicament for use in the treatment of condition or a disease associated with decreased levels or activity of a RNA transcript. Thus, as one nonlimiting example, this aspect of the disclosure includes use of oligonucleotides or other molecules in the preparation of a medicament for use in the treatment of disease, wherein the treatment involves posttranscriptionally altering protein and/or RNA levels in a targeted manner.

Methods for identifying transcript start sites and polyadenylation junctions are known in the art and may be used in selecting oligonucleotides that specifically bind to these regions for stabilizing RNA transcripts. In some embodiments, 3′ end oligonucleotides may be designed by identifying RNA 3′ ends using quantitative end analysis of poly-A tails. In some embodiments, 5′ end oligonucleotides may be designed by identifying 5′ start sites using Cap analysis gene expression (CAGE). Appropriate methods are disclosed, for example, in Ozsolak et al. Comprehensive Polyadenylation Site Maps in Yeast and Human Reveal Pervasive Alternative Polyadenylation. Cell. Volume 143, Issue 6, 2010, Pages 1018-1029; Shiraki, T, et al., Cap analysis gene expression for high-throughput analysis of transcriptional starting point and identification of promoter usage. Proc Natl Acad Sci USA. 100 (26): 15776-81. 2003 Dec. 23; and Zhao, X, et al., (2011). Systematic Clustering of Transcription Start Site Landscapes. PLoS ONE (Public Library of Science) 6 (8): e23409, the contents of each of which are incorporated herein by reference. Other appropriate methods for identifying transcript start sites and polyadenylation junctions may also be used, including, for example, RNA-Paired-end tags (PET) (See, e.g., Ruan X, Ruan Y. Methods Mol Biol. 2012; 809:535-62); use of standard EST databases; RACE combined with microarray or sequencing, PAS-Seq (See, e.g., Peter J. Shepard, et al., RNA. 2011 April; 17(4): 761-772); and 3P-Seq (See, e.g., Calvin H. Jan, Nature. 2011 Jan. 6; 469(7328): 97-101; and others.

Exemplary oligonucleotides targeting the 5′ and/or 3′ ends of RNAs are provided in Example 2 and Tables 3-13.

Oligonucleotide Modifications

In some embodiments, oligonucleotides are provided with chemistries suitable for delivery, hybridization and stability within cells to target and stabilize RNA transcripts. Furthermore, in some embodiments, oligonucleotide chemistries are provided that are useful for controlling the pharmacokinetics, biodistribution, bioavailability and/or efficacy of the oligonucleotides. Accordingly, oligonucleotides described herein may be modified, e.g., comprise a modified sugar moiety, a modified internucleoside linkage, a modified nucleotide and/or combinations thereof. In addition, the oligonucleotides may exhibit one or more of the following properties: do not induce substantial cleavage or degradation of the target RNA; do not cause substantially complete cleavage or degradation of the target RNA; do not activate the RNAse H pathway; do not activate RISC; do not recruit any Argonaute family protein; are not cleaved by Dicer; do not mediate alternative splicing; are not immune stimulatory; are nuclease resistant; have improved cell uptake compared to unmodified oligonucleotides; are not toxic to cells or mammals; and may have improved endosomal exit.

Oligonucleotides that are designed to interact with RNA to modulate gene expression are a distinct subset of base sequences from those that are designed to bind a DNA target (e.g., are complementary to the underlying genomic DNA sequence from which the RNA is transcribed).

Any of the oligonucleotides disclosed herein may be linked to one or more other oligonucleotides disclosed herein by a linker, e.g., a cleavable linker.

Oligonucleotides of the disclosure can be stabilized against nucleolytic degradation such as by the incorporation of a modification, e.g., a nucleotide modification. For example, nucleic acid sequences of the disclosure include a phosphorothioate at least the first, second, or third internucleotide linkage at the 5′ or 3′ end of the nucleotide sequence. As another example, the nucleic acid sequence can include a 2′-modified nucleotide, e.g., a 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O—NMA). As another example, the nucleic acid sequence can include at least one 2′-O-methyl-modified nucleotide, and in some embodiments, all of the nucleotides include a 2′-O-methyl modification. In some embodiments, the nucleic acids are “locked,” e.g., comprise nucleic acid analogues in which the ribose ring is “locked” by a methylene bridge connecting the 2′-O atom and the 4′-C atom.

Any of the modified chemistries or formats of oligonucleotides described herein can be combined with each other, and that one, two, three, four, five, or more different types of modifications can be included within the same molecule.

In some embodiments, the oligonucleotide may comprise at least one ribonucleotide, at least one deoxyribonucleotide, and/or at least one bridged nucleotide. In some embodiments, the oligonucleotide may comprise a bridged nucleotide, such as a locked nucleic acid (LNA) nucleotide, a constrained ethyl (cEt) nucleotide, or an ethylene bridged nucleic acid (ENA) nucleotide. Examples of such nucleotides are disclosed herein and known in the art. In some embodiments, the oligonucleotide comprises a nucleotide analog disclosed in one of the following United States patent or patent application Publications: U.S. Pat. No. 7,399,845, U.S. Pat. No. 7,741,457, U.S. Pat. No. 8,022,193, U.S. Pat. No. 7,569,686, U.S. Pat. No. 7,335,765, U.S. Pat. No. 7,314,923, U.S. Pat. No. 7,335,765, and U.S. Pat. No. 7,816,333, US 20110009471, the entire contents of each of which are incorporated herein by reference for all purposes. The oligonucleotide may have one or more 2′ O-methyl nucleotides. The oligonucleotide may consist entirely of 2′ O-methyl nucleotides.

Often an oligonucleotide has one or more nucleotide analogues. For example, an oligonucleotide may have at least one nucleotide analogue that results in an increase in T_mof the oligonucleotide in a range of 1° C., 2° C., 3° C., 4° C., or 5° C. compared with an oligonucleotide that does not have the at least one nucleotide analogue. An oligonucleotide may have a plurality of nucleotide analogues that results in a total increase in T_mof the oligonucleotide in a range of 2° C., 3° C., 4° C., 5° C., 6° C., 7° C., 8° C., 9° C., 10° C., 15° C., 20° C., 25° C., 30° C., 35° C., 40° C., 45° C. or more compared with an oligonucleotide that does not have the nucleotide analogue.

The oligonucleotide may be of up to 50 nucleotides in length in which 2 to 10, 2 to 15, 2 to 16, 2 to 17, 2 to 18, 2 to 19, 2 to 20, 2 to 25, 2 to 30, 2 to 40, 2 to 45, or more nucleotides of the oligonucleotide are nucleotide analogues. The oligonucleotide may be of 8 to 30 nucleotides in length in which 2 to 10, 2 to 15, 2 to 16, 2 to 17, 2 to 18, 2 to 19, 2 to 20, 2 to 25, 2 to 30 nucleotides of the oligonucleotide are nucleotide analogues. The oligonucleotide may be of 8 to 15 nucleotides in length in which 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 2 to 10, 2 to 11, 2 to 12, 2 to 13, 2 to 14 nucleotides of the oligonucleotide are nucleotide analogues. Optionally, the oligonucleotides may have every nucleotide except 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides modified.

The oligonucleotide may consist entirely of bridged nucleotides (e.g., LNA nucleotides, cEt nucleotides, ENA nucleotides). The oligonucleotide may comprise alternating deoxyribonucleotides and 2′-fluoro-deoxyribonucleotides. The oligonucleotide may comprise alternating deoxyribonucleotides and 2′-O-methyl nucleotides. The oligonucleotide may comprise alternating deoxyribonucleotides and ENA nucleotide analogues. The oligonucleotide may comprise alternating deoxyribonucleotides and LNA nucleotides. The oligonucleotide may comprise alternating LNA nucleotides and 2′-O-methyl nucleotides. The oligonucleotide may have a 5′ nucleotide that is a bridged nucleotide (e.g., a LNA nucleotide, cEt nucleotide, ENA nucleotide). The oligonucleotide may have a 5′ nucleotide that is a deoxyribonucleotide.

The oligonucleotide may comprise deoxyribonucleotides flanked by at least one bridged nucleotide (e.g., a LNA nucleotide, cEt nucleotide, ENA nucleotide) on each of the 5′ and 3′ ends of the deoxyribonucleotides. The oligonucleotide may comprise deoxyribonucleotides flanked by 1, 2, 3, 4, 5, 6, 7, 8 or more bridged nucleotides (e.g., LNA nucleotides, cEt nucleotides, ENA nucleotides) on each of the 5′ and 3′ ends of the deoxyribonucleotides. The 3′ position of the oligonucleotide may have a 3′ hydroxyl group. The 3′ position of the oligonucleotide may have a 3′ thiophosphate.

The oligonucleotide may be conjugated with a label. For example, the oligonucleotide may be conjugated with a biotin moiety, cholesterol, Vitamin A, folate, sigma receptor ligands, aptamers, peptides, such as CPP, hydrophobic molecules, such as lipids, ligands of the asialoglycoprotein receptor (ASGPR), such as GalNac, or dynamic polyconjugates and variants thereof at its 5′ or 3′ end.

Preferably an oligonucleotide comprises one or more modifications comprising: a modified sugar moiety, and/or a modified internucleoside linkage, and/or a modified nucleotide and/or combinations thereof. It is not necessary for all positions in a given oligonucleotide to be uniformly modified, and in fact more than one of the modifications described herein may be incorporated in a single oligonucleotide or even at within a single nucleoside within an oligonucleotide.

In some embodiments, the oligonucleotides are chimeric oligonucleotides that contain two or more chemically distinct regions, each made up of at least one nucleotide. These oligonucleotides typically contain at least one region of modified nucleotides that confers one or more beneficial properties (such as, for example, increased nuclease resistance, increased uptake into cells, increased binding affinity for the target) and a region that is a substrate for enzymes capable of cleaving RNA:DNA or RNA:RNA hybrids. Chimeric oligonucleotides of the disclosure may be formed as composite structures of two or more oligonucleotides, modified oligonucleotides, oligonucleosides and/or oligonucleotide mimetics as described above. Such compounds have also been referred to in the art as hybrids or gapmers. Representative United States patents that teach the preparation of such hybrid structures comprise, but are not limited to, U.S. Pat. Nos. 5,013,830; 5,149,797; 5,220,007; 5,256,775; 5,366,878; 5,403,711; 5,491,133; 5,565,350; 5,623,065; 5,652,355; 5,652,356; and 5,700,922, each of which is herein incorporated by reference.

In some embodiments, an oligonucleotide comprises at least one nucleotide modified at the 2′ position of the sugar, most preferably a 2′-O-alkyl, 2′-O-alkyl-O-alkyl or 2′-fluoro-modified nucleotide. In other preferred embodiments, RNA modifications include 2′-fluoro, 2′-amino and 2′ O-methyl modifications on the ribose of pyrimidines, abasic residues or an inverted base at the 3′ end of the RNA. Such modifications are routinely incorporated into oligonucleotides and these oligonucleotides have been shown to have a higher Tm (e.g., higher target binding affinity) than 2′-deoxyoligonucleotides against a given target.

A number of nucleotide and nucleoside modifications have been shown to make the oligonucleotide into which they are incorporated more resistant to nuclease digestion than the native oligodeoxynucleotide; these modified oligos survive intact for a longer time than unmodified oligonucleotides. Specific examples of modified oligonucleotides include those comprising modified backbones, for example, phosphorothioates, phosphotriesters, methyl phosphonates, short chain alkyl or cycloalkyl intersugar linkages or short chain heteroatomic or heterocyclic intersugar linkages. In some embodiments, oligonucleotides may have phosphorothioate backbones; heteroatom backbones, such as methylene(methylimino) or MMI backbones; amide backbones (see De Mesmaeker et al. Ace. Chem. Res. 1995, 28:366-374); morpholino backbones (see Summerton and Weller, U.S. Pat. No. 5,034,506); or peptide nucleic acid (PNA) backbones (wherein the phosphodiester backbone of the oligonucleotide is replaced with a polyamide backbone, the nucleotides being bound directly or indirectly to the aza nitrogen atoms of the polyamide backbone, see Nielsen et al., Science 1991, 254, 1497). Phosphorus-containing linkages include, but are not limited to, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates comprising 3′alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates comprising 3′-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates having normal 3′-5′ linkages, 2′-5′ linked analogs of these, and those having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3′-5′ to 5′-3′ or 2′-5′ to 5′-2′; see U.S. Pat. Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243; 5,177,196; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717; 5,321,131; 5,399,676; 5,405,939; 5,453,496; 5,455, 233; 5,466,677; 5,476,925; 5,519,126; 5,536,821; 5,541,306; 5,550,111; 5,563,253; 5,571,799; 5,587,361; and 5,625,050.

Morpholino-based oligomeric compounds are described in Dwaine A. Braasch and David R. Corey, Biochemistry, 2002, 41(14), 4503-4510); Genesis, volume 30, issue 3, 2001; Heasman, J., Dev. Biol., 2002, 243, 209-214; Nasevicius et al., Nat. Genet., 2000, 26, 216-220; Lacerra et al., Proc. Natl. Acad. Sci., 2000, 97, 9591-9596; and U.S. Pat. No. 5,034,506, issued Jul. 23, 1991. In some embodiments, the morpholino-based oligomeric compound is a phosphorodiamidate morpholino oligomer (PMO) (e.g., as described in Iverson, Curr. Opin. Mol. Ther., 3:235-238, 2001; and Wang et al., J. Gene Med., 12:354-364, 2010; the disclosures of which are incorporated herein by reference in their entireties).

Cyclohexenyl nucleic acid oligonucleotide mimetics are described in Wang et al., J. Am. Chem. Soc., 2000, 122, 8595-8602.

Modified oligonucleotide backbones that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatom and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages. These comprise those having morpholino linkages (formed in part from the sugar portion of a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones; methylene formacetyl and thioformacetyl backbones; alkene containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed N, O, S and CH2 component parts; see U.S. Pat. Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033; 5,264, 562; 5, 264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; 5,489,677; 5,541,307; 5,561,225; 5,596,086; 5,602,240; 5,610,289; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and 5,677,439, each of which is herein incorporated by reference.

Modified oligonucleotides are also known that include oligonucleotides that are based on or constructed from arabinonucleotide or modified arabinonucleotide residues. Arabinonucleosides are stereoisomers of ribonucleosides, differing only in the configuration at the 2′-position of the sugar ring. In some embodiments, a 2′-arabino modification is 2′-F arabino. In some embodiments, the modified oligonucleotide is 2′-fluoro-D-arabinonucleic acid (FANA) (as described in, for example, Lon et al., Biochem., 41:3457-3467, 2002 and Min et al., Bioorg. Med. Chem. Lett., 12:2651-2654, 2002; the disclosures of which are incorporated herein by reference in their entireties). Similar modifications can also be made at other positions on the sugar, particularly the 3′ position of the sugar on a 3′ terminal nucleoside or in 2′-5′ linked oligonucleotides and the 5′ position of 5′ terminal nucleotide.

PCT Publication No. WO 99/67378 discloses arabinonucleic acids (ANA) oligomers and their analogues for improved sequence specific inhibition of gene expression via association to complementary messenger RNA.

Other preferred modifications include ethylene-bridged nucleic acids (ENAs) (e.g., International Patent Publication No. WO 2005/042777, Morita et al., Nucleic Acid Res., Suppl 1:241-242, 2001; Surono et al., Hum. Gene Ther., 15:749-757, 2004; Koizumi, Curr. Opin. Mol. Ther., 8:144-149, 2006 and Horie et al., Nucleic Acids Symp. Ser (Oxf), 49:171-172, 2005; the disclosures of which are incorporated herein by reference in their entireties). Preferred ENAs include, but are not limited to, 2′-O,4′-C-ethylene-bridged nucleic acids.

Examples of LNAs are described in WO/2008/043753 and include compounds of the following general formula.

embedded image

where X and Y are independently selected among the groups —O—,

—S—, —N(H)—, N(R)—, —CH₂— or —CH— (if part of a double bond),

—CH₂—O—, —CH₂—S—, —CH₂—N(H)—, —CH₂—N(R)—, —CH₂—CH₂— or —CH₂—CH— (if part of a double bond),

—CH═CH—, where R is selected from hydrogen and C_1-4-alkyl; Z and Z* are independently selected among an internucleoside linkage, a terminal group or a protecting group; B constitutes a natural or non-natural nucleotide base moiety; and the asymmetric groups may be found in either orientation.

Preferably, the LNA used in the oligonucleotides described herein comprises at least one LNA unit according any of the formulas

embedded image

wherein Y is —O—, —S—, —NH—, or N(R^H); Z and Z* are independently selected among an internucleoside linkage, a terminal group or a protecting group; B constitutes a natural or non-natural nucleotide base moiety, and RH is selected from hydrogen and C_1-4-alkyl.

In some embodiments, the Locked Nucleic Acid (LNA) used in the oligonucleotides described herein comprises at least one Locked Nucleic Acid (LNA) unit according any of the formulas shown in Scheme 2 of PCT/DK2006/000512.

In some embodiments, the LNA used in the oligomer of the disclosure comprises internucleoside linkages selected from -0-P(O)₂—O—, —O—P(O,S)—O—, —O—P(S)₂—O—, —S—P(O)₂—O—, —S—P(O,S)—O—, —S—P(S)₂—O—, —0-P(O)₂—S—, —O—P(O,S)—S—, —S—P(O)₂—S—, —O—PO(R^H)—O—, O—PO(OCH₃)—O—, —O—PO(NR^H)—O—, —0-PO(OCH₂CH₂S—R)—O—, —O—PO(BH₃)—O—, —O—PO(NHR^H)—O—, —O—P(O)₂—NR^H—, —NR^H—P(O)₂—O—, —NR^H—CO—O—, where R^His selected from hydrogen and C_1-4-alkyl.

Other examples of LNA units are shown below:

embedded image

The term “thio-LNA” comprises a locked nucleotide in which at least one of X or Y in the general formula above is selected from S or —CH₂—S—. Thio-LNA can be in both beta-D and alpha-L-configuration.

The term “amino-LNA” comprises a locked nucleotide in which at least one of X or Y in the general formula above is selected from —N(H)—, N(R)—, CH₂—N(H)—, and —CH₂—N(R)— where R is selected from hydrogen and C_1-4-alkyl. Amino-LNA can be in both beta-D and alpha-L-configuration.

The term “oxy-LNA” comprises a locked nucleotide in which at least one of X or Y in the general formula above represents —O— or —CH₂—O—. Oxy-LNA can be in both beta-D and alpha-L-configuration.

The term “ena-LNA” comprises a locked nucleotide in which Y in the general formula above is —CH₂—O— (where the oxygen atom of —CH₂—O— is attached to the 2′-position relative to the base B).

LNAs are described in additional detail herein.

One or more substituted sugar moieties can also be included, e.g., one of the following at the 2′ position: OH, SH, SCH₃, F, OCN, OCH₃OCH₃, OCH₃O(CH₂)n CH₃, O(CH₂)n NH₂or O(CH₂)n CH₃where n is from 1 to about 10; C1 to C10 lower alkyl, alkoxyalkoxy, substituted lower alkyl, alkaryl or aralkyl; C1; Br; CN; CF₃; OCF₃; O-, S-, or N-alkyl; O—, S—, or N-alkenyl; SOCH₃; SO₂CH₃; ONO₂; NO₂; N₃; NH2; heterocycloalkyl; heterocycloalkaryl; aminoalkylamino; polyalkylamino; substituted silyl; an RNA cleaving group; a reporter group; an intercalator; a group for improving the pharmacokinetic properties of an oligonucleotide; or a group for improving the pharmacodynamic properties of an oligonucleotide and other substituents having similar properties. A preferred modification includes 2′-methoxyethoxy [2′-O—CH₂CH2OCH₃, also known as 2′-O-(2-methoxyethyl)](Martin et al, HeIv. Chim. Acta, 1995, 78, 486). Other preferred modifications include 2′-methoxy (2′-O—CH₃), 2′-propoxy (2′-OCH₂CH₂CH₃) and 2′-fluoro (2′-F). Similar modifications may also be made at other positions on the oligonucleotide, particularly the 3′ position of the sugar on the 3′ terminal nucleotide and the 5′ position of 5′ terminal nucleotide. Oligonucleotides may also have sugar mimetics such as cyclobutyls in place of the pentofuranosyl group.

Oligonucleotides can also include, additionally or alternatively, nucleobase (often referred to in the art simply as “base”) modifications or substitutions. As used herein, “unmodified” or “natural” nucleobases include adenine (A), guanine (G), thymine (T), cytosine (C) and uracil (U). Modified nucleobases include nucleobases found only infrequently or transiently in natural nucleic acids, e.g., hypoxanthine, 6-methyladenine, 5-Me pyrimidines, particularly 5-methylcytosine (also referred to as 5-methyl-2′ deoxycytosine and often referred to in the art as 5-Me-C), 5-hydroxymethylcytosine (HMC), glycosyl HMC and gentobiosyl HMC, isocytosine, pseudoisocytosine, as well as synthetic nucleobases, e.g., 2-aminoadenine, 2-(methylamino)adenine, 2-(imidazolylalkyl)adenine, 2-(aminoalklyamino)adenine or other heterosubstituted alkyladenines, 2-thiouracil, 2-thiothymine, 5-bromouracil, 5-hydroxymethyluracil, 5-propynyluracil, 8-azaguanine, 7-deazaguanine, N6 (6-aminohexyl)adenine, 6-aminopurine, 2-aminopurine, 2-chloro-6-aminopurine and 2,6-diaminopurine or other diaminopurines. See, e.g., Kornberg, “DNA Replication,” W. H. Freeman & Co., San Francisco, 1980, pp 75-77; and Gebeyehu, G., et al. Nucl. Acids Res., 15:4513 (1987)). A “universal” base known in the art, e.g., inosine, can also be included. 5-Me-C substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2° C. (Sanghvi, in Crooke, and Lebleu, eds., Antisense Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and may be used as base substitutions.

It is not necessary for all positions in a given oligonucleotide to be uniformly modified, and in fact more than one of the modifications described herein may be incorporated in a single oligonucleotide or even at within a single nucleoside within an oligonucleotide.

In some embodiments, both a sugar and an internucleoside linkage, e.g., the backbone, of the nucleotide units are replaced with novel groups. The base units are maintained for hybridization with an appropriate nucleic acid target compound. One such oligomeric compound, an oligonucleotide mimetic that has been shown to have excellent hybridization properties, is referred to as a peptide nucleic acid (PNA). In PNA compounds, the sugar-backbone of an oligonucleotide is replaced with an amide containing backbone, for example, an aminoethylglycine backbone. The nucleobases are retained and are bound directly or indirectly to aza nitrogen atoms of the amide portion of the backbone. Representative United States patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, each of which is herein incorporated by reference. Further teaching of PNA compounds can be found in Nielsen et al, Science, 1991, 254, 1497-1500.

Oligonucleotides can also include one or more nucleobase (often referred to in the art simply as “base”) modifications or substitutions. As used herein, “unmodified” or “natural” nucleobases comprise the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U). Modified nucleobases comprise other synthetic and natural nucleobases such as 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudo-uracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylquanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine.

Further, nucleobases comprise those disclosed in U.S. Pat. No. 3,687,808, those disclosed in “The Concise Encyclopedia of Polymer Science And Engineering”, pages 858-859, Kroschwitz, ed. John Wiley & Sons, 1990, those disclosed by Englisch et al., Angewandle Chemie, International Edition, 1991, 30, page 613, and those disclosed by Sanghvi, Chapter 15, Antisense Research and Applications,” pages 289-302, Crooke, and Lebleu, eds., CRC Press, 1993. Certain of these nucleobases are particularly useful for increasing the binding affinity of the oligomeric compounds of the disclosure. These include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and 0-6 substituted purines, comprising 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine. 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2<0>C (Sanghvi, et al., eds, “Antisense Research and Applications,” CRC Press, Boca Raton, 1993, pp. 276-278) and are presently preferred base substitutions, even more particularly when combined with 2′-O-methoxyethyl sugar modifications. Modified nucleobases are described in U.S. Pat. No. 3,687,808, as well as U.S. Pat. Nos. 4,845,205; 5,130,302; 5,134,066; 5,175, 273; 5, 367,066; 5,432,272; 5,457,187; 5,459,255; 5,484,908; 5,502,177; 5,525,711; 5,552,540; 5,587,469; 5,596,091; 5,614,617; 5,750,692, and 5,681,941, each of which is herein incorporated by reference.

In some embodiments, the oligonucleotides are chemically linked to one or more moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the oligonucleotide. For example, one or more oligonucleotides, of the same or different types, can be conjugated to each other; or oligonucleotides can be conjugated to targeting moieties with enhanced specificity for a cell type or tissue type. Such moieties include, but are not limited to, lipid moieties such as a cholesterol moiety (Letsinger et al., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid (Manoharan et al., Bioorg. Med. Chem. Let., 1994, 4, 1053-1060), a thioether, e.g., hexyl-S-tritylthiol (Manoharan et al, Ann. N. Y. Acad. Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med. Chem. Let., 1993, 3, 2765-2770), a thiocholesterol (Oberhauser et al., Nucl. Acids Res., 1992, 20, 533-538), an aliphatic chain, e.g., dodecandiol or undecyl residues (Kabanov et al., FEBS Lett., 1990, 259, 327-330; Svinarchuk et al., Biochimie, 1993, 75, 49-54), a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al., Tetrahedron Lett., 1995, 36, 3651-3654; Shea et al., Nucl. Acids Res., 1990, 18, 3777-3783), a polyamine or a polyethylene glycol chain (Mancharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), or adamantane acetic acid (Manoharan et al., Tetrahedron Lett., 1995, 36, 3651-3654), a palmityl moiety (Mishra et al., Biochim. Biophys. Acta, 1995, 1264, 229-237), or an octadecylamine or hexylamino-carbonyl-t oxycholesterol moiety (Crooke et al., J. Pharmacol. Exp. Ther., 1996, 277, 923-937). See also U.S. Pat. Nos. 4,828,979; 4,948,882; 5,218,105; 5,525,465; 5,541,313; 5,545,730; 5,552, 538; 5,578,717, 5,580,731; 5,580,731; 5,591,584; 5,109,124; 5,118,802; 5,138,045; 5,414,077; 5,486, 603; 5,512,439; 5,578,718; 5,608,046; 4,587,044; 4,605,735; 4,667,025; 4,762, 779; 4,789,737; 4,824,941; 4,835,263; 4,876,335; 4,904,582; 4,958,013; 5,082, 830; 5,112,963; 5,214,136; 5,082,830; 5,112,963; 5,214,136; 5, 245,022; 5,254,469; 5,258,506; 5,262,536; 5,272,250; 5,292,873; 5,317,098; 5,371,241, 5,391, 723; 5,416,203, 5,451,463; 5,510,475; 5,512,667; 5,514,785; 5, 565,552; 5,567,810; 5,574,142; 5,585,481; 5,587,371; 5,595,726; 5,597,696; 5,599,923; 5,599, 928 and 5,688,941, each of which is herein incorporated by reference.

These moieties or conjugates can include conjugate groups covalently bound to functional groups such as primary or secondary hydroxyl groups. Conjugate groups of the disclosure include intercalators, reporter molecules, polyamines, polyamides, polyethylene glycols, polyethers, groups that enhance the pharmacodynamic properties of oligomers, and groups that enhance the pharmacokinetic properties of oligomers. Typical conjugate groups include cholesterols, lipids, phospholipids, biotin, phenazine, folate, phenanthridine, anthraquinone, acridine, fluoresceins, rhodamines, coumarins, and dyes. Groups that enhance the pharmacodynamic properties, in the context of this disclosure, include groups that improve uptake, enhance resistance to degradation, and/or strengthen sequence-specific hybridization with the target nucleic acid. Groups that enhance the pharmacokinetic properties, in the context of this disclosure, include groups that improve uptake, distribution, metabolism or excretion of the compounds of the present disclosure. Representative conjugate groups are disclosed in International Patent Application No. PCT/US92/09196, filed Oct. 23, 1992, and U.S. Pat. No. 6,287,860, which are incorporated herein by reference. Conjugate moieties include, but are not limited to, lipid moieties such as a cholesterol moiety, cholic acid, a thioether, e.g., hexyl-5-tritylthiol, a thiocholesterol, an aliphatic chain, e.g., dodecandiol or undecyl residues, a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate, a polyamine or a polyethylene glycol chain, or adamantane acetic acid, a palmityl moiety, or an octadecylamine or hexylamino-carbonyl-oxy cholesterol moiety. See, e.g., U.S. Pat. Nos. 4,828,979; 4,948,882; 5,218,105; 5,525,465; 5,541,313; 5,545,730; 5,552,538; 5,578,717, 5,580,731; 5,580,731; 5,591,584; 5,109,124; 5,118,802; 5,138,045; 5,414,077; 5,486,603; 5,512,439; 5,578,718; 5,608,046; 4,587,044; 4,605,735; 4,667,025; 4,762,779; 4,789,737; 4,824,941; 4,835,263; 4,876,335; 4,904,582; 4,958,013; 5,082,830; 5,112,963; 5,214,136; 5,082,830; 5,112,963; 5,214,136; 5,245,022; 5,254,469; 5,258,506; 5,262,536; 5,272,250; 5,292,873; 5,317,098; 5,371,241, 5,391,723; 5,416,203, 5,451,463; 5,510,475; 5,512,667; 5,514,785; 5,565,552; 5,567,810; 5,574,142; 5,585,481; 5,587,371; 5,595,726; 5,597,696; 5,599,923; 5,599,928 and 5,688,941.

In some embodiments, oligonucleotide modification include modification of the 5′ or 3′ end of the oligonucleotide. In some embodiments, the 3′ end of the oligonucleotide comprises a hydroxyl group or a thiophosphate. It should be appreciated that additional molecules (e.g. a biotin moiety or a fluorophor) can be conjugated to the 5′ or 3′ end of an oligonucleotide. In some embodiments, an oligonucleotide comprises a biotin moiety conjugated to the 5′ nucleotide.

In some embodiments, an oligonucleotide comprises locked nucleic acids (LNA), ENA modified nucleotides, 2′-O-methyl nucleotides, or 2′-fluoro-deoxyribonucleotides. In some embodiments, an oligonucleotide comprises alternating deoxyribonucleotides and 2′-fluoro-deoxyribonucleotides. In some embodiments, an oligonucleotide comprises alternating deoxyribonucleotides and 2′-O-methyl nucleotides. In some embodiments, an oligonucleotide comprises alternating deoxyribonucleotides and ENA modified nucleotides. In some embodiments, an oligonucleotide comprises alternating deoxyribonucleotides and locked nucleic acid nucleotides. In some embodiments, an oligonucleotide comprises alternating locked nucleic acid nucleotides and 2′-O-methyl nucleotides.

In some embodiments, the 5′ nucleotide of the oligonucleotide is a deoxyribonucleotide. In some embodiments, the 5′ nucleotide of the oligonucleotide is a locked nucleic acid nucleotide. In some embodiments, the nucleotides of the oligonucleotide comprise deoxyribonucleotides flanked by at least one locked nucleic acid nucleotide on each of the 5′ and 3′ ends of the deoxyribonucleotides. In some embodiments, the nucleotide at the 3′ position of the oligonucleotide has a 3′ hydroxyl group or a 3′ thiophosphate.

In some embodiments, an oligonucleotide comprises phosphorothioate internucleotide linkages. In some embodiments, an oligonucleotide comprises phosphorothioate internucleotide linkages between at least two nucleotides. In some embodiments, an oligonucleotide comprises phosphorothioate internucleotide linkages between all nucleotides.

It should be appreciated that an oligonucleotide can have any combination of modifications as described herein.

The oligonucleotide may comprise a nucleotide sequence having one or more of the following modification patterns.

(a) (X)Xxxxxx, (X)xXxxxx, (X)xxXxxx, (X)xxxXxx, (X)xxxxXx and (X)xxxxxX,

(b) (X)XXxxxx, (X)XxXxxx, (X)XxxXxx, (X)XxxxXx, (X)XxxxxX, (X)xXXxxx, (X)xXxXxx, (X)xXxxXx, (X)xXxxxX, (X)xxXXxx, (X)xxXxXx, (X)xxXxxX, (X)xxxXXx, (X)xxxXxX and (X)xxxxXX,

(c) (X)XXXxxx, (X)xXXXxx, (X)xxXXXx, (X)xxxXXX, (X)XXxXxx, (X)XXxxXx, (X)XXxxxX, (X)xXXxXx, (X)xXXxxX, (X)xxXXxX, (X)XxXXxx, (X)XxxXXx (X)XxxxXX, (X)xXxXXx, (X)xXxxXX, (X)xxXxXX, (X)xXxXxX and (X)XxXxXx,

(d) (X)xxXXX, (X)xXxXXX, (X)xXXxXX, (X)xXXXxX, (X)xXXXXx, (X)XxxXXXX, (X)XxXxXX, (X)XxXXxX, (X)XxXXx, (X)XXxxXX, (X)XXxXxX, (X)XXxXXx, (X)XXXxxX, (X)XXXxXx, and (X)XXXXxx,

(e) (X)xXXXXX, (X)XxXXXX, (X)XXxXXX, (X)XXXxXX, (X)XXXXxX and (X)XXXXXx, and

(f) XXXXXX, XxXXXXX, XXxXXXX, XXXxXXX, XXXXxXX, XXXXXxX and XXXXXXx, in which “X” denotes a nucleotide analogue, (X) denotes an optional nucleotide analogue, and “x” denotes a DNA or RNA nucleotide unit. Each of the above listed patterns may appear one or more times within an oligonucleotide, alone or in combination with any of the other disclosed modification patterns.

Exemplary oligonucleotide modifications are also provided in Example 2 and Table 14.

RNA and RNA Transcripts

In some embodiments, an RNA transcript targeted by an oligonucleotide disclosed herein is an RNA transcript of a eukaryotic cell. In some embodiments, an RNA transcript targeted by an oligonucleotide disclosed herein is an RNA transcript of a cell of a vertebrate. In some embodiments, an RNA transcript targeted by an oligonucleotide disclosed herein is an RNA transcript of a cell of a mammal, e.g., a primate cell, mouse cell, rat cell, or human cell. In some embodiments, an RNA transcript targeted by an oligonucleotide disclosed herein is an RNA transcript of a cardiomyocyte. In some embodiments, an RNA transcript targeted by an oligonucleotide disclosed herein is an RNA transcribed in the nucleus of a cell. In some embodiments, an RNA transcript targeted by an oligonucleotide disclosed herein is an RNA transcribed in a mitochondrion of a cell. In some embodiments, an RNA transcript targeted by an oligonucleotide disclosed herein is an RNA transcript transcribed by a RNA polymerase II enzyme.

In some embodiments, an RNA transcript targeted by an oligonucleotide disclosed herein is an mRNA expressed from a gene, including, but not limited to, ABCA1, APOA1, ATP2A2, BDNF, FXN, HBA2, HBB, HBD, HBE1, HBG1, HBG2, SMN, UTRN, PTEN, MECP2, and FOXP3, ABCA4, ABCB11, ABCB4, ABCG5, ABCG8, ADIPOQ, ALB, APOE, BCL2L11, BRCA1, CD274, CEP290, CFTR, EPO, F7, F8, FLI1, FMR1, FNDC5, GCH1, GCK, GLP1R, GRN, HAMP, HPRT1, IDO1, IGF1, IL10, IL6, KCNMA1, KCNMB1, KCNMB2, KCNMB3, KCNMB4, KLF1, KLF4, LDLR, MSX2, MYBPC3, NANOG, NF1, NKX2-1, NKX2-1-AS 1, PAH, PTGS2, RB1, RPS 14, RPS 19, SCARB 1, SERPINF1, SIRT1, SIRT6, SMAD7, ST7, STAT3, TSIX, and XIST. RNA transcripts for these and other genes may be selected or identified experimentally, for example, using RNA sequencing (RNA-Seq) or other appropriate methods. RNA transcripts may also be selected based on information in public databases such as in UCSC, Ensembl and NCBI genome browsers and others. Non-limiting examples of RNA transcripts for certain genes are listed in Table 1.

TABLE 1

Non-limiting examples of RNA transcripts for certain genes

GENE

SYMBOL
MRNA
SPECIES
GENE NAME

ABCA1
NM_013454

Mus

ATP-binding cassette, sub-family A (ABC1),

musculus

member 1

ABCA1
NM_005502

Homo

ATP-binding cassette, sub-family A (ABC1),

sapiens

member 1

ABCA4
NM_007378

Mus

ATP-binding cassette, sub-family A (ABC1),

musculus

member 4

ABCA4
NM_000350

Homo

ATP-binding cassette, sub-family A (ABC1),

sapiens

member 4

ABCB11
NM_003742

Homo

ATP-binding cassette, sub-family B

sapiens

(MDR/TAP), member 11

ABCB11
NM_021022

Mus

ATP-binding cassette, sub-family B

musculus

(MDR/TAP), member 11

ABCB4
NM_018850

Homo

ATP-binding cassette, sub-family B

sapiens

(MDR/TAP), member 4

ABCB4
NM_000443

Homo

ATP-binding cassette, sub-family B

sapiens

(MDR/TAP), member 4

ABCB4
NM_018849

Homo

ATP-binding cassette, sub-family B

sapiens

(MDR/TAP), member 4

ABCB4
NM_008830

Mus

ATP-binding cassette, sub-family B

musculus

(MDR/TAP), member 4

ABCG5
NM_022436

Homo

ATP-binding cassette, sub-family G (WHITE),

sapiens

member 5

ABCG5
NM_031884

Mus

ATP-binding cassette, sub-family G (WHITE),

musculus

member 5

ABCG8
NM_026180

Mus

ATP-binding cassette, sub-family G (WHITE),

musculus

member 8

ABCG8
NM_022437

Homo

ATP-binding cassette, sub-family G (WHITE),

sapiens

member 8

ADIPOQ
NM_009605

Mus

adiponectin, C1Q and collagen domain

musculus

containing

ADIPOQ
NM_004797

Homo

adiponectin, C1Q and collagen domain

sapiens

containing

ALB
NM_000477

Homo

albumin

sapiens

ALB
NM_009654

Mus

albumin

musculus

APOA1
NM_000039

Homo

apolipoprotein A-I

sapiens

APOA1
NM_009692

Mus

apolipoprotein A-I

musculus

APOE
NM_009696

Mus

apolipoprotein E

musculus

APOE
XM_001724655

Homo

hypothetical LOC100129500; apolipoprotein E

sapiens

APOE
XM_001722911

Homo

hypothetical LOC100129500; apolipoprotein E

sapiens

APOE
XM_001724653

Homo

hypothetical LOC100129500; apolipoprotein E

sapiens

APOE
NM_000041

Homo

hypothetical LOC100129500; apolipoprotein E

sapiens

APOE
XM_001722946

Homo

hypothetical LOC100129500; apolipoprotein E

sapiens

ATP2A2
NM_009722

Mus

ATPase, Ca++ transporting, cardiac muscle,

musculus

slow twitch 2

ATP2A2
NM_001110140

Mus

ATPase, Ca++ transporting, cardiac muscle,

musculus

slow twitch 2

ATP2A2
NM_001135765

Homo

ATPase, Ca++ transporting, cardiac muscle,

sapiens

slow twitch 2

ATP2A2
NM_170665

Homo

ATPase, Ca++ transporting, cardiac muscle,

sapiens

slow twitch 2

ATP2A2
NM_001681

Homo

ATPase, Ca++ transporting, cardiac muscle,

sapiens

slow twitch 2

BCL2L11
NM_006538

Homo

BCL2-like 11 (apoptosis facilitator)

sapiens

BCL2L11
NM_207002

Homo

BCL2-like 11 (apoptosis facilitator)

sapiens

BCL2L11
NM_138621

Homo

BCL2-like 11 (apoptosis facilitator)

sapiens

BCL2L11
NM_207680

Mus

BCL2-like 11 (apoptosis facilitator)

musculus

BCL2L11
NM_207681

Mus

BCL2-like 11 (apoptosis facilitator)

musculus

BCL2L11
NM_009754

Mus

BCL2-like 11 (apoptosis facilitator)

musculus

BDNF
NM_001143816

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001143815

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001143814

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001143813

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001143812

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001143806

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001143811

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001143805

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001143810

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001709

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_170735

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_170734

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_170733

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_170732

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_170731

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001143809

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001143807

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_001143808

Homo

brain-derived neurotrophic factor

sapiens

BDNF
NM_007540

Mus

brain derived neurotrophic factor

musculus

BDNF
NM_001048141

Mus

brain derived neurotrophic factor

musculus

BDNF
NM_001048142

Mus

brain derived neurotrophic factor

musculus

BDNF
NM_001048139

Mus

brain derived neurotrophic factor

musculus

BRCA1
NM_009764

Mus

breast cancer 1

musculus

BRCA1
NM_007296

Homo

breast cancer 1, early onset

sapiens

BRCA1
NM_007300

Homo

breast cancer 1, early onset

sapiens

BRCA1
NM_007297

Homo

breast cancer 1, early onset

sapiens

BRCA1
NM_007303

Homo

breast cancer 1, early onset

sapiens

BRCA1
NM_007298

Homo

breast cancer 1, early onset

sapiens

BRCA1
NM_007302

Homo

breast cancer 1, early onset

sapiens

BRCA1
NM_007299

Homo

breast cancer 1, early onset

sapiens

BRCA1
NM_007304

Homo

breast cancer 1, early onset

sapiens

BRCA1
NM_007294

Homo

breast cancer 1, early onset

sapiens

BRCA1
NM_007305

Homo

breast cancer 1, early onset

sapiens

BRCA1
NM_007295

Homo

breast cancer 1, early onset

sapiens

CD274
NM_014143

Homo

CD274 molecule

sapiens

CD274
NM_021893

Mus

CD274 antigen

musculus

CEP290
NM_025114

Homo

centrosomal protein 290 kDa

sapiens

CEP290
NM_146009

Mus

centrosomal protein 290

musculus

CFTR
NM_000492

Homo

cystic fibrosis transmembrane conductance

sapiens

regulator (ATP-binding cassette sub-family C,

member 7)

CFTR
NM_021050

Mus

cystic fibrosis transmembrane conductance

musculus

regulator homolog

EPO
NM_000799

Homo

erythropoietin

sapiens

EPO
NM_007942

Mus

erythropoietin

musculus

F7
NM_000131

Homo

coagulation factor VII (serum prothrombin

sapiens

conversion accelerator)

F7
NM_019616

Homo

coagulation factor VII (serum prothrombin

sapiens

conversion accelerator)

F7
NM_010172

Mus

coagulation factor VII

musculus

F8
NM_019863

Homo

coagulation factor VIII, procoagulant

sapiens

component

F8
NM_000132

Homo

coagulation factor VIII, procoagulant

sapiens

component

F8
NM_001161373

Mus

coagulation factor VIII

musculus

F8
NM_001161374

Mus

coagulation factor VIII

musculus

F8
NM_007977

Mus

coagulation factor VIII

musculus

FLI1
NM_002017

Homo

Friend leukemia virus integration 1

sapiens

FLI1
NM_001167681

Homo

Friend leukemia virus integration 1

sapiens

FLI1
NM_008026

Mus

Friend leukemia integration 1

musculus

FMR1
NM_008031

Mus

fragile X mental retardation syndrome 1

musculus

homolog

FMR1
NM_002024

Homo

fragile X mental retardation 1

sapiens

FNDC5
NM_001171941

Homo

fibronectin type III domain containing 5

sapiens

FNDC5
NM_153756

Homo

fibronectin type III domain containing 5

sapiens

FNDC5
NM_001171940

Homo

fibronectin type III domain containing 5

sapiens

FNDC5
NM_027402

Mus

fibronectin type III domain containing 5

musculus

FOXP3
NM_054039

Mus

forkhead box P3

musculus

FOXP3
NM_001114377

Homo

forkhead box P3

sapiens

FOXP3
NM_014009

Homo

forkhead box P3

sapiens

FXN
NM_001161706

Homo

frataxin

sapiens

FXN
NM_181425

Homo

frataxin

sapiens

FXN
NM_000144

Homo

frataxin

sapiens

FXN
NM_008044

Mus

frataxin

musculus

GCH1
NM_008102

Mus

GTP cyclohydrolase 1

musculus

GCH1
NM_000161

Homo

GTP cyclohydrolase 1

sapiens

GCH1
NM_001024070

Homo

GTP cyclohydrolase 1

sapiens

GCH1
NM_001024071

Homo

GTP cyclohydrolase 1

sapiens

GCH1
NM_001024024

Homo

GTP cyclohydrolase 1

sapiens

GCK
NM_010292

Mus

glucokinase

musculus

GCK
NM_000162

Homo

glucokinase (hexokinase 4)

sapiens

GCK
NM_033508

Homo

glucokinase (hexokinase 4)

sapiens

GCK
NM_033507

Homo

glucokinase (hexokinase 4)

sapiens

GLP1R
NM_021332

Mus

glucagon-like peptide 1 receptor; similar to

musculus

glucagon-like peptide-1 receptor

GLP1R
XM_001471951

Mus

glucagon-like peptide 1 receptor; similar to

musculus

glucagon-like peptide-1 receptor

GLP1R
NM_002062

Homo

glucagon-like peptide 1 receptor

sapiens

GRN
NM_002087

Homo

granulin

sapiens

GRN
NM_008175

Mus

granulin

musculus

HAMP
NM_021175

Homo

hepcidin antimicrobial peptide

sapiens

HAMP
NM_032541

Mus

hepcidin antimicrobial peptide

musculus

HBA2
NM_000517

Homo

hemoglobin, alpha 2; hemoglobin, alpha 1

sapiens

HBA2
NM_000558

Homo

hemoglobin, alpha 2; hemoglobin, alpha 1

sapiens

HBB
NM_000518

Homo

hemoglobin, beta

sapiens

HBB
XM_921413

Mus

hemoglobin beta chain complex

musculus

HBB
XM_903245

Mus

hemoglobin beta chain complex

musculus

HBB
XM_921395

Mus

hemoglobin beta chain complex

musculus

HBB
XM_903244

Mus

hemoglobin beta chain complex

musculus

HBB
XM_903246

Mus

hemoglobin beta chain complex

musculus

HBB
XM_909723

Mus

hemoglobin beta chain complex

musculus

HBB
XM_921422

Mus

hemoglobin beta chain complex

musculus

HBB
XM_489729

Mus

hemoglobin beta chain complex

musculus

HBB
XM_903242

Mus

hemoglobin beta chain complex

musculus

HBB
XM_903243

Mus

hemoglobin beta chain complex

musculus

HBB
XM_921400

Mus

hemoglobin beta chain complex

musculus

HBD
NM_000519

Homo

hemoglobin, delta

sapiens

HBE1
NM_005330

Homo

hemoglobin, epsilon 1

sapiens

HBG1
NM_000559

Homo

hemoglobin, gamma A

sapiens

HBG2
NM_000184

Homo

hemoglobin, gamma G

sapiens

HPRT1
NM_000194

Homo

hypoxanthine phosphoribosyltransferase 1

sapiens

IDO1
NM_008324

Mus

indoleamine 2,3-dioxygenase 1

musculus

IDO1
NM_002164

Homo

indoleamine 2,3-dioxygenase 1

sapiens

IGF1
NM_001111284

Homo

insulin-like growth factor 1 (somatomedin C)

sapiens

IGF1
NM_001111285

Homo

insulin-like growth factor 1 (somatomedin C)

sapiens

IGF1
NM_001111283

Homo

insulin-like growth factor 1 (somatomedin C)

sapiens

IGF1
NM_000618

Homo

insulin-like growth factor 1 (somatomedin C)

sapiens

IGF1
NM_001111274

Mus

insulin-like growth factor 1

musculus

IGF1
NM_010512

Mus

insulin-like growth factor 1

musculus

IGF1
NM_184052

Mus

insulin-like growth factor 1

musculus

IGF1
NM_001111276

Mus

insulin-like growth factor 1

musculus

IGF1
NM_001111275

Mus

insulin-like growth factor 1

musculus

IL10
NM_000572

Homo

interleukin 10

sapiens

IL10
NM_010548

Mus

interleukin 10

musculus

IL6
NM_031168

Mus

interleukin 6

musculus

IL6
NM_000600

Homo

interleukin 6 (interferon, beta 2)

sapiens

KCNMA1
NM_002247

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M, alpha

member 1

KCNMA1
NM_001161352

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M, alpha

member 1

KCNMA1
NM_001014797

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M, alpha

member 1

KCNMA1
NM_001161353

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M, alpha

member 1

KCNMA1
NM_010610

Mus

potassium large conductance calcium-

musculus

activated channel, subfamily M, alpha

member 1

KCNMB1
NM_031169

Mus

potassium large conductance calcium-

musculus

activated channel, subfamily M, beta

member 1

KCNMB1
NM_004137

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M, beta

member 1

KCNMB2
NM_028231

Mus

potassium large conductance calcium-

musculus

activated channel, subfamily M, beta

member 2

KCNMB2
NM_005832

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M, beta

member 2

KCNMB2
NM_181361

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M, beta

member 2

KCNMB3
NM_171829

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M beta member 3

KCNMB3
NM_171828

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M beta member 3

KCNMB3
NM_001163677

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M beta member 3

KCNMB3
NM_014407

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M beta member 3

KCNMB3
NM_171830

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M beta member 3

KCNMB3
XM_001475546

Mus

potassium large conductance calcium-

musculus

activated channel, subfamily M, beta

member 3

KCNMB3
XM_912348

Mus

potassium large conductance calcium-

musculus

activated channel, subfamily M, beta

member 3

KCNMB4
NM_021452

Mus

potassium large conductance calcium-

musculus

activated channel, subfamily M, beta

member 4

KCNMB4
NM_014505

Homo

potassium large conductance calcium-

sapiens

activated channel, subfamily M, beta

member 4

KLF1
NM_010635

Mus

Kruppel-like factor 1 (erythroid)

musculus

KLF1
NM_006563

Homo

Kruppel-like factor 1 (erythroid)

sapiens

KLF4
NM_010637

Mus

Kruppel-like factor 4 (gut)

musculus

KLF4
NM_004235

Homo

Kruppel-like factor 4 (gut)

sapiens

LAMA1
NM_005559.3

Homo

laminin, alpha 1

sapiens

LAMA1
NM_008480.2

Mus

laminin, alpha 1

musculus

LDLR
NM_000527

Homo

low density lipoprotein receptor

sapiens

LDLR
NM_010700

Mus

low density lipoprotein receptor

musculus

MBNL1
NM_021038.3,

Homo

muscleblind-like splicing regulator 1

NM_020007.3,

sapiens

NM_207293.1,

NM_207294.1,

NM_207295.1,

NM_207296.1,

NM_207297.1

MBNL1
NM_001253708.1,

Mus

muscleblind-like 1 (Drosophila)

NM_001253709.1,

musculus

NM_001253710.1,

NM_001253711.1,

NM_001253713.1,

NM_020007.3

MECP2
NM_010788

Mus

methyl CpG binding protein 2

musculus

MECP2
NM_001081979

Mus

methyl CpG binding protein 2

musculus

MECP2
NM_001110792

Homo

methyl CpG binding protein 2 (Rett

sapiens

syndrome)

MECP2
NM_004992

Homo

methyl CpG binding protein 2 (Rett

sapiens

syndrome)

MERTK
NM_006343.2

Homo

MER proto-oncogene, tyrosine kinase

sapiens

MERTK
NM_008587.1

Mus

c-mer proto-oncogene tyrosine kinase

musculus

MSX2
NM_013601

Mus

similar to homeobox protein; homeobox,

musculus

msh-like 2

MSX2
XM_001475886

Mus

similar to homeobox protein; homeobox,

musculus

msh-like 2

MSX2
NM_002449

Homo

msh homeobox 2

sapiens

MYBPC3
NM_008653

Mus

myosin binding protein C, cardiac

musculus

MYBPC3
NM_000256

Homo

myosin binding protein C, cardiac

sapiens

NANOG
NM_024865

Homo

Nanog homeobox pseudogene 8; Nanog

sapiens

homeobox

NANOG
XM_001471588

Mus

similar to Nanog homeobox; Nanog

musculus

homeobox

NANOG
NM_028016

Mus

similar to Nanog homeobox; Nanog

musculus

homeobox

NANOG
NM_001080945

Mus

similar to Nanog homeobox; Nanog

musculus

homeobox

NF1
NM_000267

Homo

neurofibromin 1

sapiens

NF1
NM_001042492

Homo

neurofibromin 1

sapiens

NF1
NM_001128147

Homo

neurofibromin 1

sapiens

NF1
NM_010897

Mus

neurofibromatosis 1

musculus

NKX2-1
NM_001079668

Homo

NK2 homeobox 1

sapiens

NKX2-1
NM_003317

Homo

NK2 homeobox 1

sapiens

NKX2-1
XM_002344771

Homo

NK2 homeobox 1

sapiens

NKX2-1
NM_009385

Mus

NK2 homeobox 1

musculus

NKX2-1
NM_001146198

Mus

NK2 homeobox 1

musculus

PAH
NM_008777

Mus

phenylalanine hydroxylase

musculus

PAH
NM_000277

Homo

phenylalanine hydroxylase

sapiens

PTEN
NM_000314

Homo

phosphatase and tensin homolog;

sapiens

phosphatase and tensin homolog

pseudogene 1

PTEN
NM_177096

Mus

phosphatase and tensin homolog

musculus

PTEN
NM_008960

Mus

phosphatase and tensin homolog

musculus

PTGS2
NM_011198

Mus

prostaglandin-endoperoxide synthase 2

musculus

PTGS2
NM_000963

Homo

prostaglandin-endoperoxide synthase 2

sapiens

(prostaglandin G/H synthase and

cyclooxygenase)

RB1
NM_009029

Mus

retinoblastoma 1

musculus

RB1
NM_000321

Homo

retinoblastoma 1

sapiens

RPS14
NM_020600

Mus

predicted gene 6204; ribosomal protein S14

musculus

RPS14
NM_001025071

Homo

ribosomal protein S14

sapiens

RPS14
NM_005617

Homo

ribosomal protein S14

sapiens

RPS14
NM_001025070

Homo

ribosomal protein S14

sapiens

RPS19
XM_204069

Mus

predicted gene 4327; predicted gene 8683;

musculus

similar to 40S ribosomal protein S19;

predicted gene 4510; predicted gene 13143;

predicted gene 9646; ribosomal protein S19;

predicted gene 9091; predicted gene 6636;

predicted gene 14072

RPS19
XM_991053

Mus

predicted gene 4327; predicted gene 8683;

musculus

similar to 40S ribosomal protein S19;

predicted gene 4510; predicted gene 13143;

predicted gene 9646; ribosomal protein S19;

predicted gene 9091; predicted gene 6636;

predicted gene 14072

RPS19
XM_905004

Mus

predicted gene 4327; predicted gene 8683;

musculus

similar to 40S ribosomal protein S19;

predicted gene 4510; predicted gene 13143;

predicted gene 9646; ribosomal protein S19;

predicted gene 9091; predicted gene 6636;

predicted gene 14072

RPS19
XM_001005575

Mus

predicted gene 4327; predicted gene 8683;

musculus

similar to 40S ribosomal protein S19;

predicted gene 4510; predicted gene 13143;

predicted gene 9646; ribosomal protein S19;

predicted gene 9091; predicted gene 6636;

predicted gene 14072

RPS19
NM_023133

Mus

predicted gene 4327; predicted gene 8683;

musculus

similar to 40S ribosomal protein S19;

predicted gene 4510; predicted gene 13143;

predicted gene 9646; ribosomal protein S19;

predicted gene 9091; predicted gene 6636;

predicted gene 14072

RPS19
XM_994263

Mus

predicted gene 4327; predicted gene 8683;

musculus

similar to 40S ribosomal protein S19;

predicted gene 4510; predicted gene 13143;

predicted gene 9646; ribosomal protein S19;

predicted gene 9091; predicted gene 6636;

predicted gene 14072

RPS19
XM_001481027

Mus

predicted gene 4327; predicted gene 8683;

musculus

similar to 40S ribosomal protein S19;

predicted gene 4510; predicted gene 13143;

predicted gene 9646; ribosomal protein S19;

predicted gene 9091; predicted gene 6636;

predicted gene 14072

RPS19
XM_913504

Mus

predicted gene 4327; predicted gene 8683;

musculus

similar to 40S ribosomal protein S19;

predicted gene 4510; predicted gene 13143;

predicted gene 9646; ribosomal protein S19;

predicted gene 9091; predicted gene 6636;

predicted gene 14072

RPS19
XM_001479631

Mus

predicted gene 4327; predicted gene 8683;

musculus

similar to 40S ribosomal protein S19;

predicted gene 4510; predicted gene 13143;

predicted gene 9646; ribosomal protein S19;

predicted gene 9091; predicted gene 6636;

predicted gene 14072

RPS19
XM_902221

Mus

predicted gene 4327; predicted gene 8683;

musculus

similar to 40S ribosomal protein S19;

predicted gene 4510; predicted gene 13143;

predicted gene 9646; ribosomal protein S19;

predicted gene 9091; predicted gene 6636;

predicted gene 14072

RPS19
XM_893968

Mus

predicted gene 4327; predicted gene 8683;

musculus

similar to 40S ribosomal protein S19;

predicted gene 4510; predicted gene 13143;

predicted gene 9646; ribosomal protein S19;

predicted gene 9091; predicted gene 6636;

predicted gene 14072

RPS19
NM_001022

Homo

ribosomal protein S19 pseudogene 3;

sapiens

ribosomal protein S19

SCARB1
NM_016741

Mus

scavenger receptor class B, member 1

musculus

SCARB1
NM_001082959

Homo

scavenger receptor class B, member 1

sapiens

SCARB1
NM_005505

Homo

scavenger receptor class B, member 1

sapiens

SERPINF1
NM_011340

Mus

serine (or cysteine) peptidase inhibitor, clade

musculus

F, member 1

SERPINF1
NM_002615

Homo

serpin peptidase inhibitor, clade F (alpha-2

sapiens

antiplasmin, pigment epithelium derived

factor), member 1

SIRT1
NM_001159590

Mus

sirtuin 1 (silent mating type information

musculus

regulation 2, homolog) 1 (S. cerevisiae)

SIRT1
NM_019812

Mus

sirtuin 1 (silent mating type information

musculus

regulation 2, homolog) 1 (S. cerevisiae)

SIRT1
NM_001159589

Mus

sirtuin 1 (silent mating type information

musculus

regulation 2, homolog) 1 (S. cerevisiae)

SIRT1
NM_012238

Homo

sirtuin (silent mating type information

sapiens

regulation 2 homolog) 1 (S. cerevisiae)

SIRT1
NM_001142498

Homo

sirtuin (silent mating type information

sapiens

regulation 2 homolog) 1 (S. cerevisiae)

SIRT6
NM_016539

Homo

sirtuin (silent mating type information

sapiens

regulation 2 homolog) 6 (S. cerevisiae)

SIRT6
NM_001163430

Mus

sirtuin 6 (silent mating type information

musculus

regulation 2, homolog) 6 (S. cerevisiae)

SIRT6
NM_181586

Mus

sirtuin 6 (silent mating type information

musculus

regulation 2, homolog) 6 (S. cerevisiae)

SMAD7
NM_005904

Homo

SMAD family member 7

sapiens

SMAD7
NM_001042660

Mus

MAD homolog 7 (Drosophila)

musculus

SMN1
NM_000344.3

Homo

Survival Motor Neuron 1

sapiens

SMN1
NM_022874.2

Homo

Survival Motor Neuron 1

sapiens

SMN2
NM_017411.3

Homo

Survival Motor Neuron 2

NM_022875.2

sapiens

NM_022876.2

NM_022877.2

SSPN
NM_001135823.1,

Homo

sarcospan

NM_005086.4

sapiens

SSPN
NM_010656.2

Homo

sarcospan

sapiens

ST7
NM_021908

Homo

suppression of tumorigenicity 7

sapiens

ST7
NM_018412

Homo

suppression of tumorigenicity 7

sapiens

STAT3
NM_213660

Mus

similar to Stat3B; signal transducer and

musculus

activator of transcription 3

STAT3
XM_001474017

Mus

similar to Stat3B; signal transducer and

musculus

activator of transcription 3

STAT3
NM_213659

Mus

similar to Stat3B; signal transducer and

musculus

activator of transcription 3

STAT3
NM_011486

Mus

similar to Stat3B; signal transducer and

musculus

activator of transcription 3

STAT3
NM_213662

Homo

signal transducer and activator of

sapiens

transcription 3 (acute-phase response factor)

STAT3
NM_003150

Homo

signal transducer and activator of

sapiens

transcription 3 (acute-phase response factor)

STAT3
NM_139276

Homo

signal transducer and activator of

sapiens

transcription 3 (acute-phase response factor)

UTRN
NM_007124

Homo

utrophin

sapiens

UTRN
NM_011682

Mus

utrophin

musculus

NFE2L2
NM_001145412.2,

Homo

nuclear factor, erythroid 2-like 2

NM_001145413.2,

sapiens

NM_006164.4

NFE2L2
NM_010902.3

Mus

nuclear factor, erythroid 2-like 2

musculus

ACTB
NM_001101.3

Homo

actin, beta

sapiens

ACTB
NM_007393.3

Mus

actin, beta

musculus

ANRIL
NR_003529.3,

Homo

CDKN2B antisense RNA 1 (also called

NR_047532.1,

sapiens

CDKN2B)

NR_047533.1,

NR_047534.1,

NR_047535.1,

NR_047536.1,

NR_047538.1,

NR_047539.1,

NR_047540.1,

NR_047541.1,

NR_047542.1,

NR_047543.1

HOTAIR
NR_003716.3,

Homo

HOX transcript antisense RNA

NR_047517.1,

sapiens

NR_047518.1

HOTAIR
NR_047528.1

Mus

HOX transcript antisense RNA

musculus

DINO
JX993265

Homo

Damage Induced NOncoding

sapiens

DINO
JX993266

Mus

Damage Induced NOncoding

musculus

HOTTIP
NR_037843.3

Homo

HOXA distal transcript antisense RNA

sapiens

HOTTIP
NR_110441.1,

Mus

Hoxa distal transcript antisense RNA

NR_110442.1

musculus

NEST
NR_104124.1

Homo

Homo sapiens IFNG antisense RNA 1 (IFNG-

sapiens

AS1), transcript variant 1, long non-coding

RNA.

NEST
NR_104123.1

Mus

Theiler's murine encephalomyelitis virus

musculus

persistence candidate gene 1

Synthetic RNAs

In some aspects of the disclosure, formulations and methods are provided for stabilizing a synthetic RNA (e.g., a synthetic RNA that is to be delivered to a cell). In some embodiments, the formulations and methods involve contacting a synthetic RNA with one or more stabilizing oligonucleotides that bind to a 5′ region of the synthetic RNA and a 3′ region of the synthetic RNA and that when bound to the synthetic RNA form a circularized product with the synthetic RNA. In some embodiments, the synthetic RNA is contacted with the one or more stabilizing oligonucleotides outside of a cell.

In some embodiments, synthetic RNAs are provided with chemistries suitable for delivery, hybridization and stability within cells which are targeted and stabilized by a stabilizing oligonucleotide, e.g., as described herein. Furthermore, in some embodiments, synthetic RNA chemistries are provided that are useful for controlling the pharmacokinetics, biodistribution, bioavailability and/or efficacy of the synthetic RNAs. Accordingly, synthetic RNAs described herein may be modified, e.g., comprise a modified sugar moiety, a modified internucleoside linkage, a modified nucleotide and/or combinations thereof. In addition, the synthetic RNAs may exhibit one or more of the following properties: do not induce substantial cleavage or degradation of the target RNA and/or stabilizing oligonucleotide; do not cause substantially complete cleavage or degradation of the target RNA and/or stabilizing oligonucleotide; do not activate the RNAse H pathway; do not activate RISC; do not recruit any Argonaute family protein; are not cleaved by Dicer; do not mediate alternative splicing; are not immune stimulatory; are nuclease resistant; have improved cell uptake compared to unmodified synthetic RNAs; are not toxic to cells or mammals; and may have improved endosomal exit.

Methods for Modulating Gene Expression

In one aspect, the disclosure relates to methods for modulating (e.g., increasing) stability of RNA transcripts in cells. The cells can be in vitro, ex vivo, or in vivo. The cells can be in a subject who has a disease resulting from reduced expression or activity of the RNA transcript or its corresponding protein product in the case of mRNAs. In some embodiments, methods for modulating stability of RNA transcripts in cells comprise delivering to the cell a formulation comprising a stabilizing oligonucleotide and a particle, e.g., nanoparticle. In some embodiments, delivery of a formulation comprising a stabilizing oligonucleotide and a particle as described herein to the cell results in an increase in stability of a target RNA that is at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200% or more greater than a level of stability of the target RNA in a control cell. An appropriate control cell may be a cell to which a stabilizing oligonucleotide has not been delivered or to which a negative control has been delivered (e.g., a scrambled oligo, a carrier, etc.).

Another aspect of the disclosure provides methods of treating a disease or condition associated with low levels of a particular RNA in a subject. Accordingly, in some embodiments, methods are provided that comprise administering to a subject (e.g. a human) a formulation comprising a stabilizing oligonucleotide and a particle as described herein to increase mRNA stability in cells of the subject for purposes of increasing protein levels. In some embodiments, the increase in protein levels is at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, or more, higher than the amount of a protein in the subject (e.g., in a cell or tissue of the subject) before administering or in a control subject which has not been administered the stabilizing oligonucleotide or that has been administered a negative control (e.g., a scrambled oligo, a carrier, etc.). In some embodiments, methods are provided that comprise administering to a subject (e.g. a human) a formulation comprising a stabilizing oligonucleotide and a particle as described herein to increase stability of non-coding RNAs in cells of the subject for purposes of increasing activity of those non-coding RNAs.

A subject can include a non-human mammal, e.g. mouse, rat, guinea pig, rabbit, cat, dog, goat, cow, or horse. In preferred embodiments, a subject is a human. Stabilizing oligonucleotides may be employed as therapeutic moieties in the treatment of disease states in animals, including humans. Stabilizing oligonucleotides can be useful therapeutic modalities that can be configured to be useful in treatment regimes for the treatment of cells, tissues and animals, especially humans.

For therapeutics, an animal, preferably a human, suspected of having a disease associated with low levels of an RNA or protein is treated by administering a formulation comprising a stabilizing oligonucleotide and a particle in accordance with this disclosure. For example, in one non-limiting embodiment, the methods comprise the step of administering to the animal in need of treatment, a therapeutically effective amount of an formulation as described herein. Table 2 listed examples of diseases or conditions that may be treated by targeting mRNA transcripts with stabilizing oligonucleotides. In some embodiments, cells used in the methods disclosed herein may, for example, be cells obtained from a subject having one or more of the conditions listed in Table 2, or from a subject that is a disease model of one or more of the conditions listed in Table 2.

TABLE 2

Examples of diseases or conditions treatable with oligonucleotides targeting

associated mRNA.

Gene
Disease or conditions

FXN
Friedreich's Ataxia

SMN
Spinal muscular atrophy (SMA) types I-IV, Amyotrophic Lateral

Sclerosis (ALS)

UTRN
Muscular dystrophy (MD) (e.g., Duchenne's muscular dystrophy,

Becker's muscular dystrophy, myotonic dystrophy)

HEMOGLOBIN
Anemia, microcytic anemia, sickle cell anemia and/or thalassemia (e.g.,

alpha-thalassemia, beta-thalaseemia, delta-thalessemia), beta-thalaseemia

(e.g., thalassemia minor/intermedia/major)

ATP2A2
Cardiac conditions (e.g., congenital heart disease, aortic aneurysms,

aortic dissections, arrhythmia, cardiomyopathy, and congestive heart

failure), Darier-White disease and Acrokeratosis verruciformi

APOA1/
Dyslipidemia (e.g. Hyperlipidemia) and atherosclerosis (e.g. coronary

ABCA1
artery disease (CAD) and myocardial infarction (MI))

PTEN
Cancer, such as, leukemias, lymphomas, myelomas, carcinomas,

metastatic carcinomas, sarcomas, adenomas, nervous system cancers and

genito-urinary cancers. In some embodiments, the cancer is adult and

pediatric acute lymphoblastic leukemia, acute myeloid leukemia,

adrenocortical carcinoma, AIDS-related cancers, anal cancer, cancer of

the appendix, astrocytoma, basal cell carcinoma, bile duct cancer,

bladder cancer, bone cancer, osteosarcoma, fibrous histiocytoma, brain

cancer, brain stem glioma, cerebellar astrocytoma, malignant glioma,

ependymoma, medulloblastoma, supratentorial primitive

neuroectodermal tumors, hypothalamic glioma, breast cancer, male

breast cancer, bronchial adenomas, Burkitt lymphoma, carcinoid tumor,

carcinoma of unknown origin, central nervous system lymphoma,

cerebellar astrocytoma, malignant glioma, cervical cancer, childhood

cancers, chronic lymphocytic leukemia, chronic myelogenous leukemia,

chronic myeloproliferative disorders, colorectal cancer, cutaneous T-cell

lymphoma, endometrial cancer, ependymoma, esophageal cancer, Ewing

family tumors, extracranial germ cell tumor, extragonadal germ cell

tumor, extrahepatic bile duct cancer, intraocular melanoma,

retinoblastoma, gallbladder cancer, gastric cancer, gastrointestinal

stromal tumor, extracranial germ cell tumor, extragonadal germ cell

tumor, ovarian germ cell tumor, gestational trophoblastic tumor, glioma,

hairy cell leukemia, head and neck cancer, hepatocellular cancer,

Hodgkin lymphoma, non-Hodgkin lymphoma, hypopharyngeal cancer,

hypothalamic and visual pathway glioma, intraocular melanoma, islet

cell tumors, Kaposi sarcoma, kidney cancer, renal cell cancer, laryngeal

cancer, lip and oral cavity cancer, small cell lung cancer, non-small cell

lung cancer, primary central nervous system lymphoma, Waldenstrom

macroglobulinema, malignant fibrous histiocytoma, medulloblastoma,

melanoma, Merkel cell carcinoma, malignant mesothelioma, squamous

neck cancer, multiple endocrine neoplasia syndrome, multiple myeloma,

mycosis fungoides, myelodysplastic syndromes, myeloproliferative

disorders, chronic myeloproliferative disorders, nasal cavity and

paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma,

oropharyngeal cancer, ovarian cancer, pancreatic cancer, parathyroid

cancer, penile cancer, pharyngeal cancer, pheochromocytoma,

pineoblastoma and supratentorial primitive neuroectodermal tumors,

pituitary cancer, plasma cell neoplasms, pleuropulmonary blastoma,

prostate cancer, rectal cancer, rhabdomyosarcoma, salivary gland cancer,

soft tissue sarcoma, uterine sarcoma, Sezary syndrome, non-melanoma

skin cancer, small intestine cancer, squamous cell carcinoma, squamous

neck cancer, supratentorial primitive neuroectodermal tumors, testicular

cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer,

transitional cell cancer, trophoblastic tumors, urethral cancer, uterine

cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or Wilms tumor

BDNF
Amyotrophic lateral sclerosis (ALS, also known as Lou Gehrig's

disease), Alzheimer's Disease (AD), and Parkinson's Disease (PD),

Neurodegeneration

MECP2
Rett Syndrome, MECP2-related severe neonatal encephalopathy,

Angelman syndrome, or PPM-X syndrome

FOXP3
Diseases or disorders associated with aberrant immune cell (e.g., T cell)

activation, e.g., autoimmune or inflammatory diseases or disorders.

Examples of autoimmune diseases and disorders that may be treated

according to the methods disclosed herein include, but are not limited to,

Acute Disseminated Encephalomyelitis (ADEM), Acute necrotizing

hemorrhagic leukoencephalitis, Addison's disease,

Agammaglobulinemia, Alopecia areata, Amyloidosis, Ankylosing

spondylitis, Anti-GBM/Anti-TBM nephritis, Antiphospholipid syndrome

(APS), Autoimmune angioedema, Autoimmune aplastic anemia,

Autoimmune dysautonomia, Autoimmune hepatitis, Autoimmune

hyperlipidemia, Autoimmune immunodeficiency, Autoimmune inner ear

disease (AIED), Autoimmune myocarditis, Autoimmune oophoritis,

Autoimmune pancreatitis, Autoimmune retinopathy, Autoimmune

thrombocytopenic purpura (ATP), Autoimmune thyroid disease,

Autoimmune urticaria, Axonal & neuronal neuropathies, Balo disease,

Behcet's disease, Bullous pemphigoid, Cardiomyopathy, Castleman

disease, Celiac disease, Chagas disease, Chronic inflammatory

demyelinating polyneuropathy (CIDP), Chronic recurrent multifocal

ostomyelitis (CRMO), Churg-Strauss syndrome, Cicatricial

pemphigoid/benign mucosal pemphigoid, inflammatory bowel disease

(e.g., Crohn's disease or Ulcerative colitis), Cogans syndrome, Cold

agglutinin disease, Congenital heart block, Coxsackie myocarditis,

CREST disease, Essential mixed cryoglobulinemia, Demyelinating

neuropathies, Dermatitis herpetiformis, Dermatomyositis, Devic's

disease (neuromyelitis optica), Discoid lupus, Dressler's syndrome,

Endometriosis, Eosinophilic esophagitis, Eosinophilic fasciitis, Erythema

nodosum, Experimental allergic encephalomyelitis, Evans syndrome,

Fibrosing alveolitis, Giant cell arteritis (temporal arteritis), Giant cell

myocarditis, Glomerulonephritis, Goodpasture's syndrome,

Granulomatosis with Polyangiitis (GPA) (formerly called Wegener's

Granulomatosis), Graves' disease, Guillain-Barre syndrome, Hashimoto's

encephalitis, Hashimoto's thyroiditis, Hemolytic anemia, Henoch-

Schonlein purpura, Herpes gestationis, Hypogammaglobulinemia,

Idiopathic thrombocytopenic purpura (ITP), IgA nephropathy, IgG4-

related sclerosing disease, Immunoregulatory lipoproteins, Inclusion

body myositis, Interstitial cystitis, IPEX (Immunodysregulation,

Polyendocrinopathy, and Enteropathy, X-linked) syndrome, Juvenile

arthritis, Juvenile diabetes (Type 1 diabetes), Juvenile myositis,

Kawasaki syndrome, Lambert-Eaton syndrome, Leukocytoclastic

vasculitis, Lichen planus, Lichen sclerosus, Ligneous conjunctivitis,

Linear IgA disease (LAD), systemic lupus erythematosus (SLE), chronic

Lyme disease, Meniere's disease, Microscopic polyangiitis, Mixed

connective tissue disease (MCTD), Mooren's ulcer, Mucha-Habermann

disease, Multiple sclerosis, Myasthenia gravis, Myositis, Narcolepsy,

Neuromyelitis optica (Devic's), Neutropenia, Ocular cicatricial

pemphigoid, Optic neuritis, Palindromic rheumatism, PANDAS

(Pediatric Autoimmune Neuropsychiatric Disorders Associated with

Streptococcus), Paraneoplastic cerebellar degeneration, Paroxysmal

nocturnal hemoglobinuria (PNH), Parry Romberg syndrome,

Parsonnage-Turner syndrome, Pars planitis (peripheral uveitis),

Pemphigus, Peripheral neuropathy, Perivenous encephalomyelitis,

Pernicious anemia, POEMS syndrome, Polyarteritis nodosa, Type I, II,

& III autoimmune polyglandular syndromes, Polymyalgia rheumatica,

Polymyositis, Postmyocardial infarction syndrome, Postpericardiotomy

syndrome, Progesterone dermatitis, Primary biliary cirrhosis, Primary

sclerosing cholangitis, Psoriasis, Psoriatic arthritis, Idiopathic pulmonary

fibrosis, Pyoderma gangrenosum, Pure red cell aplasia, Raynauds

phenomenon, Reactive Arthritis, Reflex sympathetic dystrophy, Reiter's

syndrome, Relapsing polychondritis, Restless legs syndrome,

Retroperitoneal fibrosis, Rheumatic fever, Rheumatoid arthritis,

Sarcoidosis, Schmidt syndrome, Scleritis, Scleroderma, Sjogren's

syndrome, Sperm & testicular autoimmunity, Stiff person syndrome,

Subacute bacterial endocarditis (SBE), Susac's syndrome, Sympathetic

ophthalmia, Takayasu's arteritis, Temporal arteritis/Giant cell arteritis,

Thrombocytopenic purpura (TTP), Tolosa-Hunt syndrome, Transverse

myelitis, Type 1 diabetes, Undifferentiated connective tissue disease

(UCTD), Uveitis, Vasculitis, Vesiculobullous dermatosis, Vitiligo, and

Wegener's granulomatosis (also called Granulomatosis with Polyangiitis

(GPA)). Further examples of autoimmune disease or disorder include

inflammatory bowel disease (e.g., Crohn's disease or Ulcerative colitis),

IPEX syndrome, Multiple sclerosis, Psoriasis, Rheumatoid arthritis, SLE

or Type 1 diabetes. Examples of inflammatory diseases or disorders that

may be treated according to the methods disclosed herein include, but are

not limited to, Acne Vulgaris, Appendicitis, Arthritis, Asthma,

Atherosclerosis, Allergies (Type 1 Hypersensitivity), Bursitis, Colitis,

Chronic Prostatitis, Cystitis, Dermatitis, Glomerulonephritis,

Inflammatory Bowel Disease, Inflammatory Myopathy (e.g.,

Polymyositis, Dermatomyositis, or Inclusion-body Myositis),

Inflammatory Lung Disease, Interstitial Cystitis, Meningitis, Pelvic

Inflammatory Disease, Phlebitis, Psoriasis, Reperfusion Injury,

Rheumatoid Arthritis, Sarcoidosis, Tendonitis, Tonsilitis, Transplant

Rejection, and Vasculitis. In some embodiments, the inflammatory

disease or disorder is asthma.

Route of Delivery

A formulation comprising a stabilizing oligonucleotide and a particle may be delivered to a subject by a variety of routes. Exemplary routes include: intravenous, intradermal, topical, rectal, parenteral, anal, intravaginal, intranasal, pulmonary, ocular. The term “therapeutically effective amount” is the amount of stabilizing oligonucleotide present in the composition that is needed to provide the desired level of gene expression (e.g., by stabilizing RNA transcripts) in the subject to be treated to give the anticipated physiological response. The term “physiologically effective amount” is that amount delivered to a subject to give the desired palliative or curative effect. The term “pharmaceutically acceptable carrier” means that the carrier can be administered to a subject with no significant adverse toxicological effects to the subject.

In some embodiments, formulations comprising stabilizing oligonucleotide molecules and particles of the disclosure can be incorporated into pharmaceutical compositions suitable for administration. In some embodiments, such formulations may include one or more species of stabilizing oligonucleotide and a particle, and a pharmaceutically acceptable carrier, e.g., to the extent such a carrier is compatible with desired oligonucleotide molecules and particles interactions. As used herein the language “pharmaceutically acceptable carrier” is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.

The formulations of the present disclosure may be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be topical (including ophthalmic, vaginal, rectal, intranasal, transdermal), oral or parenteral. Parenteral administration includes intravenous drip, subcutaneous, intraperitoneal or intramuscular injection, or intrathecal or intraventricular administration.

The route and site of administration may be chosen to enhance targeting. For example, to target muscle cells, intramuscular injection into the muscles of interest would be a logical choice. Lung cells might be targeted by administering a formulation in aerosol form. The vascular endothelial cells could be targeted by coating a balloon catheter with a formulation of the present disclosure and mechanically introducing the oligonucleotide.

Topical administration refers to the delivery to a subject by contacting the formulation directly to a surface of the subject. The most common form of topical delivery is to the skin, but a formulation disclosed herein can also be directly applied to other surfaces of the body, e.g., to the eye, a mucous membrane, to surfaces of a body cavity or to an internal surface. As mentioned above, the most common topical delivery is to the skin. The term encompasses several routes of administration including, but not limited to, topical and transdermal. These modes of administration typically include penetration of the skin's permeability barrier and efficient delivery to the target tissue or stratum. Topical administration can be used as a means to penetrate the epidermis and dermis and ultimately achieve systemic delivery of the composition. Topical administration can also be used as a means to selectively deliver oligonucleotides to the epidermis or dermis of a subject, or to specific strata thereof, or to an underlying tissue.

Formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable. Coated condoms, gloves and the like may also be useful.

Transdermal delivery is a valuable route for the administration of lipid soluble therapeutics. The dermis is more permeable than the epidermis and therefore absorption is much more rapid through abraded, burned or denuded skin. Inflammation and other physiologic conditions that increase blood flow to the skin also enhance transdermal adsorption. Absorption via this route may be enhanced by the use of an oily vehicle (inunction) or through the use of one or more penetration enhancers. Other effective ways to deliver a formulation disclosed herein via the transdermal route include hydration of the skin and the use of controlled release topical patches. The transdermal route provides a potentially effective means to deliver a formulation disclosed herein for systemic and/or local therapy. In addition, iontophoresis (transfer of ionic solutes through biological membranes under the influence of an electric field), phonophoresis or sonophoresis (use of ultrasound to enhance the absorption of various therapeutic agents across biological membranes, notably the skin and the cornea), and optimization of vehicle characteristics relative to dose position and retention at the site of administration may be useful methods for enhancing the transport of topically applied compositions across skin and mucosal sites.

Both the oral and nasal membranes offer advantages over other routes of administration. For example, formulations administered through these membranes may have a rapid onset of action, provide therapeutic plasma levels, avoid first pass effect of hepatic metabolism, and avoid exposure of the oligonucleotides to the hostile gastrointestinal (GI) environment. Additional advantages include easy access to the membrane sites so that the formulation can be applied, localized and removed easily.

In oral delivery, formulations can be targeted to a surface of the oral cavity, e.g., to sublingual mucosa which includes the membrane of ventral surface of the tongue and the floor of the mouth or the buccal mucosa which constitutes the lining of the cheek. The sublingual mucosa is relatively permeable thus giving rapid absorption and acceptable bioavailability of many agents. Further, the sublingual mucosa is convenient, acceptable and easily accessible.

A formulation may also be administered to the buccal cavity of a human being by spraying into the cavity, without inhalation, from a metered dose spray dispenser, a mixed micellar pharmaceutical formulation as described above and a propellant. In one embodiment, the dispenser is first shaken prior to spraying the pharmaceutical formulation and propellant into the buccal cavity.

Formulations for oral administration include powders or granules, suspensions or solutions in water, syrups, slurries, emulsions, elixirs or non-aqueous media, tablets, capsules, lozenges, or troches. In the case of tablets, carriers that can be used include lactose, sodium citrate and salts of phosphoric acid. Various disintegrants such as starch, and lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc, are commonly used in tablets. For oral administration in capsule form, useful diluents are lactose and high molecular weight polyethylene glycols. When aqueous suspensions are required for oral use, the nucleic acid compositions can be combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring agents can be added.

Parenteral administration includes intravenous drip, subcutaneous, intraperitoneal or intramuscular injection, intrathecal or intraventricular administration. In some embodiments, parental administration involves administration directly to the site of disease (e.g. injection into a tumor).

Formulations for parenteral administration may include sterile aqueous solutions which may also contain buffers, diluents and other suitable additives. Intraventricular injection may be facilitated by an intraventricular catheter, for example, attached to a reservoir. For intravenous use, the total concentration of solutes should be controlled to render the preparation isotonic.

Any of the formulations described herein can be administered to ocular tissue. For example, the formulations can be applied to the surface of the eye or nearby tissue, e.g., the inside of the eyelid. For ocular administration, ointments or droppable liquids may be delivered by ocular delivery systems known to the art such as applicators or eye droppers. Such formulations can include mucomimetics such as hyaluronic acid, chondroitin sulfate, hydroxypropyl methylcellulose or poly(vinyl alcohol), preservatives such as sorbic acid, EDTA or benzylchronium chloride, and the usual quantities of diluents and/or carriers. A formulation can also be administered to the interior of the eye, and can be introduced by a needle or other delivery device which can introduce it to a selected area or structure.

Pulmonary delivery formulations can be delivered by inhalation by the patient of a dispersion so that the composition, preferably oligonucleotides, within the dispersion can reach the lung where it can be readily absorbed through the alveolar region directly into blood circulation. Pulmonary delivery can be effective both for systemic delivery and for localized delivery to treat diseases of the lungs.

Pulmonary delivery can be achieved by different approaches, including the use of nebulized, aerosolized, micellular and dry powder-based formulations. Delivery can be achieved with liquid nebulizers, aerosol-based inhalers, and dry powder dispersion devices. Metered-dose devices are preferred. One of the benefits of using an atomizer or inhaler is that the potential for contamination is minimized because the devices are self-contained. Dry powder dispersion devices, for example, deliver agents that may be readily formulated as dry powders. A formulation may be stably stored as lyophilized or spray-dried powders by itself or in combination with suitable powder carriers. The delivery of a composition for inhalation can be mediated by a dosing timing element which can include a timer, a dose counter, time measuring device, or a time indicator which when incorporated into the device enables dose tracking, compliance monitoring, and/or dose triggering to a patient during administration of the aerosol medicament.

The term “powder” means a composition that consists of finely dispersed solid particles that are free flowing and capable of being readily dispersed in an inhalation device and subsequently inhaled by a subject so that the particles reach the lungs to permit penetration into the alveoli. Thus, the powder is said to be “respirable.” Preferably the average particle size is less than about 10 am in diameter preferably with a relatively uniform spheroidal shape distribution. More preferably the diameter is less than about 7.5 am and most preferably less than about 5.0 μm. Usually the particle size distribution is between about 0.1 μm and about 5 μm in diameter, particularly about 0.3 μm to about 5 μm.

The term “dry” means that the formulation has a moisture content below about 10% by weight (% w) water, usually below about 5% w and preferably less it than about 3% w. A dry composition can be such that the particles are readily dispersible in an inhalation device to form an aerosol.

The types of pharmaceutical excipients that are useful as carrier include stabilizers such as human serum albumin (HSA), bulking agents such as carbohydrates, amino acids and polypeptides; pH adjusters or buffers; salts such as sodium chloride; and the like. These carriers may be in a crystalline or amorphous form or may be a mixture of the two.

Suitable pH adjusters or buffers include organic salts prepared from organic acids and bases, such as sodium citrate, sodium ascorbate, and the like; sodium citrate is preferred. Pulmonary administration of a micellar oligonucleotide formulation may be achieved through metered dose spray devices with propellants such as tetrafluoroethane, heptafluoroethane, dimethylfluoropropane, tetrafluoropropane, butane, isobutane, dimethyl ether and other non-CFC and CFC propellants.

Exemplary devices include devices which are introduced into the vasculature, e.g., devices inserted into the lumen of a vascular tissue, or which devices themselves form a part of the vasculature, including stents, catheters, heart valves, and other vascular devices. These devices, e.g., catheters or stents, can be placed in the vasculature of the lung, heart, or leg.

Other devices include non-vascular devices, e.g., devices implanted in the peritoneum, or in organ or glandular tissue, e.g., artificial organs. The device can release a therapeutic substance in addition to an oligonucleotide formulation, e.g., a device can release insulin.

In one embodiment, unit doses or measured doses of a formulation are dispensed by an implanted device. The device can include a sensor that monitors a parameter within a subject. For example, the device can include pump, e.g., and, optionally, associated electronics.

Tissue, e.g., cells or organs can be treated with a formulation, ex vivo and then administered or implanted in a subject. The tissue can be autologous, allogeneic, or xenogeneic tissue. E.g., tissue can be treated to reduce graft v. host disease. In other embodiments, the tissue is allogeneic and the tissue is treated to treat a disorder characterized by unwanted gene expression in that tissue. E.g., tissue, e.g., hematopoietic cells, e.g., bone marrow hematopoietic cells, can be treated to inhibit unwanted cell proliferation. Introduction of treated tissue, whether autologous or transplant, can be combined with other therapies. In some implementations, oligonucleotide treated cells are insulated from other cells, e.g., by a semi-permeable porous barrier that prevents the cells from leaving the implant, but enables molecules from the body to reach the cells and molecules produced by the cells to enter the body. In one embodiment, the porous barrier is formed from alginate.

In one embodiment, a contraceptive device is coated with or contains a formulation comprising a stabilizing oligonucleotide and a particle. Exemplary devices include condoms, diaphragms, IUD (implantable uterine devices, sponges, vaginal sheaths, and birth control devices.

Dosage

In one aspect, the disclosure features a method of administering a formulation comprising a stabilizing oligonucleotide and a particle to a subject (e.g., a human subject). In one embodiment, the unit dose is between about 10 mg and 25 mg per kg of bodyweight. In one embodiment, the unit dose is between about 1 mg and 100 mg per kg of bodyweight. In one embodiment, the unit dose is between about 0.1 mg and 500 mg per kg of bodyweight. In some embodiments, the unit dose is more than 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 2, 5, 10, 25, 50 or 100 mg per kg of bodyweight.

The defined amount can be an amount effective to treat or prevent a disease or disorder, e.g., a disease or disorder associated with low levels of an RNA or protein. The unit dose, for example, can be administered by injection (e.g., intravenous or intramuscular), an inhaled dose, or a topical application.

In some embodiments, the unit dose is administered daily. In some embodiments, less frequently than once a day, e.g., less than every 2, 4, 8 or 30 days. In another embodiment, the unit dose is not administered with a frequency (e.g., not a regular frequency). For example, the unit dose may be administered a single time. In some embodiments, the unit dose is administered more than once a day, e.g., once an hour, two hours, four hours, eight hours, twelve hours, etc.

In one embodiment, a subject is administered an initial dose and one or more maintenance doses of a formulation comprising a stabilizing oligonucleotide and a particle. The maintenance dose or doses are generally lower than the initial dose, e.g., one-half less of the initial dose. A maintenance regimen can include treating the subject with a dose or doses ranging from 0.0001 to 100 mg/kg of body weight per day, e.g., 100, 10, 1, 0.1, 0.01, 0.001, or 0.0001 mg per kg of bodyweight per day. The maintenance doses may be administered no more than once every 1, 5, 10, or 30 days. Further, the treatment regimen may last for a period of time which will vary depending upon the nature of the particular disease, its severity and the overall condition of the patient. In some embodiments the dosage may be delivered no more than once per day, e.g., no more than once per 24, 36, 48, or more hours, e.g., no more than once for every 5 or 8 days. Following treatment, the patient can be monitored for changes in his condition and for alleviation of the symptoms of the disease state. The dosage of the formulation may either be increased in the event the patient does not respond significantly to current dosage levels, or the dose may be decreased if an alleviation of the symptoms of the disease state is observed, if the disease state has been ablated, or if undesired side-effects are observed.

The effective dose can be administered in a single dose or in two or more doses, as desired or considered appropriate under the specific circumstances. If desired to facilitate repeated or frequent infusions, implantation of a delivery device, e.g., a pump, semi-permanent stent (e.g., intravenous, intraperitoneal, intracisternal or intracapsular), or reservoir may be advisable.

In some cases, a patient is treated with a formulation comprising a stabilizing oligonucleotide and a particle in conjunction with other therapeutic modalities.

Following successful treatment, it may be desirable to have the patient undergo maintenance therapy to prevent the recurrence of the disease state, wherein the formulation of the disclosure is administered in maintenance doses, ranging from 0.0001 mg to 100 mg per kg of body weight.

The concentration of an oligonucleotide formulation is an amount sufficient to be effective in treating or preventing a disorder or to regulate a physiological condition in humans. The concentration or amount of formulation administered will depend on the parameters determined for the agent and the method of administration, e.g. nasal, buccal, pulmonary. For example, nasal formulations may tend to require much lower concentrations of some ingredients in order to avoid irritation or burning of the nasal passages. It is sometimes desirable to dilute an oral formulation up to 10-100 times in order to provide a suitable nasal formulation.

Certain factors may influence the dosage required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of an oligonucleotide in a formulation can include a single treatment or, preferably, can include a series of treatments. It will also be appreciated that the effective dosage of an oligonucleotide in a formulation used for treatment may increase or decrease over the course of a particular treatment. For example, the subject can be monitored after administering a formulation comprising a stabilizing oligonucleotide and a particle. Based on information from the monitoring, an additional amount of a formulation comprising a stabilizing oligonucleotide and a particle can be administered.

Dosing is dependent on severity and responsiveness of the disease condition to be treated, with the course of treatment lasting from several days to several months, or until a cure is effected or a diminution of disease state is achieved. Persons of ordinary skill can easily determine optimum dosages, dosing methodologies and repetition rates. Optimum dosages may vary depending on the relative potency of individual compounds, and can generally be estimated based on EC50s found to be effective in in vitro and in vivo animal models.

In one embodiment, the administration of a formulation comprising a stabilizing oligonucleotide and a particle is parenteral, e.g. intravenous (e.g., as a bolus or as a diffusible infusion), intradermal, intraperitoneal, intramuscular, intrathecal, intraventricular, intracranial, subcutaneous, transmucosal, buccal, sublingual, endoscopic, rectal, oral, vaginal, topical, pulmonary, intranasal, urethral or ocular. Administration can be provided by the subject or by another person, e.g., a health care provider. The composition can be provided in measured doses or in a dispenser which delivers a metered dose. Selected modes of delivery are discussed in more detail below.

Kits

In certain aspects of the disclosure, kits are provided, comprising a container housing a formulation comprising a stabilizing oligonucleotide and a particle. In some embodiments, the formulation comprises a stabilizing oligonucleotide and a nanoparticle, e.g., lipid nanoparticle. In some embodiments, the formulation further comprises a synthetic RNA. In some embodiments, the individual components of the formulation may be provided in one container. Alternatively, it may be desirable to provide the components of the formulation separately in two or more containers, e.g., one container for stabilizing oligonucleotide, and at least another for the particle. In some embodiments, it is desirable to provide a further container for a synthetic RNA. The kit may be packaged in a number of different configurations such as one or more containers in a single box. The different components can be combined, e.g., according to instructions provided with the kit. The components can be combined according to a method described herein, e.g., to prepare and administer a pharmaceutical composition. The kit can also include a delivery device.

The present disclosure is further illustrated by the following Examples, which in no way should be construed as further limiting. The entire contents of all of the references (including literature references, issued patents, published patent applications, and co-pending patent applications) cited throughout this application are hereby expressly incorporated by reference.

EXAMPLES
Example 1. Oligonucleotide for Targeting 5′ and 3′ Ends of RNAs

Several exemplary oligonucleotide design schemes are provided herein for increasing mRNA stability. With regard to oligonucleotides targeting the 3′ end of an RNA, at least two exemplary design schemes are provided. As a first scheme, an oligo nucleotide is designed to be complementary to the 3′ end of an RNA, before the poly-A tail (FIG. 1). As a second scheme, an oligonucleotide is designed to be complementary to the 3′ end of RNA with a 5′ poly-T region that hybridizes to a poly-A tail (FIG. 1).

With regard to oligonucleotides targeting the 5′ end of an RNA, at least three exemplary design schemes are provided. For scheme one, an oligonucleotide is designed to be complementary to the 5′ end of RNA (FIG. 2). For scheme two, an oligonucleotide is designed to be complementary to the 5′ end of RNA and has a 3′ overhang to create a RNA-oligo duplex with a recessed end. In this example, the overhang is one or more C nucleotides, e.g., two Cs, which can potentially interact with a 5′ methylguanosine cap and stabilize the cap further (FIG. 2). The overhang could also potentially be another type of nucleotide, and is not limited to C. For scheme 3, an oligonucleotide is designed to include a loop region to stabilize 5′ RNA cap (FIG. 3A). FIG. 3A also shows an exemplary oligo for targeting 5′ and 3′ RNA ends. The example shows oligos that bind to a 5′ and 3′ RNA end to create a pseudo-circularized RNA (FIG. 3B).

An oligonucleotide designed as described in Example 1 may be used or evaluated for its ability to upregulate RNA by increasing mRNA stability using the methods outlined in Example 2.

Example 2: Oligos for Targeting the 5′ and 3′ End of Frataxin
Oligonucleotide Design

Oligonucleotides were designed to target the 5′ and 3′ ends of coding or non-coding mRNA. The 3′ end oligonucleotides were designed by identifying putative mRNA 3′ ends using quantitative end analysis of poly-A tails as described previously (see, e.g., Ozsolak et al. Comprehensive Polyadenylation Site Maps in Yeast and Human Reveal Pervasive Alternative Polyadenylation. Cell. Volume 143, Issue 6, 2010, Pages 1018-1029). The 5′ end oligonucleotides were designed by identifying potential 5′ start sites using Cap analysis gene expression (CAGE) as previously described (see, e.g., Cap analysis gene expression for high-throughput analysis of transcriptional starting point and identification of promoter usage. Proc Natl Acad Sci USA. 100 (26): 15776-81. 2003 Dec. 23 and Zhao, Xiaobei (2011). “Systematic Clustering of Transcription Start Site Landscapes”. PLoS ONE (Public Library of Science) 6 (8): e23409).

The sequence and structure of each oligonucleotide is shown in Tables 3-13. Table 14 provides a description of the nucleotide analogs, modifications and intranucleotide linkages used for certain oligonucleotides tested and described in the Tables. Certain oligos in Table 3 and Table 4 have two oligo names the “Oligo Name” and the “Alternative Oligo Name”, which are used interchangeably herein and are to be understood to refer to the same oligo.

TABLE 3

Oligonucleotides targeting 5′ and 3′ ends of FXN

SEQ

Alternative

ID
Oligo
Oligo
Base
Targeting
Gene

NO
Name
Name
Sequence
Region
Name
Organism
Formatted Sequence

1
Oligo48
FXN-371
TGACCCA
5′-End
FXN
human
dTs; InaGs; dAs; InaCs; dCs;

AGGGAGA

InaCs; dAs; InaAs; dGs; InaGs;

C

dGs; InaAs; dGs; InaAs; dC-Sup

2
Oligo49
FXN-372
TGGCCAC
5′-End
FXN
human
dTs; InaGs; dGs; InaCs; dCs;

TGGCCGC

InaAs; dCs; InaTs; dGs; InaGs;

A

dCs; InaCs; dGs; InaCs; dA-Sup

3
Oligo50
FXN-373
CGGCGAC
5′-End
FXN
human
dCs; InaGs; dGs; InaCs; dGs;

CCCTGGT

InaAs; dCs; InaCs; dCs; InaCs;

G

dTs; InaGs; dGs; InaTs; dG-Sup

4
Oligo51
FXN-374
CGCCCTCC
5′-End
FXN
human
dCs; InaGs; dCs; InaCs; dCs;

AGCGCTG

InaTs; dCs; InaCs; dAs; InaGs;

dCs; InaGs; dCs; InaTs; dG-Sup

5
Oligo52
FXN-375
CGCTCCG
5′-End
FXN
human
dCs; InaGs; dCs; InaTs; dCs;

CCCTCCA

InaCs; dGs; InaCs; dCs; InaCs;

G

dTs; InaCs; dCs; InaAs; dG-Sup

6
Oligo53
FXN-376
TGACCCA
5′-End
FXN
human
dTs; InaGs; dAs; InaCs; dCs;

AGGGAGA

InaCs; dAs; InaAs; dGs; InaGs;

CCC

dGs; InaAs; dGs; InaAs; dCs;

InaCs; dC-Sup

7
Oligo54
FXN-377
TGGCCAC
5′-End
FXN
human
dTs; InaGs; dGs; InaCs; dCs;

TGGCCGC

InaAs; dCs; InaTs; dGs; InaGs;

ACC

dCs; InaCs; dGs; InaCs; dAs;

InaCs; dC-Sup

8
Oligo55
FXN-378
CGGCGAC
5′-End
FXN
human
dCs; InaGs; dGs; InaCs; dGs;

CCCTGGT

InaAs; dCs; InaCs; dCs; InaCs;

GCC

dTs; InaGs; dGs; InaTs; dGs;

InaCs; dC-Sup

9
Oligo56
FXN-379
CGCCCTCC
5′-End
FXN
human
dCs; InaGs; dCs; InaCs; dCs;

AGCGCTG

InaTs; dCs; InaCs; dAs; InaGs;

CC

dCs; InaGs; dCs; InaTs; dGs;

InaCs; dC-Sup

10
Oligo57
FXN-380
CGCTCCG
5′-End
FXN
human
dCs; InaGs; dCs; InaTs; dCs;

CCCTCCA

InaCs; dGs; InaCs; dCs; InaCs;

GCC

dTs; InaCs; dCs; InaAs; dGs;

InaCs; dC-Sup

11
Oligo58
FXN-381
TGACCCA
5′-End
FXN
human
dTs; InaGs; dAs; InaCs; dCs;

AGGGAGA

InaCs; dAs; InaAs; dGs; InaGs;

CGGAAAC

dGs; InaAs; dGs; InaAs; dCs;

CAC

InaGs; dGs; dAs; dAs; dAs; dCs;

InaCs; dAs; InaC-Sup

12
Oligo59
FXN-382
TGGCCAC
5′-End
FXN
human
dTs; InaGs; dGs; InaCs; dCs;

TGGCCGC

InaAs; dCs; InaTs; dGs; InaGs;

AGGAAAC

dCs; InaCs; dGs; InaCs; dAs;

CAC

InaGs; dGs; dAs; dAs; dAs; dCs;

InaCs; dAs; InaC-Sup

13
Oligo60
FXN-383
CGGCGAC
5′-End
FXN
human
dCs; InaGs; dGs; InaCs; dGs;

CCCTGGT

InaAs; dCs; InaCs; dCs; InaCs;

GGGAAAC

dTs; InaGs; dGs; InaTs; dGs;

CTC

InaGs; dGs; dAs; dAs; dAs; dCs;

InaCs; dTs; InaC-Sup

14
Oligo61
FXN-384
CGCCCTCC
5′-End
FXN
human
dCs; InaGs; dCs; InaCs; dCs;

AGCGCTG

InaTs; dCs; InaCs; dAs; InaGs;

GGAAACC

dCs; InaGs; dCs; InaTs; dGs;

TC

InaGs; dGs; dAs; dAs; dAs; dCs;

InaCs; dTs; InaC-Sup

15
Oligo62
FXN-385
CGCTCCG
5′-End
FXN
human
dCs; InaGs; dCs; InaTs; dCs;

CCCTCCA

InaCs; dGs; InaCs; dCs; InaCs;

GCCAAAG

dTs; InaCs; dCs; InaAs; dGs;

GTC

InaCs; dCs; dAs; dAs; dAs; dGs;

InaGs; dTs; InaC-Sup

16
Oligo63
FXN-386
GGTTTTTA
3′-End
FXN
human
dGs; InaGs; dTs; InaTs; dTs;

AGGCTTT

InaTs; dTs; InaAs; dAs; InaGs;

dGs; InaCs; dTs; InaTs; dT-Sup

17
Oligo64
FXN-387
GGGGTCT
3′-End
FXN
human
dGs; InaGs; dGs; InaGs; dTs;

TGGCCTG

InaCs; dTs; InaTs; dGs; InaGs;

A

dCs; InaCs; dTs; InaGs; dA-

Sup

18
Oligo65
FXN-388
CATAATG
3′-End
FXN
human
dCs; InaAs; dTs; InaAs; dAs;

AAGCTGG

InaTs; dGs; InaAs; dAs; InaGs;

G

dCs; InaTs; dGs; InaGs; dG-Sup

19
Oligo66
FXN-389
AGGAGGC
3′-End
FXN
human
dAs; InaGs; dGs; InaAs; dGs;

AACACAT

InaGs; dCs; InaAs; dAs; InaCs;

T

dAs; InaCs; dAs; InaTs; dT-

Sup

20
Oligo67
FXN-390
ATTATTTT
3′-End
FXN
human
dAs; InaTs; dTs; InaAs; dTs;

GCTTTTT

InaTs; dTs; InaTs; dGs; InaCs;

dTs; InaTs; dTs; InaTs; dT-Sup

21
Oligo68
FXN-391
CATTTTCC
3′-End
FXN
human
dCs; InaAs; dTs; InaTs; dTs;

CTCCTGG

InaTs; dCs; InaCs; dCs; InaTs;

dCs; InaCs; dTs; InaGs; dG-Sup

22
Oligo69
FXN-392
GTAGGCT
3′-End
FXN
human
dGs; InaTs; dAs; InaGs; dGs;

ACCCTTTA

InaCs; dTs; InaAs; dCs; InaCs;

dCs; InaTs; dTs; InaTs; dA-Sup

23
Oligo70
FXN-393
GAGGCTT
3′-End
FXN
human
dGs; InaAs; dGs; InaGs; dCs;

GTTGCTTT

InaTs; dTs; InaGs; dTs; InaTs;

dGs; InaCs; dTs; InaTs; dT-Sup

24
Oligo71
FXN-394
CATGTAT
3′-End
FXN
human
dCs; InaAs; dTs; InaGs; dTs;

GATGTTA

InaAs; dTs; InaGs; dAs; InaTs;

T

dGs; InaTs; dTs; InaAs; dT-Sup

25
Oligo72
FXN-395
TTTTTGGT
3′-End
FXN
human
dTs; InaTs; dTs; InaTs; dTs;

TTTTAAG

InaGs; dGs; InaTs; dTs; InaTs;

GCTTT

dTs; InaTs; dAs; InaAs; dGs;

InaGs; dCs; InaTs; dTs; InaT-Sup

26
Oligo73
FXN-396
TTTTTGG
3′-End
FXN
human
dTs; InaTs; dTs; InaTs; dTs;

GGTCTTG

InaGs; dGs; InaGs; dGs; InaTs;

GCCTGA

dCs; InaTs; dTs; InaGs; dGs;

InaCs; dCs; InaTs; dGs; InaA-Sup

27
Oligo74
FXN-397
TTTTTCAT
3′-End
FXN
human
dTs; InaTs; dTs; InaTs; dTs;

AATGAAG

InaCs; dAs; InaTs; dAs; InaAs;

CTGGG

dTs; InaGs; dAs; InaAs; dGs;

InaCs; dTs; InaGs; dGs; InaG-Sup

28
Oligo75
FXN-398
TTTTTAGG
3′-End
FXN
human
dTs; InaTs; dTs; InaTs; dTs;

AGGCAAC

InaAs; dGs; InaGs; dAs; InaGs;

ACATT

dGs; InaCs; dAs; InaAs; dCs;

InaAs; dCs; InaAs; dTs; InaT-Sup

29
Oligo76
FXN-399
TTTTTATT
3′-End
FXN
human
dTs; InaTs; dTs; InaTs; dTs;

ATTTTGCT

InaAs; dTs; InaTs; dAs; InaTs;

TTTT

dTs; InaTs; dTs; InaGs; dCs;

InaTs; dTs; InaTs; dTs; InaT-Sup

30
Oligo77
FXN-400
TTTTTCAT
3′-End
FXN
human
dTs; InaTs; dTs; InaTs; dTs;

TTTCCCTC

InaCs; dAs; InaTs; dTs; InaTs;

CTGG

dTs; InaCs; dCs; InaCs; dTs;

InaCs; dCs; InaTs; dGs; InaG-Sup

31
Oligo78
FXN-401
TTTTTGTA
3′-End
FXN
human
dTs; InaTs; dTs; InaTs; dTs;

GGCTACC

InaGs; dTs; InaAs; dGs; InaGs;

CTTTA

dCs; InaTs; dAs; InaCs; dCs;

InaCs; dTs; InaTs; dTs; InaA-Sup

32
Oligo79
FXN-402
TTTTTGAG
3′-End
FXN
human
dTs; InaTs; dTs; InaTs; dTs;

GCTTGTT

InaGs; dAs; InaGs; dGs; InaCs;

GCTTT

dTs; InaTs; dGs; InaTs; dTs;

InaGs; dCs; InaTs; dTs; InaT-Sup

33
Oligo80
FXN-403
TTTTTCAT
3′-End
FXN
human
dTs; InaTs; dTs; InaTs; dTs;

GTATGAT

InaCs; dAs; InaTs; dGs; InaTs;

GTTAT

dAs; InaTs; dGs; InaAs; dTs;

InaGs; dTs; InaTs; dAs; InaT-Sup

TABLE 4

Other oligonucleotides targeting FXN

SEQ

Alternative

ID
Oligo
Oligo
Base
Targeting
Gene

Formatted

NO
Name
Name
Sequence
Region
Name
Organism
Sequence

34
Oligo1
FXN-324
CGGCGCC
Internal
FXN
human
dCs; InaGs; dGs; InaCs;

CGAGAGT

dGs; InaCs; dCs; InaCs;

CCACAT

dGs; InaAs; dGs;

InaAs; dGs; InaTs; dCs;

InaCs; dAs; InaCs;

dAs; InaT-Sup

35
Oligo2
FXN-325
CCAGGAG
Internal
FXN
human
dCs; InaCs; dAs; InaGs;

GCCGGCT

dGs; InaAs; dGs; InaGs;

ACTGCG

dCs; InaCs; dGs; InaGs;

dCs; InaTs; dAs;

InaCs; dTs; InaGs; dCs;

InaG-Sup

36
Oligo3
FXN-326
CTGGGCT
Internal
FXN
human
dCs; InaTs; dGs; InaGs;

GGGCTGG

dGs; InaCs; dTs; InaGs;

GTGACG

dGs; InaGs; dCs;

InaTs; dGs; InaGs; dGs;

InaTs; dGs; InaAs;

dCs; InaG-Sup

37
Oligo4
FXN-327
ACCCGGG
Internal
FXN
human
dAs; InaCs; dCs; InaCs;

TGAGGGT

dGs; InaGs; dGs; InaTs;

CTGGGC

dGs; InaAs; dGs;

InaGs; dGs; InaTs; dCs;

InaTs; dGs; InaGs;

dGs; InaC-Sup

38
Oligo5
FXN-328
CCAACTCT
Internal
FXN
human
dCs; InaCs; dAs; InaAs;

GCCGGCC

dCs; InaTs; dCs; InaTs;

GCGGG

dGs; InaCs; dCs; InaGs;

dGs; InaCs; dCs;

InaGs; dCs; InaGs; dGs;

InaG-Sup

39
Oligo6
FXN-329
ACGGCGG
Internal
FXN
human
dAs; InaCs; dGs; InaGs;

CCGCAGA

dCs; InaGs; dGs; InaCs;

GTGGGG

dCs; InaGs; dCs;

InaAs; dGs; InaAs; dGs;

InaTs; dGs; InaGs; dGs;

InaG-Sup

40
Oligo7
FXN-330
TCGATGT
Internal
FXN
human
dTs; InaCs; dGs; InaAs;

CGGTGCG

dTs; InaGs; dTs; InaCs;

CAGGCC

dGs; InaGs; dTs;

InaGs; dCs; InaGs; dCs;

InaAs; dGs; InaGs; dCs;

InaC-Sup

41
Oligo8
FXN-331
GGCGGGG
Internal
FXN
human
dGs; InaGs; dCs; InaGs;

CGTGCAG

dGs; InaGs; dGs; InaCs;

GTCGCA

dGs; InaTs; dGs;

InaCs; dAs; InaGs; dGs;

InaTs; dCs; InaGs;

dCs; InaA-Sup

42
Oligo9
FXN-332
ACGTTGG
Internal
FXN
human
dAs; InaCs; dGs; InaTs;

TTCGAACT

dTs; InaGs; dGs; InaTs;

TGCGC

dTs; InaCs; dGs;

InaAs; dAs; InaCs; dTs;

InaTs; dGs; InaCs; dGs;

InaC-Sup

43
Oligo10
FXN-333
TTCCAAAT
Internal
FXN
human
dTs; InaTs; dCs; InaCs;

CTGGTTG

dAs; InaAs; dAs; InaTs;

AGGCC

dCs; InaTs; dGs; InaGs;

dTs; InaTs; dGs; InaAs;

dGs; InaGs; dCs;

InaC-Sup

44
Oligo1l
FXN-334
AGACACT
Internal
FXN
human
dAs; InaGs; dAs; InaCs;

CTGCTTTT

dAs; InaCs; dTs; InaCs;

TGACA

dTs; InaGs; dCs; InaTs;

dTs; InaTs; dTs;

InaTs; dGs; InaAs; dCs;

InaA-Sup

45
Oligo12
FXN-335
TTTCCTCA
Internal
FXN
human
dTs; InaTs; dTs; InaCs;

AATTCATC

dCs; InaTs; dCs; InaAs;

AAAT

dAs; InaAs; dTs; InaTs;

dCs; InaAs; dTs;

InaCs; dAs; InaAs; dAs;

InaT-Sup

46
Oligo13
FXN-336
GGGTGGC
Internal
FXN
human
dGs; InaGs; dGs; InaTs;

CCAAAGT

dGs; InaGs; dCs; InaCs;

TCCAGA

dCs; InaAs; dAs;

InaAs; dGs; InaTs; dTs;

InaCs; dCs; InaAs; dGs;

InaA-Sup

47
Oligo14
FXN-337
TGGTCTC
Internal
FXN
human
dTs; InaGs; dGs; InaTs;

ATCTAGA

dCs; InaTs; dCs; InaAs;

GAGCCT

dTs; InaCs; dTs;

InaAs; dGs; InaAs; dGs;

InaAs; dGs; InaCs; dCs;

InaT-Sup

48
Oligo15
FXN-338
CTCTGCTA
Internal
FXN
human
dCs; InaTs; dCs; InaTs;

GTCTTTCA

dGs; InaCs; dTs; InaAs;

TAGG

dGs; InaTs; dCs; InaTs;

dTs; InaTs; dCs;

InaAs; dTs; InaAs; dGs;

InaG-Sup

49
Oligo16
FXN-339
GCTAAAG
Internal
FXN
human
dGs; InaCs; dTs; InaAs;

AGTCCAG

dAs; InaAs; dGs; InaAs;

CGTTTC

dGs; InaTs; dCs;

InaCs; dAs; InaGs; dCs;

InaGs; dTs; InaTs; dTs;

InaC-Sup

50
Oligo17
FXN-340
GCAAGGT
Internal
FXN
human
dGs; InaCs; dAs; InaAs;

CTTCAAA

dGs; InaGs; dTs; InaCs;

AAACTCT

dTs; InaTs; dCs;

InaAs; dAs; InaAs; dAs;

InaAs; dAs; InaCs; dTs;

InaCs; dT-Sup

51
Oligo18
FXN-341
CTCAAAC
Internal
FXN
human
dCs; InaTs; dCs; InaAs;

GTGTATG

dAs; InaAs; dCs; InaGs;

GCTTGTCT

dTs; InaGs; dTs;

InaAs; dTs; InaGs; dGs;

InaCs; dTs; InaTs; dGs;

InaTs; dCs; InaT-Sup

52
Oligo19
FXN-342
CCCAAAG
Internal
FXN
human
dCs; InaCs; dCs; InaAs;

GAGACAT

dAs; InaAs; dGs; InaGs;

CATAGTC

dAs; InaGs; dAs;

InaCs; dAs; InaTs; dCs;

InaAs; dTs; InaAs; dGs;

InaTs; dC-Sup

53
Oligo20
FXN-343
CAGTTTG
Internal
FXN
human
dCs; InaAs; dGs; InaTs;

ACAGTTA

dTs; InaTs; dGs; InaAs;

AGACACC

dCs; InaAs; dGs;

ACT

InaTs; dTs; InaAs; dAs;

InaGs; dAs; InaCs; dAs;

InaCs; dCs; InaAs; dCs;

InaT-Sup

54
Oligo21
FXN-344
ATAGGTT
Internal
FXN
human
dAs; InaTs; dAs; InaGs;

CCTAGAT

dGs; InaTs; dTs; InaCs;

CTCCACC

dCs; InaTs; dAs;

InaGs; dAs; InaTs; dCs;

InaTs; dCs; InaCs; dAs;

InaCs; dC-Sup

55
Oligo22
FXN-345
GGCGTCT
Internal
FXN
human
dGs; InaGs; dCs; InaGs;

GCTTGTT

dTs; InaCs; dTs; InaGs;

GATCAC

dCs; InaTs; dTs;

InaGs; dTs; InaTs; dGs;

InaAs; dTs; InaCs; dAs;

InaC-Sup

56
Oligo23
FXN-346
AAGATAG
Internal
FXN
human
dAs; InaAs; dGs; InaAs;

CCAGATTT

dTs; InaAs; dGs; InaCs;

GCTTGTTT

dCs; InaAs; dGs;

InaAs; dTs; InaTs; dTs;

InaGs; dCs; InaTs; dTs;

InaGs; dTs; InaTs; dT-

Sup

57
Oligo24
FXN-347
GGTCCAC
Internal
FXN
human
dGs; InaGs; dTs; InaCs;

TACATACC

dCs; InaAs; dCs; InaTs;

TGGATGG

dAs; InaCs; dAs;

AG

InaTs; dAs; InaCs; dCs;

InaTs; dGs; InaGs; dAs;

InaTs; dGs; InaGs; dAs;

InaG-Sup

58
Oligo25
FXN-348
CCCAGTC
Internal
FXN
human
dCs; InaCs; dCs; InaAs;

CAGTCAT

dGs; InaTs; dCs; InaCs;

AACGCTT

dAs; InaGs; dTs;

InaCs; dAs; InaTs; dAs;

InaAs; dCs; InaGs; dCs;

InaTs; dT-Sup

59
Oligo26
FXN-349
CGTGGGA
Internal
FXN
human
dCs; InaGs; dTs; InaGs;

GTACACC

dGs; InaGs; dAs; InaGs;

CAGTTTTT

dTs; InaAs; dCs;

InaAs; dCs; InaCs; dCs;

InaAs; dGs; InaTs; dTs;

InaTs; dTs; InaT-Sup

60
Oligo27
FXN-350
CATGGAG
Internal
FXN
human
dCs; InaAs; dTs; InaGs;

GGACACG

dGs; InaAs; dGs;

CCGT

InaGs; dGs; InaAs; dCs;

InaAs; dCs; InaGs; dCs;

InaCs; dGs; InaT-

Sup

61
Oligo28
FXN-351
GTGAGCT
Internal
FXN
human
dGs; InaTs; dGs; InaAs;

CTGCGGC

dGs; InaCs; dTs;

CAGCAGC

InaCs; dTs; InaGs; dCs;

T

InaGs; dGs; InaCs; dCs;

InaAs; dGs; InaCs; dAs;

InaGs; dCs; InaT-

Sup

62
Oligo29
FXN-352
AGTTTGG
Internal
FXN
human
dAs; InaGs; dTs; InaTs;

TTTTTAAG

dTs; InaGs; dGs;

GCTTTA

InaTs; dTs; InaTs; dTs;

InaTs; dAs; InaAs; dGs;

InaGs; dCs; InaTs; dTs;

InaTs; dA-Sup

63
Oligo30
FXN-353
TAGGCCA
Internal
FXN
human
dTs; InaAs; dGs; InaGs;

AGGAAGA

dCs; InaCs; dAs; InaAs;

CAAGTCC

dGs; InaGs; dAs;

InaAs; dGs; InaAs; dCs;

InaAs; dAs; InaGs;

dTs; InaCs; dC-Sup

64
Oligo31
FXN-354
TCAAGCA
Internal
FXN
human
dTs; InaCs; dAs; InaAs;

TCTTTTCC

dGs; InaCs; dAs; InaTs;

GGAA

dCs; InaTs; dTs;

InaTs; dTs; InaCs; dCs;

naGs; dGs; InaAs; dA-

Sup

65
Oligo32
FXN-355
TCCTTAAA
Internal
FXN
human
dTs; InaCs; dCs; InaTs;

ACGGGGC

dTs; InaAs; dAs; InaAs;

TGGGCA

dAs; InaCs; dGs; InaGs;

dGs; InaGs; dCs;

InaTs; dGs; InaGs; dGs;

InaCs; dA-Sup

66
Oligo33
FXN-356
TTGGCCT
Internal
FXN
human
dTs; InaTs; dGs; InaGs;

GATAGCT

dCs; InaCs; dTs; InaGs;

TTTAATG

dAs; InaTs; dAs;

InaGs; dCs; InaTs; dTs;

InaTs; dTs; InaAs; dAs;

InaTs; dG-Sup

67
Oligo34
FXN-357
CCTCAGCT
Internal
FXN
human
dCs; InaCs; dTs; InaCs;

GCATAAT

dAs; InaGs; dCs; InaTs;

GAAGCTG

dGs; InaCs; dAs; InaTs;

GGGTC

dAs; InaAs; dTs;

InaGs; dAs; InaAs; dGs;

InaCs; dTs; InaGs; dGs;

InaGs; dGs; InaTs;

dC-Sup

68
Oligo35
FXN-358
AACAACA
Internal
FXN
human
dAs; InaAs; dCs; InaAs;

ACAACAA

dAs; InaCs; dAs; InaAs;

CAAAAAA

dCs; InaAs; dAs;

CAGA

InaCs; dAs; InaAs; dCs;

InaAs; dAs; InaAs; dAs;

InaAs; dAs; InaCs;

dAs; InaGs; dA-Sup

69
Oligo36
FXN-359
CCTCAAA
Internal
FXN
human
dCs; InaCs; dTs; InaCs;

AGCAGGA

dAs; InaAs; dAs; InaAs;

ATAAAAA

dGs; InaCs; dAs;

AAATA

InaGs; dGs; InaAs; dAs;

InaTs; dAs; InaAs; dAs;

InaAs; dAs; InaAs;

dAs; InaAs; dTs; InaA-

Sup

70
Oligo37
FXN-360
GCTGTGA
Internal
FXN
human
dGs; InaCs; dTs; InaGs;

CACATAG

dTs; InaGs; dAs; InaCs;

CCCAACT

dAs; InaCs; dAs;

GT

InaTs; dAs; InaGs; dCs;

InaCs; dCs; InaAs; dAs;

InaCs; dTs; InaGs; dT-

Sup

71
Oligo38
FXN-361
GGAGGCA
Internal
FXN
human
dGs; InaGs; dAs; InaGs;

ACACATTC

dGs; InaCs; dAs; InaAs

TTTCTACA

dCs; InaAs; dCs;

GA

InaAs; dTs; InaTs; dCs;

InaTs; dTs; InaTs; dCs;

InaTs; dAs; InaCs; dAs;

InaGs; dA-Sup

72
Oligo39
FXN-362
CTATTAAT
Intron
FXN
human
dCs; InaTs; dAs; InaTs;

ATTACTG

dTs; InaAs; dAs; InaTs;

dAs; InaTs; dTs; InaAs;

dCs; InaTs; dG-

Sup

73
Oligo40
FXN-363
CATTATGT
Intron
FXN
human
dCs; InaAs; dTs; InaTs;

GTATGTA

dAs; InaTs; dGs; InaTs;

T

dGs; InaTs; dAs; InaTs;

dGs; InaTs; dAs; InaT-

Sup

74
Oligo41
FXN-364
TTTATCTA
Intron
FXN
human
dTs; InaTs; dTs; InaAs;

TGTTATT

dTs; InaCs; dTs; InaAs;

dTs; InaGs; dTs; InaTs;

dAs; InaTs; dT-

Sup

75
Oligo42
FXN-365
CTAATTTG
Intron
FXN
human
dCs; InaTs; dAs; InaAs;

AAGTTCT

dTs; InaTs; dTs; InaGs;

dAs; InaAs; dGs;

InaTs; dTs; InaCs; dT-

Sup

76
Oligo43
FXN-366
TTCGAACT
Exon
FXN
human
dTs; InaTs; dCs; InaGs;

TGCGCGG
Spanning

dAs; InaAs; dCs; InaTs;

dTs; InaGs; dCs;

InaGs; dCs; InaGs; dG-

Sup

77
Oligo44
FXN-367
TAGAGAG
Exon
FXN
human
dTs; InaAs; dGs; InaAs;

CCTGGGT
Spanning

dGs; InaAs; dGs; InaCs;

dCs; InaTs; dGs;

InaGs; dGs; InaT-Sup

78
Oligo45
FXN-368
ACACCAC
Exon
FXN
human
dAs; InaCs; dAs; InaCs;

TCCCAAA
Spanning

dCs; InaAs; dCs; InaTs;

G

dCs; InaCs; dCs;

InaAs; dAs; InaAs; dG-

Sup

79
Oligo46
FXN-369
AGGTCCA
Exon
FXN
human
dAs; InaGs; dGs; InaTs;

CTACATAC
Spanning

dCs; InaCs; dAs; InaCs;

dTs; InaAs; dCs;

InaAs; dTs; InaAs; dC-

Sup

80
Oligo47
FXN-370
CGTTAAC
Exon
FXN
human
dCs; InaGs; dTs; InaTs;

CTGGATG
Spanning

dAs; InaAs; dCs; InaCs;

G

dTs; InaGs; dGs;

InaAs; dTs; InaGs; dG-

Sup

81
Oligo81
FXN-404
AAAGCCT
Antisense
FXN
human
dAs; InaAs; dAs; InaGs;

TAAAAAC

dCs; InaCs; dTs; InaTs;

C

dAs; InaAs; dAs;

InaAs; dAs; InaCs; dC-

Sup

82
Oligo82
FXN-405
TCAGGCC
Antisense
FXN
human
dTs; InaCs; dAs; InaGs;

AAGACCC

dGs; InaCs; dCs; InaAs;

C

dAs; InaGs; dAs;

InaCs; dCs; InaCs; dC-

Sup

83
Oligo83
FXN-406
CCCAGCTT
Antisense
FXN
human
dCs; InaCs; dCs; InaAs;

CATTATG

dGs; InaCs; dTs; InaTs;

dCs; InaAs; dTs;

InaTs; dAs; InaTs; dG-

Sup

84
Oligo84
FXN-407
AATGTGT
Antisense
FXN
human
dAs; InaAs; dTs; InaGs;

TGCCTCCT

dTs; InaGs; dTs; InaTs;

dGs; InaCs; dCs;

InaTs; dCs; InaCs; dT-

Sup

85
Oligo85
FXN-408
AAAAAGC
Antisense
FXN
human
dAs; InaAs; dAs; InaAs;

AAAATAA

dAs; InaGs; dCs; InaAs;

T

dAs; InaAs; dAs;

InaTs; dAs; InaAs; dT-

Sup

86
Oligo86
FXN-409
CCAGGAG
Antisense
FXN
human
dCs; InaCs; dAs; InaGs;

GGAAAAT

dGs; InaAs; dGs; InaGs;

G

dGs; InaAs; dAs;

InaAs; dAs; InaTs; dG-

Sup

87
Oligo87
FXN-410
TAAAGGG
Antisense
FXN
human
dTs; InaAs; dAs; InaAs;

TAGCCTA

dGs; InaGs; dGs;

C

InaTs; dAs; InaGs; dCs;

InaCs; dTs; InaAs; dC-

Sup

88
Oligo88
FXN-411
AAAGCAA
Antisense
FXN
human
dAs; InaAs; dAs; InaGs;

CAAGCCT

dCs; InaAs; dAs; InaCs;

C

dAs; InaAs; dGs;

InaCs; dCs; InaTs; dC-

Sup

89
Oligo89
FXN-412
ATAACAT
Antisense
FXN
human
dAs; InaTs; dAs; InaAs;

CATACAT

dCs; InaAs; dTs; InaCs;

G

dAs; InaTs; dAs;

InaCs; dAs; InaTs; dG-

Sup

90
Oligo90
FXN-413
GATACTA
Antisense
FXN
human
dGs; InaAs; dTs; InaAs;

TCTTCCTC

dCs; InaTs; dAs; InaTs;

dCs; InaTs; dTs;

InaCs; dCs; InaTs; dC-

Sup

91
Oligo91
FXN-414
ATGGGGG
Antisense
FXN
human
dAs; InaTs; dGs; InaGs;

ACGGGGC

dGs; InaGs; dGs;

A

InaAs; dCs; InaGs; dGs;

InaGs; dGs; InaCs; dA-

Sup

92
Oligo92
FXN-415
GGTTGAG
Antisense
FXN
human
dGs; InaGs; dTs; InaTs;

ACTGGGT

dGs; InaAs; dGs; InaAs;

G

dCs; InaTs; dGs;

InaGs; dGs; InaTs; dG-

Sup

93
Oligo93
FXN-416
AGACTGA
Antisense
FXN
human
dAs; InaGs; dAs; InaCs;

AGAGGTG

dTs; InaGs; dAs; InaAs;

C

dGs; InaAs; dGs;

InaGs; dTs; InaGs; dC-

Sup

94
Oligo94
FXN-417
CGGGACG
Antisense
FXN
human
dCs; InaGs; dGs; InaGs;

GCTGTGT

dAs; InaCs; dGs; InaGs;

T

dCs; InaTs; dGs;

InaTs; dGs; InaTs; dT-

Sup

95
Oligo95
FXN-418
TCTGTGT
Antisense
FXN
human
dTs; InaCs; dTs; InaGs;

GGGCAGC

dTs; InaGs; dTs; InaGs;

A

dGs; InaGs; dCs;

InaAs; dGs; InaCs; dA-

Sup

96
Oligo96
FXN-419
AAAGCCT
Antisense
FXN
human
InaAs; InaAs; InaAs;

TAAAAAC

dGs; dCs; dCs; dTs; dTs;

C

dAs; dAs; dAs; dAs;

InaAs; InaCs; InaC-

Sup

97
Oligo97
FXN-420
TCAGGCC
Antisense
FXN
human
InaTs; InaCs; InaAs;

AAGACCC

dGs; dGs; dCs; dCs; dAs;

C

dAs; dGs; dAs; dCs;

InaCs; InaCs; InaC-

Sup

98
Oligo98
FXN-421
CCCAGCTT
Antisense
FXN
human
InaCs; InaCs; InaCs;

CATTATG

dAs; dGs; dCs; dTs; dTs;

dCs; dAs; dTs; dTs;

InaAs; InaTs; InaG-Sup

99
Oligo99
FXN-422
AATGTGT
Antisense
FXN
human
InaAs; InaAs; InaTs;

TGCCTCCT

dGs; dTs; dGs; dTs; dTs;

dGs; dCs; dCs; dTs;

InaCs; InaCs; InaT-Sup

100
Oligo100
FXN-423
AAAAAGC
Antisense
FXN
human
InaAs; InaAs; InaAs;

AAAATAA

dAs; dAs; dGs; dCs; dAs;

T

dAs; dAs; dAs; dTs;

InaAs; InaAs; InaT-

Sup

101
Oligo101
FXN-424
CCAGGAG
Antisense
FXN
human
InaCs; InaCs; InaAs;

GGAAAAT

dGs; dGs; dAs; dGs; dGs;

G

dGs; dAs; dAs; dAs;

InaAs; InaTs; InaG-

Sup

102
Oligo102
FXN-425
TAAAGGG
Antisense
FXN
human
InaTs; InaAs; InaAs;

TAGCCTA

dAs; dGs; dGs; dGs; dTs;

C

dAs; dGs; dCs; dCs;

InaTs; InaAs; InaC-

Sup

103
Oligo103
FXN-426
AAAGCAA
Antisense
FXN
human
InaAs; InaAs; InaAs;

CAAGCCT

dGs; dCs; dAs; dAs; dCs;

C

dAs; dAs; dGs; dCs;

InaCs; InaTs; InaC-

Sup

104
Oligo104
FXN-427
ATAACAT
Antisense
FXN
human
InaAs; InaTs; InaAs;

CATACAT

dAs; dCs; dAs; dTs; dCs;

G

dAs; dTs; dAs; dCs;

InaAs; InaTs; InaG-Sup

105
Oligo105
FXN-428
GATACTA
Antisense
FXN
human
InaGs; InaAs; InaTs;

TCTTCCTC

dAs; dCs; dTs; dAs; dTs;

dCs; dTs; dTs; dCs;

InaCs; InaTs; InaC-Sup

106
Oligo106
FXN-429
ATGGGGG
Antisense
FXN
human
InaAs; InaTs; InaGs;

ACGGGGC

dGs; dGs; dGs; dGs; dAs;

A

dCs; dGs; dGs; dGs;

InaGs; InaCs; InaA-

Sup

107
Oligo107
FXN-430
GGTTGAG
Antisense
FXN
human
InaGs; InaGs; InaTs;

ACTGGGT

dTs; dGs; dAs; dGs; dAs;

G

dCs; dTs; dGs; dGs;

InaGs; InaTs; InaG-

Sup

108
Oligo108
FXN-431
AGACTGA
Antisense
FXN
human
InaAs; InaGs; InaAs;

AGAGGTG

dCs; dTs; dGs; dAs; dAs;

C

dGs; dAs; dGs; dGs;

InaTs; InaGs; InaC-

Sup

109
Oligo109
FXN-432
CGGGACG
Antisense
FXN
human
InaCs; InaGs; InaGs;

GCTGTGT

dGs; dAs; dCs; dGs; dGs;

T

dCs; dTs; dGs; dTs;

InaGs; InaTs; InaT-

Sup

110
Oligo110
FXN-433
TCTGTGT
Antisense
FXN
human
InaTs; InaCs; InaTs;

GGGCAGC

dGs; dTs; dGs; dTs; dGs;

A

dGs; dGs; dCs; dAs;

InaGs; InaCs; InaA-

Sup

111
Oligo111
FXN-115
GAAGAAG
Antisense
FXN
human
InaGs; InaAs; InaAs;

AAGAAGA

dGs; dAs; dAs; dGs; dAs;

A

dAs; dGs; dAs; dAs;

InaGs; InaAs; InaA-

Sup

112
Oligo112
FXN-117
TTCTTCTT
Antisense
FXN
human
InaTs; InaTs; InaCs;

CTTCTTC

dTs; dTs; dCs; dTs; dTs;

dCs; dTs; dTs; dCs;

InaTs; InaTs; InaC-Sup

TABLE 5

Other targeting FXN

SEQ

ID
Oligo

Gene

Formatted

NO
Name
Base Sequence
Name
Organism
Sequence

113
324
CGGCGCCCGAGAG
FXN
human
dCs; InaGs; dGs; InaCs; dGs;

TCCACAT

InaCs; dCs; InaCs; dGs; InaAs;

dGs; InaAs; dGs; InaTs; dCs;

InaCs; dAs; InaCs; dAs; InaT-Sup

114
329
ACGGCGGCCGCAG
FXN
human
dAs; InaCs; dGs; InaGs; dCs;

AGTGGGG

InaGs; dGs; InaCs; dCs; InaGs;

dCs; InaAs; dGs; InaAs; dGs;

InaTs; dGs; InaGs; dGs; InaG-Sup

115
359
CCTCAAAAGCAGGA
FXN
human
dCs; InaCs; dTs; InaCs; dAs;

ATAAAAAAAATA

InaAs; dAs; InaAs; dGs; InaCs;

dAs; InaGs; dGs; InaAs; dAs; InaTs;

dAs; InaAs; dAs; InaAs; dAs;

InaAs; dAs; InaAs; dTs; InaA-Sup

116
414
ATGGGGGACGGGG
FXN
human
dAs; InaTs; dGs; InaGs; dGs;

CA

InaGs; dGs; InaAs; dCs; InaGs;

dGs; InaGs; dGs; InaCs; dA-Sup

117
415
GGTTGAGACTGGG
FXN
human
dGs; InaGs; dTs; InaTs; dGs;

TG

InaAs; dGs; InaAs; dCs; InaTs;

dGs; InaGs; dGs; InaTs; dG-Sup

118
429
ATGGGGGACGGGG
FXN
human
dAs; InaTs; dGs; InaGs; dGs;

CA

InaGs; dGs; InaAs; dCs; InaGs;

dGs; InaGs; dGs; InaCs; dA-Sup

Several additional oligonucleotides were designed to target the 5′ end of an RNA, the 3′ end of an RNA, or target both the 5′ end and 3′ end of an RNA (“bridging oligos”). These oligos are shown in Table 6.

TABLE 6

Oligonucleotides designed to target 5′ and 3′ ends of RNAs

SEQ
Oligo

Gene
Target

ID NO
Name
Base Sequence
Name
Region
Organism
Formatted Sequence

119
FXN-437
TGACCCAAGGGAGACTT
FXN
5′ and 3′
human
dTs; InaGs; dAs; InaCs; dCs;

m02
TTTGGTTTTTAAGGCTTT

InaCs; dAs; InaAs; dGs; InaGs;

dGs; InaAs; dGs; InaAs;

dCs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaGs; dTs; InaTs;

dTs; InaTs; dTs; InaAs; dAs;

InaGs; dGs; InaCs; dTs;

InaTs; dT-Sup

120
FXN-438
TGGCCACTGGCCGCATT
FXN
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
TTTGGTTTTTAAGGCTTT

InaAs; dCs; InaTs; dGs; InaGs;

dCs; InaCs; dGs; InaCs;

dAs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaGs; dTs; InaTs;

dTs; InaTs; dTs; InaAs; dAs;

InaGs; dGs; InaCs; dTs; InaTs;

dT-Sup

121
FXN-439
CGGCGACCCCTGGTGTT
FXN
5′ and 3′
human
dCs; InaGs; dGs; InaCs; dGs;

m02
TTTGGTTTTTAAGGCTTT

InaAs; dCs; InaCs; dCs; InaCs;

dTs; InaGs; dGs; InaTs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaGs; dTs; InaTs;

dTs; InaTs; dTs; InaAs; dAs;

InaGs; dGs; InaCs; dTs; InaTs;

dT-Sup

122
FXN-440
CGCCCTCCAGCGCTGTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTGGTTTTTAAGGCTTT

InaTs; dCs; InaCs; dAs; InaGs;

dCs; InaGs; dCs; InaTs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dGs; InaGs; dTs; InaTs;

dTs; InaTs; dTs; InaAs; dAs;

InaGs; dGs; InaCs; dTs; InaTs;

dT-Sup

123
FXN-441
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs; dCs;

m02
TTGGTTTTTAAGGCTTT

InaCs; dGs; InaCs; dCs; InaCs;

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dGs; InaGs; dTs; InaTs;

dTs; InaTs; dTs; InaAs; dAs;

InaGs; dGs; InaCs; dTs; InaTs;

dT-Sup

124
FXN-442
TGACCCAAGGGAGACTT
FXN
5′ and 3′
human
dTs; InaGs; dAs; InaCs; dCs;

m02
TTTGGGGTCTTGGCCTG

InaCs; dAs; InaAs; dGs; InaGs;

A

dGs; InaAs; dGs; InaAs;

dCs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaGs; dGs; InaGs;

dTs; InaCs; dTs; InaTs; dGs;

InaGs; dCs; InaCs; dTs; InaGs;

dA-Sup

125
FXN-443
TGGCCACTGGCCGCATT
FXN
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
TTTGGGGTCTTGGCCTG

InaAs; dCs; InaTs; dGs; InaGs;

A

dCs; InaCs; dGs; InaCs;

dAs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaGs; dGs; InaGs;

dTs; InaCs; dTs; InaTs; dGs;

InaGs; dCs; InaCs; dTs; InaGs;

dA-Sup

126
FXN-444
CGGCGACCCCTGGTGTT
FXN
5′ and 3′
human
dCs; InaGs; dGs; InaCs; dGs;

m02
TTTGGGGTCTTGGCCTG

InaAs; dCs; InaCs; dCs; InaCs;

A

dTs; InaGs; dGs; InaTs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaGs; dGs; InaGs;

dTs; InaCs; dTs; InaTs; dGs;

InaGs; dCs; InaCs; dTs; InaGs;

dA-Sup

127
FXN-445
CGCCCTCCAGCGCTGTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTGGGGTCTTGGCCTG

InaTs; dCs; InaCs; dAs; InaGs;

A

dCs; InaGs; dCs; InaTs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

; dGs; InaGs; dGs; InaGs;

dTs; InaCs; dTs; InaTs; dGs;

InaGs; dCs; InaCs; dTs; InaGs;

dA-Sup

128
FXN-446
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs; dCs;

m02
TTGGGGTCTTGGCCTGA

InaCs; dGs; InaCs; dCs; InaCs;

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dGs; InaGs; dGs; InaGs;

dTs; InaCs; dTs; InaTs; dGs;

InaGs; dCs; InaCs; dTs; InaGs;

dA-Sup

129
FXN-447
TGACCCAAGGGAGACTT
FXN
5′ and 3′
human
dTs; InaGs; dAs; InaCs; dCs;

m02
TTTCATAATGAAGCTGG

InaCs; dAs; InaAs; dGs; InaGs;

G

dGs; InaAs; dGs; InaAs;

dCs; InaTs; dTs; InaTs; dTs;

InaTs; dCs; InaAs; dTs; InaAs;

dAs; InaTs; dGs; InaAs;

dAs; InaGs; dCs; InaTs; dGs;

InaGs; dG-Sup

130
FXN-448
TGGCCACTGGCCGCATT
FXN
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
TTTCATAATGAAGCTGG

InaAs; dCs; InaTs; dGs; InaGs;

G

dCs; InaCs; dGs; InaCs;

dAs; InaTs; dTs; InaTs; dTs;

InaTs; dCs; InaAs; dTs; InaAs;

dAs; InaTs; dGs; InaAs;

dAs; InaGs; dCs; InaTs; dGs;

InaGs; dG-Sup

131
FXN-449
CGGCGACCCCTGGTGTT
FXN
5′ and 3′
human
dCs; InaGs; dGs; InaCs; dGs;

m02
TTTCATAATGAAGCTGG

InaAs; dCs; InaCs; dCs; InaCs;

G

dTs; InaGs; dGs; InaTs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dCs; InaAs; dTs; InaAs;

dAs; InaTs; dGs; InaAs; dAs;

InaGs; dCs; InaTs; dGs; InaGs;

dG-Sup

132
FXN-450
CGCCCTCCAGCGCTGTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTCATAATGAAGCTGG

InaTs; dCs; InaCs; dAs; InaGs;

G

dCs; InaGs; dCs; InaTs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dCs; InaAs; dTs; InaAs; dAs;

InaTs; dGs; InaAs; dAs;

InaGs; dCs; InaTs; dGs; InaGs;

dG-Sup

133
FXN-451
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs; dCs;

m02
TTCATAATGAAGCTGGG

InaCs; dGs; InaCs; dCs; InaCs;

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dCs; InaAs; dTs; InaAs;

dAs; InaTs; dGs; InaAs; dAs;

InaGs; dCs; InaTs; dGs; InaGs;

dG-Sup

134
FXN-452
TGACCCAAGGGAGACTT
FXN
5′ and 3′
human
dTs; InaGs; dAs; InaCs; dCs;

m02
TTTAGGAGGCAACACAT

InaCs; dAs; InaAs; dGs; InaGs;

T

dGs; InaAs; dGs; InaAs;

dCs; InaTs; dTs; InaTs; dTs;

InaTs; dAs; InaGs; dGs; InaAs;

dGs; InaGs; dCs; InaAs;

dAs; InaCs; dAs; InaCs; dAs;

InaTs; dT-Sup

135
FXN-453
TGGCCACTGGCCGCATT
FXN
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
TTTAGGAGGCAACACAT

InaAs; dCs; InaTs; dGs; InaGs;

T

dCs; InaCs; dGs; InaCs;

dAs; InaTs; dTs; InaTs; dTs;

InaTs; dAs; InaGs; dGs; InaAs;

dGs; InaGs; dCs; InaAs;

dAs; InaCs; dAs; InaCs; dAs;

InaTs; dT-Sup

136
FXN-454
CGGCGACCCCTGGTGTT
FXN
5′ and 3′
human
dCs; InaGs; dGs; InaCs; dGs;

m02
TTTAGGAGGCAACACAT

InaAs; dCs; InaCs; dCs; InaCs;

T

dTs; InaGs; dGs; InaTs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dAs; InaGs; dGs; InaAs;

dGs; InaGs; dCs; InaAs; dAs;

InaCs; dAs; InaCs; dAs;

InaTs; dT-Sup

137
FXN-455
CGCCCTCCAGCGCTGTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTAGGAGGCAACACAT

InaTs; dCs; InaCs; dAs; InaGs;

T

dCs; InaGs; dCs; InaTs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dAs; InaGs; dGs; InaAs;

dGs; InaGs; dCs; InaAs; dAs;

InaCs; dAs; InaCs; dAs; InaTs;

dT-Sup

138
FXN-456
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs; dCs;

m02
TTAGGAGGCAACACATT

InaCs; dGs; InaCs; dCs; InaCs;

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dAs; InaGs; dGs; InaAs;

dGs; InaGs; dCs; InaAs; dAs;

InaCs; dAs; InaCs; dAs; InaTs;

dT-Sup

139
FXN-457
TGACCCAAGGGAGACTT
FXN
5′ and 3′
human
dTs; InaGs; dAs; InaCs; dCs;

m02
TTTATTATTTTGCTTTTT

InaCs; dAs; InaAs; dGs; InaGs;

dGs; InaAs; dGs; InaAs;

dCs; InaTs; dTs; InaTs; dTs;

InaTs; dAs; InaTs; dTs; InaAs;

dTs; InaTs; dTs; InaTs; dGs;

InaCs; dTs; InaTs; dTs; InaTs;

dT-Sup

140
FXN-458
TGGCCACTGGCCGCATT
FXN
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
TTTATTATTTTGCTTTTT

InaAs; dCs; InaTs; dGs; InaGs;

dCs; InaCs; dGs; InaCs;

dAs; InaTs; dTs; InaTs; dTs;

InaTs; dAs; InaTs; dTs; InaAs;

dTs; InaTs; dTs; InaTs; dGs;

InaCs; dTs; InaTs; dTs; InaTs;

dT-Sup

141
FXN-459
CGGCGACCCCTGGTGTT
FXN
5′ and 3′
human
dCs; InaGs; dGs; InaCs; dGs;

m02
TTTATTATTTTGCTTTTT

InaAs; dCs; InaCs; dCs; InaCs;

dTs; InaGs; dGs; InaTs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dAs; InaTs; dTs; InaAs;

dTs; InaTs; dTs; InaTs; dGs;

InaCs; dTs; InaTs; dTs; InaTs;

dT-Sup

142
FXN-460
CGCCCTCCAGCGCTGTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTATTATTTTGCTTTTT

InaTs; dCs; InaCs; dAs; InaGs;

dCs; InaGs; dCs; InaTs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dAs; InaTs; dTs; InaAs;

dTs; InaTs; dTs; InaTs; dGs;

InaCs; dTs; InaTs; dTs; InaTs;

dT-Sup

143
FXN-461
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs; dCs;

m02
TTATTATTTTGCTTTTT

InaCs; dGs; InaCs; dCs; InaCs;

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dAs; InaTs; dTs; InaAs;

dTs; InaTs; dTs; InaTs; dGs;

InaCs; dTs; InaTs; dTs; InaTs;

dT-Sup

144
FXN-462
TGACCCAAGGGAGACTT
FXN
5′ and 3′
human
dTs; InaGs; dAs; InaCs; dCs;

m02
TTTCATTTTCCCTCCTGG

InaCs; dAs; InaAs; dGs; InaGs;

dGs; InaAs; dGs; InaAs;

dCs; InaTs; dTs; InaTs; dTs;

InaTs; dCs; InaAs; dTs; InaTs;

dTs; InaTs; dCs; InaCs; dCs;

InaTs; dCs; InaCs; dTs; InaGs;

dG-Sup

145
FXN-463
TGGCCACTGGCCGCATT
FXN
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
TTTCATTTTCCCTCCTGG

InaAs; dCs; InaTs; dGs; InaGs;

dCs; InaCs; dGs; InaCs;

dAs; InaTs; dTs; InaTs; dTs;

InaTs; dCs; InaAs; dTs; InaTs;

dTs; InaTs; dCs; InaCs; dCs;

InaTs; dCs; InaCs; dTs; InaGs;

dG-Sup

146
FXN-464
CGGCGACCCCTGGTGTT
FXN
5′ and 3′
human
dCs; InaGs; dGs; InaCs; dGs;

m02
TTTCATTTTCCCTCCTGG

InaAs; dCs; InaCs; dCs; InaCs;

dTs; InaGs; dGs; InaTs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dCs; InaAs; dTs; InaTs;

dTs; InaTs; dCs; InaCs; dCs;

InaTs; dCs; InaCs; dTs; InaGs;

dG-Sup

147
FXN-465
CGCCCTCCAGCGCTGTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTCATTTTCCCTCCTGG

InaTs; dCs; InaCs; dAs; InaGs;

dCs; InaGs; dCs; InaTs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dCs; InaAs; dTs; InaTs;

dTs; InaTs; dCs; InaCs; dCs;

InaTs; dCs; InaCs; dTs; InaGs;

dG-Sup

148
FXN-466
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs; dCs;

m02
TTCATTTTCCCTCCTGG

InaCs; dGs; InaCs; dCs; InaCs;

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dCs; InaAs; dTs; InaTs;

dTs; InaTs; dCs; InaCs; dCs;

InaTs; dCs; InaCs; dTs; InaGs;

dG-Sup

149
FXN-467
TGACCCAAGGGAGACTT
FXN
5′ and 3′
human
dTs; InaGs; dAs; InaCs; dCs;

m02
TTTGTAGGCTACCCTTTA

InaCs; dAs; InaAs; dGs; InaGs;

dGs; InaAs; dGs; InaAs;

dCs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaTs; dAs; InaGs;

dGs; InaCs; dTs; InaAs; dCs;

InaCs; dCs; InaTs; dTs; InaTs;

dA-Sup

150
FXN-468
TGGCCACTGGCCGCATT
FXN
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
TTTGTAGGCTACCCTTTA

InaAs; dCs; InaTs; dGs; InaGs;

dCs; InaCs; dGs; InaCs;

dAs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaTs; dAs; InaGs;

dGs; InaCs; dTs; InaAs; dCs;

InaCs; dCs; InaTs; dTs; InaTs;

dA-Sup

151
FXN-469
CGGCGACCCCTGGTGTT
FXN
5′ and 3′
human
dCs; InaGs; dGs; InaCs; dGs;

m02
TTTGTAGGCTACCCTTTA

InaAs; dCs; InaCs; dCs; InaCs;

dTs; InaGs; dGs; InaTs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaTs; dAs; InaGs;

dGs; InaCs; dTs; InaAs; dCs;

InaCs; dCs; InaTs; dTs; InaTs;

s; dA-Sup

152
FXN-470
CGCCCTCCAGCGCTGTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTGTAGGCTACCCTTTA

InaTs; dCs; InaCs; dAs; InaG

s; dCs; InaGs; dCs; InaTs; dG

InaTs; dTs; InaTs; dTs; InaTs;

dGs; InaTs; dAs; InaGs;

dGs; InaCs; dTs; InaAs; dCs;

InaCs; dCs; InaTs; dTs; InaTs;

dA-Sup

153
FXN-471
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs; dCs;

m02
TTGTAGGCTACCCTTTA

InaCs; dGs; InaCs; dCs; InaCs;

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dGs; InaTs; dAs; InaGs;

dGs; InaCs; dTs; InaAs; dCs;

InaCs; dCs; InaTs; dTs; InaTs;

dA-Sup

154
FXN-472
TGACCCAAGGGAGACTT
FXN
5′ and 3′
human
dTs; InaGs; dAs; InaCs; dCs;

m02
TTTGAGGCTTGTTGCTTT

InaCs; dAs; InaAs; dGs; InaGs;

dGs; InaAs; dGs; InaAs;

dCs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaAs; dGs; InaGs;

dCs; InaTs; dTs; InaGs; dTs;

InaTs; dGs; InaCs; dTs; InaTs;

dT-Sup

155
FXN-473
TGGCCACTGGCCGCATT
FXN
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
TTTGAGGCTTGTTGCTTT

InaAs; dCs; InaTs; dGs; InaGs;

dCs; InaCs; dGs; InaCs;

dAs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaAs; dGs; InaGs;

dCs; InaTs; dTs; InaGs; dTs;

InaTs; dGs; InaCs; dTs;

InaTs; dT-Sup

156
FXN-474
CGGCGACCCCTGGTGTT
FXN
5′ and 3′
human
dCs; InaGs; dGs; InaCs; dGs;

m02
TTTGAGGCTTGTTGCTTT

InaAs; dCs; InaCs; dCs; InaCs;

dTs; InaGs; dGs; InaTs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaAs; dGs; InaGs;

dCs; InaTs; dTs; InaGs; dTs;

InaTs; dGs; InaCs; dTs; InaTs;

dT-Sup

157
FXN-475
CGCCCTCCAGCGCTGTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTGAGGCTTGTTGCTTT

InaTs; dCs; InaCs; dAs; InaGs;

dCs; InaGs; dCs; InaTs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dGs; InaAs; dGs; InaGs;

dCs; InaTs; dTs; InaGs; dTs;

InaTs; dGs; InaCs; dTs; InaTs;

dT-Sup

158
FXN-476
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs; dCs;

m02
TTGAGGCTTGTTGCTTT

InaCs; dGs; InaCs; dCs; InaCs;

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

; dGs; InaAs; dGs; InaGs;

dCs; InaTs; dTs; InaGs; dTs;

InaTs; dGs; InaCs; dTs; InaTs;

dT-Sup

159
FXN-477
TGACCCAAGGGAGACTT
FXN
5′ and 3′
human
dTs; InaGs; dAs; InaCs; dCs;

m02
TTTCATGTATGATGTTAT

InaCs; dAs; InaAs; dGs; InaGs;

dGs; InaAs; dGs; InaAs;

dCs; InaTs; dTs; InaTs; dTs;

InaTs; dCs; InaAs; dTs; InaGs;

dTs; InaAs; dTs; InaGs; dAs;

InaTs; dGs; InaTs; dTs; InaAs;

dT-Sup

160
FXN-478
TGGCCACTGGCCGCATT
FXN
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
TTTCATGTATGATGTTAT

InaAs; dCs; InaTs; dGs; InaGs;

dCs; InaCs; dGs; InaCs;

dAs; InaTs; dTs; InaTs; dTs;

InaTs; dCs; InaAs; dTs; InaGs;

dTs; InaAs; dTs; InaGs; dAs;

InaTs; dGs; InaTs; dTs; InaAs;

dT-Sup

161
FXN-479
CGGCGACCCCTGGTGTT
FXN
5′ and 3′
human
dCs; InaGs; dGs; InaCs; dGs;

m02
TTTCATGTATGATGTTAT

InaAs; dCs; InaCs; dCs; InaCs;

dTs; InaGs; dGs; InaTs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dCs; InaAs; dTs; InaGs;

dTs; InaAs; dTs; InaGs; dAs;

InaTs; dGs; InaTs; dTs; InaAs;

dT-Sup

162
FXN-480
CGCCCTCCAGCGCTGTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTCATGTATGATGTTAT

InaTs; dCs; InaCs; dAs; InaGs;

dCs; InaGs; dCs; InaTs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dCs; InaAs; dTs; InaGs;

dTs; InaAs; dTs; InaGs; dAs;

InaTs; dGs; InaTs; dTs; InaAs;

dT-Sup

163
FXN-481
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs; dCs;

m02
TTCATGTATGATGTTAT

InaCs; dGs; InaCs; dCs; InaCs;

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dCs; InaAs; dTs; InaGs; dTs;

InaAs; dTs; InaGs; dAs;

InaTs; dGs; InaTs; dTs; InaAs;

dT-Sup

164
FXN-482
CGCCCTCCAGTTTTTGGT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTTAAG

InaTs; dCs; InaCs; dAs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

naGs; dGs; InaTs; dTs; InaTs;

dTs; InaTs; dAs; InaAs;

dG-Sup

165
FXN-483
CGCCCTCCAGTTTTTGG
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
GGTCTTGG

InaTs; dCs; InaCs; dAs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaGs; dGs; InaGs; dGs;

InaTs; dCs; InaTs; dTs; InaGs;

dG-Sup

166
FXN-484
CGCCCTCCAGTTTTTCAT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
AATGAAG

InaTs; dCs; InaCs; dAs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaCs; dAs; InaTs; dAs; InaAs;

dTs; InaGs; dAs; InaAs;

dG-Sup

167
FXN-485
CGCCCTCCAGTTTTTAG
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
GAGGCAAC

InaTs; dCs; InaCs; dAs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dGs; InaGs; dAs; InaGs;

dGs; InaCs; dAs; InaAs;

dC-Sup

168
FXN-486
CGCCCTCCAGTTTTTATT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
ATTTTGC

InaTs; dCs; InaCs; dAs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dTs; InaTs; dAs; InaTs;

dTs; InaTs; dTs; InaGs;

dC-Sup

169
FXN-487
CGCCCTCCAGTTTTTCAT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTCCCT

InaTs; dCs; InaCs; dAs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaCs; dAs; InaTs; dTs; InaTs;

dTs; InaCs; dCs; InaCs;

dT-Sup

170
FXN-488
CGCCCTCCAGTTTTTGTA
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
GGCTACC

InaTs; dCs; InaCs; dAs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaGs; dTs; InaAs; dGs; InaGs;

dCs; InaTs; dAs; InaCs;

dC-Sup

171
FXN-489
CGCCCTCCAGTTTTTGA
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
GGCTTGTT

InaTs; dCs; InaCs; dAs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaGs; dAs; InaGs; dGs; InaCs;

dTs; InaTs; dGs; InaTs;

dT-Sup

172
FXN-490
CGCCCTCCAGTTTTTCAT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
GTATGAT

InaTs; dCs; InaCs; dAs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaCs; dAs; InaTs; dGs; InaTs;

dAs; InaTs; dGs; InaAs;

dT-Sup

173
FXN-491
TGACCCAAGGGAGACTT
FXN
5′ and 3′
human
dTs; InaGs; dAs; InaCs; dCs;

m02
TTTTTTTTTT

InaCs; dAs; InaAs; dGs; InaGs;

dGs; InaAs; dGs; InaAs;

dCs; InaTs; dTs; InaTs; dTs;

InaTs; dTs; InaTs; dTs; InaTs;

dTs; InaTs; dT-Sup

174
FXN-492
TGGCCACTGGCCGCATT
FXN
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
TTTTTTTTTT

InaAs; dCs; InaTs; dGs; InaGs;

dCs; InaCs; dGs; InaCs;

dAs; InaTs; dTs; InaTs; dTs;

InaTs; dTs; InaTs; dTs; InaTs;

dTs; InaTs; dT-Sup

175
FXN-493
CGGCGACCCCTGGTGTT
FXN
5′ and 3′
human
dCs; InaGs; dGs; InaCs; dGs;

m02
TTTTTTTTTT

InaAs; dCs; InaCs; dCs; InaCs;

dTs; InaGs; dGs; InaTs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dTs; InaTs; dTs; InaTs;

dTs; InaTs; dT-Sup

176
FXN-494
CGCCCTCCAGCGCTGTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs; dCs;

m02
TTTTTTTTTT

InaTs; dCs; InaCs; dAs; InaGs;

dCs; InaGs; dCs; InaTs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dTs; InaTs; dTs; InaTs;

dTs; InaTs; dT-Sup

177
FXN-495
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs; dCs;

m02
TTTTTTTTT

InaCs; dGs; InaCs; dCs; InaCs;

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dTs; InaTs; dTs; InaTs;

dTs; InaTs; dTs; InaTs;

dTs; InaTs; dT-Sup

178
FXN-496
AAAATAAACAACAAC
FXN
UTR
human
dAs; InaAs; dAs; InaAs; dTs;

m02

InaAs; dAs; InaAs; dCs; InaAs;

dAs; InaCs; dAs; InaAs;

dC-Sup

179
FXN-497
AGGAATAAAAAAAATA
FXN
UTR
human
dAs; InaGs; dGs; InaAs; dAs;

m02

InaTs; dAs; InaAs; dAs; InaAs;

dAs; InaAs; dAs; InaAs;

dTs; InaA-Sup

180
FXN-498
TCAAAAGCAGGAATA
FXN
UTR
human
dTs; InaCs; dAs; InaAs; dAs;

m02

InaAs; dGs; InaCs; dAs; InaGs;

dGs; InaAs; dAs; InaTs;

dA-Sup

181
FXN-499
ACTGTCCTCAAAAGC
FXN
UTR
human
dAs; InaCs; dTs; InaGs; dTs;

m02

InaCs; dCs; InaTs; dCs; InaAs;

dAs; InaAs; dAs; InaGs;

dC-Sup

182
FXN-500
AGCCCAACTGTCCTC
FXN
UTR
human
dAs; InaGs; dCs; InaCs; dCs;

m02

InaAs; dAs; InaCs; dTs; InaGs;

dTs; InaCs; dCs; InaTs;

dC-Sup

183
FXN-501
TGACACATAGCCCAA
FXN
UTR
human
dTs; InaGs; dAs; InaCs; dAs;

m02

InaCs; dAs; InaTs; dAs; InaGs;

dCs; InaCs; dCs; InaAs;

dA-Sup

184
FXN-502
GAGCTGTGACACATA
FXN
UTR
human
dGs; InaAs; dGs; InaCs; dTs;

m02

InaGs; dTs; InaGs; dAs; InaCs;

dAs; InaCs; dAs; InaTs;

dA-Sup

185
FXN-503
TCTGGGCCTGGGCTG
FXN
UTR/internal
human
dTs; InaCs; dTs; InaGs; dGs;

m02

InaGs; dCs; InaCs; dTs; InaGs;

dGs; InaGs; dCs; InaTs;

dG-Sup

186
FXN-504
GGTGAGGGTCTGGGC
FXN
UTR/internal
human
dGs; InaGs; dTs; InaGs; dAs;

m02

InaGs; dGs; InaGs; dTs; InaCs;

dTs; InaGs; dGs; InaGs;

dC-Sup

187
FXN-505
GGGACCCGGGTGAGG
FXN
UTR/internal
human
dGs; InaGs; dGs; InaAs; dCs;

m02

InaCs; dCs; InaGs; dGs; InaGs;

dTs; InaGs; dAs; InaGs;

dG-Sup

188
FXN-506
CCGGCCGCGGGACCC
FXN
UTR/internal
human
dCs; InaCs; dGs; InaGs; dCs;

m02

InaCs; dGs; InaCs; dGs; InaGs;

dGs; InaAs; dCs; InaCs;

dC-Sup

189
FXN-507
CAACTCTGCCGGCCG
FXN
UTR/internal
human
dCs; InaAs; dAs; InaCs; dTs;

m02

InaCs; dTs; InaGs; dCs; InaCs;

dGs; InaGs; dCs; InaCs;

dG-Sup

190
FXN-508
AGTGGGGCCAACTCT
FXN
UTR/internal
human
dAs; InaGs; dTs; InaGs; dGs;

m02

InaGs; dGs; InaCs; dCs; InaAs;

dAs; InaCs; dTs; InaCs;

dT-Sup

191
FXN-509
GGCCGCAGAGTGGGG
FXN
UTR/internal
human
dGs; InaGs; dCs; InaCs; dGs;

m02

InaCs; dAs; InaGs; dAs; InaGs;

dTs; InaGs; dGs; InaGs;

dG-Sup

192
FXN-510
GCCACGGCGGCCGCA
FXN
UTR/internal
human
dGs; InaCs; dCs; InaAs; dCs;

m02

InaGs; dGs; InaCs; dGs; InaGs;

dCs; InaCs; dGs; InaCs;

dA-Sup

193
FXN-511
GTGCGCAGGCCACGG
FXN
UTR/internal
human
dGs; InaTs; dGs; InaCs; dGs;

m02

InaCs; dAs; InaGs; dGs; InaCs;

dCs; InaAs; dCs; InaGs;

dG-Sup

194
FXN-512
GGGGGACGGGGCAGG
FXN
intron
human
dGs; InaGs; dGs; InaGs; dGs;

m02

InaAs; dCs; InaGs; dGs; InaGs;

dGs; InaCs; dAs; InaGs;

dG-Sup

195
FXN-513
GGGACGGGGCAGGTT
FXN
intron
human
dGs; InaGs; dGs; InaAs; dCs;

m02

InaGs; dGs; InaGs; dGs; InaCs;

dAs; InaGs; dGs; InaTs;

dT-Sup

196
FXN-514
GACGGGGCAGGTTGA
FXN
intron
human
dGs; InaAs; dCs; InaGs; dGs;

m02

InaGs; dGs; InaCs; dAs; InaGs;

dGs; InaTs; dTs; InaGs;

dA-Sup

197
FXN-515
CGGGGCAGGTTGAGA
FXN
intron
human
dCs; InaGs; dGs; InaGs; dGs;

m02

InaCs; dAs; InaGs; dGs; InaTs;

dTs; InaGs; dAs; InaGs;

dA-Sup

198
FXN-516
GGGCAGGTTGAGACT
FXN
intron
human
dGs; InaGs; dGs; InaCs; dAs;

m02

InaGs; dGs; InaTs; dTs; InaGs;

dAs; InaGs; dAs; InaCs;

dT-Sup

199
FXN-517
GCAGGTTGAGACTGG
FXN
intron
human
dGs; InaCs; dAs; InaGs; dGs;

m02

InaTs; dTs; InaGs; dAs; InaGs;

dAs; InaCs; dTs; InaGs;

dG-Sup

200
FXN-518
AGGTTGAGACTGGGT
FXN
intron
human
dAs; InaGs; dGs; InaTs; dTs;

m02

InaGs; dAs; InaGs; dAs; InaCs;

dTs; In aGs; dGs; InaGs;

dT-Sup

201
FXN-519
GGAAAAATTCCAGGA
FXN
Antisense/
human
dGs; InaGs; dAs; InaAs; dAs;

m02

UTR

InaAs; dAs; InaTs; dTs; InaCs;

dCs; InaAs; dGs; InaGs;

dA-Sup

202
FXN-520
AATTCCAGGAGGGAA
FXN
Antisense/
human
dAs; InaAs; dTs; InaTs; dCs;

m02

UTR

InaCs; dAs; InaGs; dGs; InaAs;

dGs; InaGs; dGs; InaAs;

dA-Sup

203
FXN-521
GAGGGAAAATGAATT
FXN
Antisense/
human
dGs; InaAs; dGs; InaGs; dGs;

m02

UTR

InaAs; dAs; InaAs; dAs; InaTs;

dGs; InaAs; dAs; InaTs;

dT-Sup

204
FXN-522
GAAAATGAATTGTCTTC
FXN
Antisense/
human
dGs; InaAs; dAs; InaAs; dAs;

m02

UTR

InaTs; dGs; InaAs; dAs; InaTs;

dTs; InaGs; dTs; InaCs;

dTs; InaTs; dC-Sup

205
FXN-512
GGGGGACGGGGCAGG
FXN
intron
human
InaGs; InaGs; InaGs; dGs;

m08

dGs; dAs; dCs; dGs; dGs; dGs;

dGs; dCs; InaAs; InaGs; InaG-

Sup

206
FXN-513
GGGACGGGGCAGGTT
FXN
intron
human
InaGs; InaGs; InaGs; dAs;

m08

dCs; dGs; dGs; dGs; dGs; dCs;

dAs; dGs; InaGs; InaTs; InaT-

Sup

207
FXN-514
GACGGGGCAGGTTGA
FXN
intron
human
InaGs; InaAs; InaCs; dGs;

m08

dGs; dGs; dGs; dCs; dAs; dGs;

dGs; dTs; InaTs; InaGs; InaA-

Sup

208
FXN-515
CGGGGCAGGTTGAGA
FXN
intron
human
InaCs; InaGs; InaGs; dGs;

m08

dGs; dCs; dAs; dGs; dGs; dTs;

dTs; dGs; InaAs; InaGs; InaA-

Sup

209
FXN-516
GGGCAGGTTGAGACT
FXN
intron
human
InaGs; InaGs; InaGs; dCs;

m08

dAs; dGs; dGs; dTs; dTs; dGs;

dAs; dGs; InaAs; InaCs; InaT-

Sup

210
FXN-517
GCAGGTTGAGACTGG
FXN
intron
human
InaGs; InaCs; InaAs; dGs;

m08

dGs; dTs; dTs; dGs; dAs; dGs;

dAs; dCs; InaTs; InaGs; InaG-

Sup

211
FXN-518
AGGTTGAGACTGGGT
FXN
intron
human
InaAs; InaGs; InaGs; dTs;

m08

dTs; dGs; dAs; dGs; dAs; dCs;

dTs; dGs; InaGs; InaGs; InaT-

Sup

212
FXN-519
GGAAAAATTCCAGGA
FXN
Antisense/
human
InaGs; InaGs; InaAs; dAs;

m08

UTR

dAs; dAs; dAs; dTs; dTs; dCs;

dCs; dAs; InaGs; InaGs; InaA-

Sup

213
FXN-520
AATTCCAGGAGGGAA
FXN
Antisense/
human
InaAs; InaAs; InaTs; dTs; dCs;

m08

UTR

dCs; dAs; dGs; dGs; dAs; dGs;

dGs; InaGs; InaAs; InaA-

Sup

214
FXN-521
GAGGGAAAATGAATT
FXN
Antisense/
human
InaGs; InaAs; InaGs; dGs;

m08

UTR

dGs; dAs; dAs; dAs; dAs; dTs;

dGs; dAs; InaAs; InaTs; InaT-

Sup

215
FXN-522
GAAAATGAATTGTCTTC
FXN
Antisense/
human
InaGs; InaAs; InaAs; dAs;

m08

UTR

dAs; dTs; dGs; dAs; dAs; dTs;

dTs; dGs; dTs; dCs; InaTs;

InaTs; InaC-Sup

216
EPO-37
GGTGGTTTCAGTTCT
EPO
3′
human
dGs; InaGs; dTs; InaGs; dGs;

m02

InaTs; dTs; InaTs; dCs; InaAs;

dGs; InaTs; dTs; InaCs;

dT-Sup

217
EPO-38
TTTTTGGTGGTTTCAGTT
EPO
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m02
CT

InaGs; dGs; InaTs; dGs; InaGs;

dTs; InaTs; dTs; InaCs; dAs;

InaGs; dTs; InaTs; dCs;

InaT-Sup

218
EPO-39
AGCGTGCTATCTGGG
EPO
5′
human
dAs; InaGs; dCs; InaGs; dTs;

m02

InaGs; dCs; InaTs; dAs; InaTs;

dCs; InaTs; dGs; InaGs;

dG-Sup

219
EPO-40
TGGCCCAGGGACTCT
EPO
5′
human
dTs; InaGs; dGs; InaCs; dCs;

m02

InaCs; dAs; InaGs; dGs; InaGs;

dAs; InaCs; dTs; InaCs;

dT-Sup

220
EPO-41
TCTGCGGCTCTGGC
EPO
5′
human
dTs; InaCs; dTs; InaGs; dCs;

m02

InaGs; dGs; InaCs; dTs; InaCs;

dTs; InaGs; dGs; InaC-

Sup

221
EPO-42
CGGTCCGGCTCTGGG
EPO
5′
human
dCs; InaGs; dGs; InaTs; dCs;

m02

InaCs; dGs; InaGs; dCs; InaTs;

dCs; InaTs; dGs; InaGs;

dG-Sup

222
EPO-43
TCATCCCGGGAAGCT
EPO
5′
human
dTs; InaCs; dAs; InaTs; dCs;

m02

InaCs; dCs; InaGs; dGs; InaGs;

dAs; InaAs; dGs; InaCs;

dT-Sup

223
EPO-44
CCCCAAGTCCCCGCT
EPO
5′
human
dCs; InaCs; dCs; InaCs; dAs;

m02

InaAs; dGs; InaTs; dCs; InaCs;

s; dCs; InaCs; dGs; InaCs; dT-

Sup

224
EPO-45
CCAACCATGCAAGCA
EPO
5′
human
dCs; InaCs; dAs; InaAs; dCs;

m02

InaCs; dAs; InaTs; dGs; InaCs;

dAs; InaAs; dGs; InaCs;

dA-Sup

225
EPO-46
TGGCCCAGGGACTCTTC
EPO
5′
human
dTs; InaGs; dGs; InaCs; dCs;

m02

InaCs; dAs; InaGs; dGs; InaGs;

dAs; InaCs; dTs; InaCs;

dTs; InaTs; dC-Sup

226
EPO-47
CGGTCCGGCTCTGGGTT
EPO
5′
human
dCs; InaGs; dGs; InaTs; dCs;

m02
C

InaCs; dGs; InaGs; dCs; InaTs;

dCs; InaTs; dGs; InaGs;

dGs; InaTs; dTs; InaC-Sup

227
EPO-48
CCAACCATGCAAGCACC
EPO
5′
human
dCs; InaCs; dAs; InaAs; dCs;

m02

InaCs; dAs; InaTs; dGs; InaCs;

dAs; InaAs; dGs; InaCs;

dAs; InaCs; dC-Sup

228
EPO-49
TGGCCCAGGGACTCTCA
EPO
5′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
CAAAGTGAC

InaCs; dAs; InaGs; dGs; InaGs;

Gs; dAs; InaCs; dTs; InaCs;

dTs; InaCs; dAs; dCs; dAs; dAs;

dAs; dGs; dTs; InaGs; dAs;

InaC-Sup

229
EPO-50
CGGTCCGGCTCTGGGAA
EPO
5′
human
dCs; InaGs; dGs; InaTs; dCs;

m02
GAAACTTTC

InaCs; dGs; InaGs; dCs; InaTs;

dCs; InaTs; dGs; InaGs; dGs;

InaAs; dAs; dGs; dAs;

dAs; dAs; dCs; dTs; InaTs; dTs;

InaC-Sup

230
EPO-51
CCAACCATGCAAGCACT
EPO
5′
human
dCs; InaCs; dAs; InaAs; dCs;

m02
CAAAGAGTC

InaCs; dAs; InaTs; dGs; InaCs;

dAs; InaAs; dGs; InaCs;

dAs; InaCs; dTs; dCs; dAs; dAs;

dAs; dGs; dAs; InaGs; dTs;

InaC-Sup

231
EPO-52
TGGCCCAGGGACTCTTT
EPO
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
TTGGTGGTTTCAGTTCT

InaCs; dAs; InaGs; dGs; InaGs;

dAs; InaCs; dTs; InaCs;

dTs; InaTs; dTs; InaTs; dTs;

InaGs; dGs; InaTs; dGs; InaGs;

dTs; InaTs; dTs; InaCs; dAs;

InaGs; dTs; InaTs; dCs; InaT-

Sup

232
EPO-53
CGGTCCGGCTCTGGGTT
EPO
5′ and 3′
human
dCs; InaGs; dGs; InaTs; dCs;

m02
TTTGGTGGTTTCAGTTCT

InaCs; dGs; InaGs; dCs; InaTs;

dCs; InaTs; dGs; InaGs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaGs; dTs; InaGs;

dGs; InaTs; dTs; InaTs; dCs;

InaAs; dGs; InaTs; dTs; InaCs;

dT-Sup

233
EPO-54
CCAACCATGCAAGCATT
EPO
5′ and 3′
human
dCs; InaCs; dAs; InaAs; dCs;

m02
TTTGGTGGTTTCAGTTCT

InaCs; dAs; InaTs; dGs; InaCs;

dAs; InaAs; dGs; InaCs; dAs;

InaTs; dTs; InaTs; dTs;

InaTs; dGs; InaGs; dTs; InaGs;

dGs; InaTs; dTs; InaTs; dCs;

InaAs; dGs; InaTs; dTs; InaCs;

dT-Sup

234
EPO-55
CAGGGACTCTTTTTGGT
EPO
5′ and 3′
human
dCs; InaAs; dGs; InaGs; dGs;

m02
GGTTTCA

InaAs; dCs; InaTs; dCs; InaTs;

dTs; InaTs; dTs; InaTs;

dGs; InaGs; dTs; InaGs; dGs;

InaTs; dTs; InaTs; dCs; InaA-

Sup

235
EPO-56
CGGCTCTGGGTTTTTGG
EPO
5′ and 3′
human
dCs; InaGs; dGs; InaCs; dTs;

m02
TGGTTTCA

InaCs; dTs; InaGs; dGs; InaGs;

dTs; InaTs; dTs; InaTs;

dTs; InaGs; dGs; InaTs; dGs;

InaGs; dTs; InaTs; dTs; InaCs;

dA-Sup

236
EPO-57
CATGCAAGCATTTTTGG
EPO
5′ and 3′
human
dCs; InaAs; dTs; InaGs; dCs;

m02
TGGTTTCA

InaAs; dAs; InaGs; dCs; InaAs;

dTs; InaTs; dTs; InaTs;

dTs; InaGs; dGs; InaTs; dGs;

InaGs; dTs; InaTs; dTs; InaCs;

dA-Sup

237
EPO-58
TGGCCCAGGGACTCGGT
EPO
5′ and 3′
human
dTs; InaGs; dGs; InaCs; dCs;

m02
GGTTTCAGTTCT

InaCs; dAs; InaGs; dGs; InaGs;

dAs; InaCs; dTs; InaCs;

dGs; InaGs; dTs; InaGs; dGs;

InaTs; dTs; InaTs; dCs; InaAs;

dGs; InaTs; dTs; InaCs; dT-

Sup

238
EPO-59
CGGTCCGGCTCTGGTGG
EPO
5′ and 3′
human
dCs; InaGs; dGs; InaTs; dCs;

m02
TGGTTTCAGTTCT

InaCs; dGs; InaGs; dCs; InaTs;

dCs; InaTs; dGs; InaGs;

dTs; InaGs; dGs; InaTs; dGs;

InaGs; dTs; InaTs; dTs; InaCs;

dAs; InaGs; dTs; InaTs; dCs;

InaT-Sup

239
EPO-60
CCAACCATGCAAGCAGG
EPO
5′ and 3′
human
dCs; InaCs; dAs; InaAs; dCs;

m02
TGGTTTCAGTTCT

InaCs; dAs; InaTs; dGs; InaCs;

dAs; InaAs; dGs; InaCs;

dAs; InaGs; dGs; InaTs; dGs;

InaGs; dTs; InaTs; dTs; InaCs;

dAs; InaGs; dTs; InaTs; dCs;

InaT-Sup

240
KLF4-31
TTTTTAGATAAAATATTA
KLF4
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m02
TA

InaAs; dGs; InaAs; dTs; InaAs;

dAs; InaAs; dAs; InaTs; dAs;

InaTs; dTs; InaAs; dTs; InaA-

Sup

241
KLF4-32
TTTTTATTCAGATAAAAT
KLF4
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m02
A

InaAs; dTs; InaTs; dCs; InaAs;

dGs; InaAs; dTs; InaAs;

dAs; InaAs; dAs; InaTs; dA-

Sup

242
KLF4-33
TTTTTGGTTTATTTAAAA
KLF4
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m02
CT

InaGs; dGs; InaTs; dTs; InaTs;

dAs; InaTs; dTs; InaTs; dAs;

InaAs; dAs; InaAs; dCs; InaT-

Sup

243
KLF4-34
TTTTTAAATTTATATTAC
KLF4
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m02
AT

InaAs; dAs; InaAs; dTs; InaTs;

dTs; InaAs; dTs; InaAs; dTs;

InaTs; dAs; InaCs; dAs;

InaT-Sup

244
KLF4-35
TTTTTCTTAAATTTATAT
KLF4
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m02
TA

InaCs; dTs; InaTs; dAs; InaAs;

dAs; InaTs; dTs; InaTs; dAs;

InaTs; dAs; InaTs; dTs; InaA-

Sup

245
KLF4-36
TTTTTCACAAAATGTTCA
KLF4
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m02
TT

InaCs; dAs; InaCs; dAs; InaAs;

dAs; InaAs; dTs; InaGs; dTs;

InaTs; dCs; InaAs; dTs; InaT-

Sup

246
KLF4-37
CCTCCGCCTTCTCCC
KLF4
5′
human
dCs; InaCs; dTs; InaCs; dCs;

m02

InaGs; dCs; InaCs; dTs; InaTs;

dCs; InaTs; dCs; InaCs; dC-

Sup

247
KLF4-38
TCTGGTCGGGAAACT
KLF4
5′
human
dTs; InaCs; dTs; InaGs; dGs;

m02

InaTs; dCs; InaGs; dGs; InaGs;

dAs; InaAs; dAs; InaCs;

dT-Sup

248
KLF4-39
GCTACAGCCTTTTCC
KLF4
5′
human
dGs; InaCs; dTs; InaAs; dCs;

m02

InaAs; dGs; InaCs; dCs; InaTs;

dTs; InaTs; dTs; InaCs; dC-

Sup

249
KLF4-40
CCTCCGCCTTCTCCCC
KLF4
5′
human
dCs; InaCs; dTs; InaCs; dCs;

m02

InaGs; dCs; InaCs; dTs; InaTs;

dCs; InaTs; dCs; InaCs; dCs;

InaC-Sup

250
KLF4-41
TCTGGTCGGGAAACTCC
KLF4
5′
human
dTs; InaCs; dTs; InaGs; dGs;

m02

InaTs; dCs; InaGs; dGs; InaGs;

dAs; InaAs; dAs; InaCs;

dTs; InaCs; dC-Sup

251
KLF4-42
GCTACAGCCTTTTCCC
KLF4
5′
human
dGs; InaCs; dTs; InaAs; dCs;

m02

InaAs; dGs; InaCs; dCs; InaTs;

dTs; InaTs; dTs; InaCs; dCs;

InaC-Sup

252
KLF4-43
CCTCCGCCTTCTCCCTCT
KLF4
5′
human
dCs; InaCs; dTs; InaCs; dCs;

m02
TTGATC

InaGs; dCs; InaCs; dTs; InaTs;

dCs; InaTs; dCs; InaCs; dCs;

InaTs; dCs; dTs; dTs; dTs;

dGs; InaAs; dTs; InaC-Sup

253
KLF4-44
TCTGGTCGGGAAACTCA
KLF4
5′
human
dTs; InaCs; dTs; InaGs; dGs;

m02
ATTATTGTC

InaTs; dCs; InaGs; dGs; InaGs;

dAs; InaAs; dAs; InaCs;

dTs; InaCs; dAs; dAs; dTs;

dTs; dAs; dTs; dTs; InaGs; dTs;

InaC-Sup

254
KLF4-45
GCTACAGCCTTTTCCACT
KLF4
5′
human
dGs; InaCs; dTs; InaAs; dCs;

m02
TTGTTC

InaAs; dGs; InaCs; dCs; InaTs;

dTs; InaTs; dTs; InaCs; dCs;

InaAs; dCs; dTs; dTs; dTs;

dGs; InaTs; dTs; InaC-Sup

255
KLF4-46
CCTCCGCCTTCTCCCTTT
KLF4
5′ and 3′
human
dCs; InaCs; dTs; InaCs; dCs;

m02
TTAGATAAAATATTATA

InaGs; dCs; InaCs; dTs; InaTs;

dCs; InaTs; dCs; InaCs; dCs;

InaTs; dTs; InaTs; dTs; InaTs;

dAs; InaGs; dAs; InaTs;

dAs; InaAs; dAs; InaAs; dTs;

InaAs; dTs; InaTs; dAs; InaTs;

dA-Sup

256
KLF4-47
TCTGGTCGGGAAACTTT
KLF4
5′ and 3′
human
dTs; InaCs; dTs; InaGs; dGs;

m02
TTAGATAAAATATTATA

InaTs; dCs; InaGs; dGs; InaGs;

dAs; InaAs; dAs; InaCs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dGs; InaAs; dTs; InaAs;

dAs; InaAs; dAs; InaTs; dAs;

InaTs; dTs; InaAs; dTs; InaA-

Sup

257
KLF4-48
GCTACAGCCTTTTCCTTT
KLF4
5′ and 3′
human
dGs; InaCs; dTs; InaAs; dCs;

m02
TTAGATAAAATATTATA

InaAs; dGs; InaCs; dCs; InaTs;

dTs; InaTs; dTs; InaCs; dCs;

InaTs; dTs; InaTs; dTs; InaTs;

dAs; InaGs; dAs; InaTs;

dAs; InaAs; dAs; InaAs; dTs;

InaAs; dTs; InaTs; dAs; InaTs;

dA-Sup

258
KLF4-49
CCTCCGCCTTCTCCCTTT
KLF4
5′ and 3′
human
dCs; InaCs; dTs; InaCs; dCs;

m02
TTGGTTTATTTAAAACT

InaGs; dCs; InaCs; dTs; InaTs;

dCs; InaTs; dCs; InaCs; dCs;

InaTs; dTs; InaTs; dTs; InaTs;

dGs; InaGs; dTs; InaTs;

dTs; InaAs; dTs; InaTs; dTs;

InaAs; dAs; InaAs; dAs; InaCs;

dT-Sup

259
KLF4-50
TCTGGTCGGGAAACTTT
KLF4
5′ and 3′
human
dTs; InaCs; dTs; InaGs; dGs;

m02
TTGGTTTATTTAAAACT

InaTs; dCs; InaGs; dGs; InaGs;

dAs; InaAs; dAs; InaCs;

dTs; InaTs; dTs; InaTs; dTs;

InaGs; dGs; InaTs; dTs; InaTs;

dAs; InaTs; dTs; InaTs; dAs;

InaAs; dAs; InaAs; dCs; InaT-

Sup

260
KLF4-51
GCTACAGCCTTTTCCTTT
KLF4
5′ and 3′
human
dGs; InaCs; dTs; InaAs; dCs;

m02
TTGGTTTATTTAAAACT

InaAs; dGs; InaCs; dCs; InaTs;

dTs; InaTs; dTs; InaCs; dCs;

InaTs; dTs; InaTs; dTs; InaTs;

dGs; InaGs; dTs; InaTs;

dTs; InaAs; dTs; InaTs; dTs;

InaAs; dAs; InaAs; dAs; InaCs;

dT-Sup

261
KLF4-52
CCTCCGCCTTCTCCCTTT
KLF4
5′ and 3′
human
dCs; InaCs; dTs; InaCs; dCs;

m02
TTAAATTTATATTACAT

InaGs; dCs; InaCs; dTs; InaTs;

dCs; InaTs; dCs; InaCs; dCs;

InaTs; dTs; InaTs; dTs; InaTs;

dAs; InaAs; dAs; InaTs;

dTs; InaTs; dAs; InaTs; dAs;

InaTs; dTs; InaAs; dCs; InaAs;

dT

262
KLF4-53
TCTGGTCGGGAAACTTT
KLF4
5′ and 3′
human
dTs; InaCs; dTs; InaGs; dGs;

m02
TTAAATTTATATTACAT

InaTs; dCs; InaGs; dGs; InaGs;

dAs; InaAs; dAs; InaCs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dAs; InaAs; dTs; InaTs;

dTs; InaAs; dTs; InaAs; dTs;

InaTs; dAs; InaCs; dAs; InaT-

Sup

263
KLF4-54
GCTACAGCCTTTTCCTTT
KLF4
5′ and 3′
human
dGs; InaCs; dTs; InaAs; dCs;

m02
TTAAATTTATATTACAT

InaAs; dGs; InaCs; dCs; InaTs;

dTs; InaTs; dTs; InaCs; dCs;

InaTs; dTs; InaTs; dTs; InaTs;

dAs; InaAs; dAs; InaTs;

dTs; InaTs; dAs; InaTs; dAs;

InaTs; dTs; InaAs; dCs; InaAs;

dT-Sup

264
KLF4-55
GCCTTCTCCCTTTTTAGA
KLF4
5′ and 3′
human
dGs; InaCs; dCs; InaTs; dTs;

m02
TAAAATA

InaCs; dTs; InaCs; dCs; InaCs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dGs; InaAs; dTs; InaAs;

dAs; InaAs; dAs; InaTs;

dA-Sup

dTs; InaCs; dGs; InaGs; dGs;

265
KLF4-56
TCGGGAAACTTTTTAGA
KLF4
5′ and 3′
human
InaAs; dAs; InaAs; dCs; InaTs;

m02
TAAAATA

dTs; InaTs; dTs; InaTs;

dAs; InaGs; dAs; InaTs; dAs;

InaAs; dAs; InaAs; dTs; InaA-

Sup

266
KLF4-57
AGCCTTTTCCTTTTTAGA
KLF4
5′ and 3′
human
dAs; InaGs; dCs; InaCs; dTs;

m02
TAAAATA

InaTs; dTs; InaTs; dCs; InaCs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dGs; InaAs; dTs; InaAs;

dAs; InaAs; dAs; InaTs;

dA-Sup

267
KLF4-58
GCCTTCTCCCTTTTTGGT
KLF4
5′ and 3′
human
dGs; InaCs; dCs; InaTs; dTs;

m02
TTATTTA

InaCs; dTs; InaCs; dCs; InaCs;

dTs; InaTs; dTs; InaTs; dTs;

InaGs; dGs; InaTs; dTs; InaTs;

dAs; InaTs; dTs; InaTs;

dA-Sup

268
KLF4-59
TCGGGAAACTTTTTGGT
KLF4
5′ and 3′
human
dTs; InaCs; dGs; InaGs; dGs;

m02
TTATTTA

InaAs; dAs; InaAs; dCs; InaTs;

dTs; InaTs; dTs; InaTs;

dGs; InaGs; dTs; InaTs; dTs;

InaAs; dTs; InaTs; dTs; InaA-

Sup

269
KLF4-60
AGCCTTTTCCTTTTTGGT
KLF4
5′ and 3′
human
dAs; InaGs; dCs; InaCs; dTs;

m02
TTATTTA

InaTs; dTs; InaTs; dCs; InaCs;

dTs; InaTs; dTs; InaTs; dTs;

InaGs; dGs; InaTs; dTs; InaTs;

dAs; InaTs; dTs; InaTs;

dA-Sup

270
KLF4-61
GCCTTCTCCCTTTTTAAA
KLF4
5′ and 3′
human
dGs; InaCs; dCs; InaTs; dTs;

m02
TTTATAT

InaCs; dTs; InaCs; dCs; InaCs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dAs; InaAs; dTs; InaTs;

dTs; InaAs; dTs; InaAs;

dT-Sup

271
KLF4-62
TCGGGAAACTTTTTAAA
KLF4
5′ and 3′
human
dTs; InaCs; dGs; InaGs; dGs;

m02
TTTATAT

InaAs; dAs; InaAs; dCs; InaTs;

dTs; InaTs; dTs; InaTs;

dAs; InaAs; dAs; InaTs; dTs;

InaTs; dAs; InaTs; dAs; InaT-

Sup

272
KLF4-63
AGCCTTTTCCTTTTTAAA
KLF4
5′ and 3′
human
dAs; InaGs; dCs; InaCs; dTs;

m02
TTTATAT

InaTs; dTs; InaTs; dCs; InaCs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dAs; InaAs; dTs; InaTs;

dTs; InaAs; dTs; InaAs; dT-

Sup

273
ACTB-01
AGGTGTGCACTTTTA
ACTB
3′
human
dAs; InaGs; dGs; InaTs; dGs;

m02

InaTs; dGs; InaCs; dAs; InaCs;

dTs; InaTs; dTs; InaTs;

dA-Sup

274
ACTB-02
TCATTTTTAAGGTGT
ACTB
3′
human
dTs; InaCs; dAs; InaTs; dTs;

m02

InaTs; dTs; InaTs; dAs; InaAs;

dGs; InaGs; dTs; InaGs; dT-

Sup

275
ACTB-03
TTTTTAGGTGTGCACTTT
ACTB
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m02
TA

InaAs; dGs; InaGs; dTs; InaG;

s; dTs; InaGs; dCs; InaAs; dCs;

InaTs; dTs; InaTs; dTs; InaA-

Sup

276
ACTB-04
TTTTTCATTTTTAAGGTG
ACTB
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m02
T

InaCs; dAs; InaTs; dTs; InaTs;

dTs; InaTs; dAs; InaAs; dGs;

InaGs; dTs; InaGs; dT-Sup

277
ACTB-05
CGCGGTCTCGGCGGT
ACTB
5′
human
dCs; InaGs; dCs; InaGs; dGs;

m02

InaTs; dCs; InaTs; dCs; InaGs;

dGs; InaCs; dGs; InaGs;

dT-Sup

278
ACTB-06
ATCATCCATGGTGAG
ACTB
5′
human
dAs; InaTs; dCs; InaAs; dTs;

m02

InaCs; dCs; InaAs; dTs; InaGs;

dGs; InaTs; dGs; InaAs;

dG-Sup

279
ACTB-07
CGCGGTCTCGGCGGTTT
ACTB
5′ and 3′
human
dCs; InaGs; dCs; InaGs; dGs;

m02
TTAGGTGTGCACTTTTA

InaTs; dCs; InaTs; dCs; InaGs;

dGs; InaCs; dGs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dGs; InaGs; dTs; InaGs;

dTs; InaGs; dCs; InaAs; dCs;

InaTs; dTs; InaTs; dTs; InaA-

Sup

280
ACTB-08
ATCATCCATGGTGAGTT
ACTB
5′ and 3′
human
dAs; InaTs; dCs; InaAs; dTs;

m02
TTTAGGTGTGCACTTTTA

InaCs; dCs; InaAs; dTs; InaGs;

dGs; InaTs; dGs; InaAs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dAs; InaGs; dGs; InaTs;

dGs; InaTs; dGs; InaCs; dAs;

InaCs; dTs; InaTs; dTs; InaTs;

dA-Sup

281
ACTB-09
CGCGGTCTCGGCGGTTT
ACTB
5′ and 3′
human
dCs; InaGs; dCs; InaGs; dGs;

m02
TTCATTTTTAAGGTGT

InaTs; dCs; InaTs; dCs; InaGs;

dGs; InaCs; dGs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaCs; dAs; InaTs; dTs; InaTs;

dTs; InaTs; dAs; InaAs; dGs;

InaGs; dTs; InaGs; dT-Sup

282
ACTB-10
ATCATCCATGGTGAGTT
ACTB
5′ and 3′
human
dAs; InaTs; dCs; InaAs; dTs;

m02
TTTCATTTTTAAGGTGT

InaCs; dCs; InaAs; dTs; InaGs;

dGs; InaTs; dGs; InaAs;

dGs; InaTs; dTs; InaTs; dTs;

InaTs; dCs; InaAs; dTs; InaTs;

dTs; InaTs; dTs; InaAs; dAs;

InaGs; dGs; InaTs; dGs; InaT-

Sup

283
ACTB-11
TCTCGGCGGTTTTTAGG
ACTB
5′ and 3′
human
dTs; InaCs; dTs; InaCs; dGs;

m02
TGTGCAC

InaGs; dCs; InaGs; dGs; InaTs;

dTs; InaTs; dTs; InaTs;

dAs; InaGs; dGs; InaTs; dGs;

InaTs; dGs; InaCs; dAs; InaC-

Sup

284
ACTB-12
CCATGGTGAGTTTTTAG
ACTB
5′ and 3′
human
dCs; InaCs; dAs; InaTs; dGs;

m02
GTGTGCAC

InaGs; dTs; InaGs; dAs; InaGs;

dTs; InaTs; dTs; InaTs;

dTs; InaAs; dGs; InaGs; dTs;

InaGs; dTs; InaGs; dCs; InaAs;

dC-Sup

285
ACTB-13
TCTCGGCGGTTTTTCATT
ACTB
5′ and 3′
human
dTs; InaCs; dTs; InaCs; dGs;

m02
TTTAA

InaGs; dCs; InaGs; dGs; InaTs;

dTs; InaTs; dTs; InaTs;

dCs; InaAs; dTs; InaTs; dTs;

InaTs; dTs; InaAs; dA-Sup

286
ACTB-14
CCATGGTGAGTTTTTCA
ACTB
5′ and 3′
human
dCs; InaCs; dAs; InaTs; dGs;

m02
TTTTTAA

InaGs; dTs; InaGs; dAs; InaGs;

dTs; InaTs; dTs; InaTs;

dTs; InaCs; dAs; InaTs; dTs;

InaTs; dTs; InaTs; dAs; InaA-

Sup

287
ACTB-15
CGCGGTCTCGGCGGTA
ACTB
5′ and 3′
human
dCs; InaGs; dCs; InaGs; dGs;

m02
GGTGTGCACTTTTA

InaTs; dCs; InaTs; dCs; InaGs;

dGs; InaCs; dGs; InaGs;

dTs; InaAs; dGs; InaGs; dTs;

InaGs; dTs; InaGs; dCs; InaAs;

dCs; InaTs; dTs; InaTs;

dTs; InaA-Sup

288
ACTB-16
ATCATCCATGGTGAGAG
ACTB
5′ and 3′
human
dAs; InaTs; dCs; InaAs; dTs;

m02
GTGTGCACTTTTA

InaCs; dCs; InaAs; dTs; InaGs;

dGs; InaTs; dGs; InaAs;

dGs; InaAs; dGs; InaGs; dTs;

InaGs; dTs; InaGs; dCs; InaAs;

dCs; InaTs; dTs; InaTs; dTs;

InaA-Sup

289
ACTB-17
CGCGGTCTCGGCGGTTC
ACTB
5′ and 3′
human
dCs; InaGs; dCs; InaGs; dGs;

m02
ATTTTTAAGGTGT

InaTs; dCs; InaTs; dCs; InaGs;

dGs; InaCs; dGs; InaGs;

dTs; InaTs; dCs; InaAs; dTs;

InaTs; dTs; InaTs; dTs; InaAs;

dAs; InaGs; dGs; InaTs; dGs;

InaT-Sup

dAs; InaTs; dCs; InaAs; dTs;

290
ACTB-18
ATCATCCATGGTGAGTC
ACTB
5′ and 3′
human
InaCs; dCs; InaAs; dTs; InaGs;

m02
ATTTTTAAGGTGT

dGs; InaTs; dGs; InaAs;

dGs; InaTs; dCs; InaAs; dTs;

InaTs; dTs; InaTs; dTs; InaAs;

dAs; InaGs; dGs; InaTs; dGs;

InaT-Sup

291
UTRN-
TGGAGCCGAGCGCTG
UTRN
5′
human
dTs; InaGs; dGs; InaAs; dGs;

192 m02

InaCs; dCs; InaGs; dAs; InaGs;

dCs; InaGs; dCs; InaTs;

dG-Sup

292
UTRN-
GGGCCTGCCCCTTTG
UTRN
5′
human
dGs; InaGs; dGs; InaCs; dCs;

193 m02

InaTs; dGs; InaCs; dCs; InaCs;

dCs; InaTs; dTs; InaTs;

dG-Sup

293
UTRN-
CCCCAAGTCACCTGA
UTRN
5′
human
dCs; InaCs; dCs; InaCs; dAs;

194 m02

InaAs; dGs; InaTs; dCs; InaAs;

dCs; InaCs; dTs; InaGs;

dA-Sup

294
UTRN-
GACATCAATACCTAA
UTRN
5′
human
dGs; InaAs; dCs; InaAs; dTs;

195 m02

InaCs; dAs; InaAs; dTs; InaAs;

dCs; InaCs; dTs; InaAs; dA-

Sup

295
UTRN-
AAACTTTACCAAGTC
UTRN
5′
human
dAs; InaAs; dAs; InaCs; dTs;

196 m02

InaTs; dTs; InaAs; dCs; InaCs;

dAs; InaAs; dGs; InaTs; dC-

Sup

296
UTRN-
TGGAGCCGAGCGCTGC
UTRN
5′
human
dTs; InaGs; dGs; InaAs; dGs;

197 m02
C

InaCs; dCs; InaGs; dAs; InaGs;

dCs; InaGs; dCs; InaTs; dGs;

InaCs; dC-Sup

297
UTRN-
GGGCCTGCCCCTTTGCC
UTRN
5′
human
dGs; InaGs; dGs; InaCs; dCs;

198 m02

InaTs; dGs; InaCs; dCs; InaCs;

dCs; InaTs; dTs; InaTs;

dGs; InaCs; dC-Sup

298
UTRN-
CCCCAAGTCACCTGACC
UTRN
5′
human
dCs; InaCs; dCs; InaCs; dAs;

199 m02

InaAs; dGs; InaTs; dCs; InaAs;

dCs; InaCs; dTs; InaGs;

dAs; InaCs; dC-Sup

299
UTRN-
GACATCAATACCTAACC
UTRN
5′
human
dGs; InaAs; dCs; InaAs; dTs;

200 m02

InaCs; dAs; InaAs; dTs; InaAs;

dCs; InaCs; dTs; InaAs; dAs;

InaCs; dC-Sup

300
UTRN-
AAACTTTACCAAGTCCC
UTRN
5′
human
dAs; InaAs; dAs; InaCs; dTs;

201 m02

InaTs; dTs; InaAs; dCs; InaCs;

dAs; InaAs; dGs; InaTs; dCs;

InaCs; dC-Sup

301
UTRN-
TGGAGCCGAGCGCTGG
UTRN
5′
human
dTs; InaGs; dGs; InaAs; dGs;

202
GAAACCAC

InaCs; dCs; InaGs; dAs; InaGs;

m1000

dCs; InaGs; dCs; InaTs;

dGs; InaGs; dGs; dAs; dAs;

dAs; dCs; InaCs; dAs; InaC-

Sup

302
UTRN-
GGGCCTGCCCCTTTGGG
UTRN
5′
human
dGs; InaGs; dGs; InaCs; dCs;

203
AAACCAC

InaTs; dGs; InaCs; dCs; InaCs;

m1000

dCs; InaTs; dTs; InaTs;

dGs; InaGs; dGs; dAs; dAs;

dAs; dCs; InaCs; dAs; InaC-

Sup

303
UTRN-
CCCCAAGTCACCTGAGG
UTRN
5′
human
dCs; InaCs; dCs; InaCs; dAs;

204
AAACCAC

InaAs; dGs; InaTs; dCs; InaAs;

m1000

dCs; InaCs; dTs; InaGs;

dAs; InaGs; dGs; dAs; dAs;

dAs; dCs; InaCs; dAs; InaC-

Sup

304
UTRN-
GACATCAATACCTAAGG
UTRN
5′
human
dGs; InaAs; dCs; InaAs; dTs;

205
AAACCAC

InaCs; dAs; InaAs; dTs; InaAs;

m1000

dCs; InaCs; dTs; InaAs; dAs;

InaGs; dGs; dAs; dAs; dAs;

dCs; InaCs; dAs; InaC-Sup

305
UTRN-
AAACTTTACCAAGTCGG
UTRN
5′
human
dAs; InaAs; dAs; InaCs; dTs;

206
AAACCAC

InaTs; dTs; InaAs; dCs; InaCs;

m1000

dAs; InaAs; dGs; InaTs; dCs;

InaGs; dGs; dAs; dAs; dAs;

dCs; InaCs; dAs; InaC-Sup

306
UTRN-
ACTGCAATATATTTC
UTRN
3′
human
dAs; InaCs; dTs; InaGs; dCs;

207 m02

InaAs; dAs; InaTs; dAs; InaTs;

dAs; InaTs; dTs; InaTs; dC-

Sup

307
UTRN-
GTGTTAAAATTACTT
UTRN
3′
human
dGs; InaTs; dGs; InaTs; dTs;

208 m02

InaAs; dAs; InaAs; dAs; InaTs;

dTs; InaAs; dCs; InaTs;

dT-Sup

308
UTRN-
TTTTTACTGCAATATATT
UTRN
3′
human
dTs; InaTs; dTs; InaTs; dTs;

209 m02
TC

InaAs; dCs; InaTs; dGs; InaCs;

dAs; InaAs; dTs; InaAs; dTs;

InaAs; dTs; InaTs; dTs; InaC-

Sup

309
UTRN-
TTTTTGTGTTAAAATTAC
UTRN
3′
human
dTs; InaTs; dTs; InaTs; dTs;

210 m02
TT

InaGs; dTs; InaGs; dTs; InaTs;

dAs; InaAs; dAs; InaAs; dTs;

InaTs; dAs; InaCs; dTs; InaT-

Sup

310
UTRN-
CCGAGCGCTGTTTTTAC
UTRN
5′ and 3′
human
dCs; InaCs; dGs; InaAs; dGs;

211 m02
TGCAATAT

InaCs; dGs; InaCs; dTs; InaGs;

dTs; InaTs; dTs; InaTs;

dTs; InaAs; dCs; InaTs; dGs;

InaCs; dAs; InaAs; dTs; InaAs;

dT-Sup

311
UTRN-
TGCCCCTTTGTTTTTACT
UTRN
5′ and 3′
human
dTs; InaGs; dCs; InaCs; dCs;

212 m02
GCAATAT

InaCs; dTs; InaTs; dTs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dCs; InaTs; dGs; InaCs;

dAs; InaAs; dTs; InaAs; dT-

Sup

312
UTRN-
AGTCACCTGATTTTTACT
UTRN
5′ and 3′
human
dAs; InaGs; dTs; InaCs; dAs;

213 m02
GCAATAT

InaCs; dCs; InaTs; dGs; InaAs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dCs; InaTs; dGs; InaCs;

dAs; InaAs; dTs; InaAs;

dT-Sup

313
UTRN-
CAATACCTAATTTTTACT
UTRN
5′ and 3′
human
dCs; InaAs; dAs; InaTs; dAs;

214 m02
GCAATAT

InaCs; dCs; InaTs; dAs; InaAs;

dTs; InaTs; dTs; InaTs; dTs;

InaAs; dCs; InaTs; dGs; InaCs;

dAs; InaAs; dTs; InaAs; dT-

Sup

314
UTRN-
TTACCAAGTCTTTTTACT
UTRN
5′ and 3′
human
dTs; InaTs; dAs; InaCs; dCs;

215 m02
GCAATAT

InaAs; dAs; InaGs; dTs; InaCs;

dTs; InaTs; dTs; InaTs;

dTs; InaAs; dCs; InaTs; dGs;

InaCs; dAs; InaAs; dTs; InaAs;

dT-Sup

315
UTRN-
CCGAGCGCTGTTTTTGT
UTRN
5′ and 3′
human
dCs; InaCs; dGs; InaAs; dGs;

216 m02
GTTAAAAT

InaCs; dGs; InaCs; dTs; InaGs;

dTs; InaTs; dTs; InaTs;

dTs; InaGs; dTs; InaGs; dTs;

InaTs; dAs; InaAs; dAs; InaAs;

dT-Sup

316
UTRN-
TGCCCCTTTGTTTTTGTG
UTRN
5′ and 3′
human
dTs; InaGs; dCs; InaCs; dCs;

217 m02
TTAAAAT

InaCs; dTs; InaTs; dTs; InaGs;

dTs; InaTs; dTs; InaTs; dTs;

InaGs; dTs; InaGs; dTs; InaTs;

dAs; InaAs; dAs; InaAs;

dT-Sup

317
UTRN-
AGTCACCTGATTTTTGT
UTRN
5′ and 3′
human
dAs; InaGs; dTs; InaCs; dAs;

218 m02
GTTAAAAT

InaCs; dCs; InaTs; dGs; InaAs;

dTs; InaTs; dTs; InaTs; dTs;

InaGs; dTs; InaGs; dTs; InaTs;

dAs; InaAs; dAs; InaAs; dT-

Sup

318
UTRN-
CAATACCTAATTTTTGTG
UTRN
5′ and 3′
human
dCs; InaAs; dAs; InaTs; dAs;

219 m02
TTAAAAT

InaCs; dCs; InaTs; dAs; InaAs;

dTs; InaTs; dTs; InaTs; dTs;

InaGs; dTs; InaGs; dTs; InaTs;

dAs; InaAs; dAs; InaAs;

dT-Sup

319
UTRN-
TTACCAAGTCTTTTTGTG
UTRN
5′ and 3′
human
dTs; InaTs; dAs; InaCs; dCs;

220 m02
TTAAAAT

InaAs; dAs; InaGs; dTs; InaCs;

dTs; InaTs; dTs; InaTs;

dTs; InaGs; dTs; InaGs; dTs;

InaTs; dAs; InaAs; dAs; InaAs;

dT-Sup

320
HBF-XXX
TGTCTGTAGCTCCAG
HBF
5′
human
dTs; InaGs; dTs; InaCs;

m02

dTs; InaG; dTs; InaA; dGs;

InaC; dTs; InaC; dCs; InaA;

dGs-Sup

321
HBF-XXX
TAGCTCCAGTGAGGC
HBF
5′
human
dTs; InaAs; dGs; InaCs;

m02

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dGs; InaAs; dGs;

InaGs; dC-Sup

322
HBF-XXX
TTTCTTCTCCCACCA
HBF
5′
human
dTs; InaTs; dTs; InaCs; dTs;

m02

InaTs; dCs; InaTs; dCs;

InaCs; dCs; InaAs; dCs;

InaCs; dA-Sup

323
HBF-XXX
TGTCTGTAGCTCCAGC
HBF
5′
human
dTs; InaGs; dTs; InaCs;

m02
C

dTs; InaG; dTs; InaA; dGs;

InaC; dTs; InaC; dCs; InaA;

dGs; InaCs; dC-Sup

324
HBF-XXX
TAGCTCCAGTGAGGC
HBF
5′
human
dTs; InaAs; dGs; InaCs;

m02
CC

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dGs; InaAs; dGs;

InaGs; dC; InaCs; dC-Sup

325
HBF-XXX
TTTCTTCTCCCACCAC
HBF
5′
human
dTs; InaTs; dTs; InaCs; dTs;

m02
C

InaTs; dCs; InaTs; dCs;

InaCs; dCs; InaAs; dCs;

InaCs; dA; InaCs; dC-Sup

326
HBF-XXX
TGTCTGTAGCTCCAG
HBF
5′
human
dTs; InaGs; dTs; InaCs;

m03
GGAAACCAC

dTs; InaG; dTs; InaA; dGs;

InaC; dTs; InaC; dCs; InaA;

dGs; InaGs; dGs; dAs;

dAs; dAs; dCs; InaCs; dAs;

InaC-Sup

327
HBF-XXX
TAGCTCCAGTGAGGC
HBF
5′
human
dTs; InaAs; dGs; InaCs;

m04
GGAAACCAC

dTs; InaCs; dCs; InaAs; dGs;

InaTs; dGs; InaAs; dGs;

InaGs; dC; InaGs; dGs;

dAs; dAs; dAs; dCs; InaCs;

dAs; InaC-Sup

328
HBF-XXX
TTTCTTCTCCCACCAG
HBF
5′
human
dTs; InaTs; dTs; InaCs; dTs;

m05
GAAACCAC

InaTs; dCs; InaTs; dCs;

InaCs; dCs; InaAs; dCs;

InaCs; dA; InaGs; dGs; dAs;

dAs; dAs; dCs; InaCs; dAs;

InaC-Sup

329
HBF-XXX
TTTTTGTGTGATCTCT
HBF
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m06
TAGC

InaGs; dATs; InaGs; dTs;

InaGs; dAs; InaTs; dCs;

InaTs; dCs; InaTs; dTs;

InaAs; dGs; InaC-Sup

330
HBF-XXX
TTTTTGTGATCTCTTA
HBF
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m07
GCAG

InaGs; dTs; InaGs; dAs;

InaTs; dCs; InaTs; dCs;

InaTs; dTs; InaAs; dGs;

InaCs; dAs; InaG-Sup

331
HBF-XXX
TTTTTTGATCTCTTAG
HBF
3′
human
dTs; InaTs; dTs; InaTs; dTs;

m08
CAGA

InaTs; dGs; InaAs; dTs;

InaCs; dTs; InaCs; dTs;

InaTs; dAs; InaGs; dCs;

InaAs; dGs; InaA-Sup

332
SMN-XXX
ATTTCTCTCAATCCT
SMN
5′
human
dAs; InaTs; dTs; InaTs;

m02

dCs; InaT; dCs; InaT; dCs;

InaA; dAs; InaT; dCs; InaC;

dTs-Sup

333
SMN-
GGCGTGTATATTTTT
SMN
5′
human
dGs; InaGs; dCs; InaGs;

XXX

dTs; InaGs; dTs; InaAs; dTs;

m03

InaAs; dTs; InaTs; dTs;

InaTs; dT-Sup

334
SMN-
GGTTATCGCCCTCCC
SMN
5′
human
dGs; InaGs; dTs; InaTs;

XXX

dAs; InaTs; dCs; InaGs; dCs;

m04

InaCs; dCs; InaTs; dCs;

InaCs; dC-Sup

335
SMN-
ACGACTTCCGCCGCC
SMN
5′
human
dAs; InaCs; dGs; InaAs;

XXX

dCs; InaTs; dTs; InaCs; dCs;

m05

InaGs; dCs; InaCs; dGs;

InaCs; dC-Sup

336
SMN-
ATTTCTCTCAATCCTC
SMN
5′
human
dAs; InaTs; dTs; InaTs;

XXX
C

dCs; InaT; dCs; InaT; dCs;

m06

InaA; dAs; InaT; dCs; InaC;

dTs; InaCs; dC-Sup

337
SMN-
GGCGTGTATATTTTTC
SMN
5′
human
dGs; InaGs; dCs; InaGs;

XXX
C

dTs; InaGs; dTs; InaAs; dTs;

m07

InaAs; dTs; InaTs; dTs;

InaTs; dT; InaCs; dC-Sup

338
SMN-
GGTTATCGCCCTCCCC
SMN
5′
human
dGs; InaGs; dTs; InaTs;

XXX
C

dAs; InaTs; dCs; InaGs; dCs;

m08

InaCs; dCs; InaTs; dCs;

InaCs; dC; InaCs; dC-Sup

339
SMN-
ACGACTTCCGCCGCCC
SMN
5′
human
dAs; InaCs; dGs; InaAs;

XXX
C

dCs; InaTs; dTs; InaCs; dCs;

m09

InaGs; dCs; InaCs; dGs;

InaCs; dC; InaCs; dC-Sup

340
SMN-
ATTTCTCTCAATCCTG
SMN
5′
human
dAs; InaTs; dTs; InaTs;

XXX
GAAACCAC

dCs; InaT; dCs; InaT; dCs;

m10

InaA; dAs; InaT; dCs; InaC;

dTs; InaGs; dGs; dAs; dAs;

dAs; dCs; InaCs; dAs;

InaC-Sup

341
SMN-
GGCGTGTATATTTTTG
SMN
5′
human
dGs; InaGs; dCs; InaGs;

XXX
GAAACCAC

dTs; InaGs; dTs; InaAs; dTs;

m11

InaAs; dTs; InaTs; dTs;

InaTs; dT; InaGs; dGs; dAs;

dAs; dAs; dCs; InaCs; dAs;

InaC-Sup

342
SMN-
GGTTATCGCCCTCCCG
SMN
5′
human
dGs; InaGs; dTs; InaTs;

XXX
GAAACCAC

dAs; InaTs; dCs; InaGs; dCs;

m12

InaCs; dCs; InaTs; dCs;

InaCs; dC; InaGs; dGs; dAs;

dAs; dAs; dCs; InaCs;

dAs; InaC-Sup

343
SMN-XXX
ACGACTTCCGCCGCC
SMN
5′
human
dAs; InaCs; dGs; InaAs;

m13
GGAAACCAC

dCs; InaTs; dTs; InaCs; dCs;

InaGs; dCs; InaCs; dGs;

InaCs; dC; InaGs; dGs; dAs;

dAs; dAs; dCs; InaCs;

dAs; InaC-Sup

344
SMN-
TTTTTTAATTTTTTTTT
SMN
3′
human
dTs; InaTs; dTs; InaTs; dTs;

XXX
AAA

InaTs; dAs; InaAs; dTs;

m14

InaTs; dTs; InaTs; dTs; InaTs;

dTs; InaTs; dTs; InaAs;

dAs; InaA-Sup

345
SMN-
TTTTTATATGCAAAAA
SMN
3′
human
dTs; InaTs; dTs; InaTs; dTs;

XXX
AGAA

InaAs; dTs; InaAs; dTs;

m15

InaGs; dCs; InaAs; dAs;

InaAs; dAs; InaAs; dAs;

InaGs; dAs; InaA-Sup

346
SMN-
TTTTTCAAAATATGGG
SMN
3′
human
dTs; InaTs; dTs; InaTs; dTs;

XXX
CCAA

InaCs; dAs; InaAs; dAs;

m16

InaAs; dTs; InaAs; dTs;

InaGs; dGs; InaGs; dCs;

InaCs; dAs; InaA-Sup

Table 7 provides exemplary oligonucleotides for targeting the 5′ and 3′ ends of noncoding RNAs HOTAIR and ANRIL.

TABLE 7

Oligos targeting non-coding RNAs

Target

SEQ
Oligo

Gene
Region (5′

Formatted

ID NO
Name
Base Sequence
Name
or 3′ End)
Organism
Sequence

347
HOTAIR-
TTCACCACATGTAAA
HOTAIR
3′
Human
dTs; InaTs; dCs; InaAs;

1

dCs; InaCs; dAs;

InaCs; dAs; InaTs;

dGs; InaTs; dAs;

InaAs; dA-Sup

348
HOTAIR-
TTTTTTCACCACATGTAA
HOTAIR
3′
Human
dTs; InaTs; dTs; InaTs;

2
A

dTs; InaTs; dCs;

InaAs; dCs; InaCs;

dAs; InaCs; dAs;

InaTs; dGs; InaTs;

dAs; InaAs; dA-

Sup

349
HOTAIR-
AAATCAGGGCAGAATG
HOTAIR
5′
Human
dAs; InaAs; dAs; InaTs;

3
T

dCs; InaAs; dGs;

InaGs; dGs; InaCs;

dAs; InaGs; dAs;

InaAs; dTs; InaGs;

dT-Sup

350
HOTAIR-
AAATCAGGGCAGAATG
HOTAIR
5′
Human
dAs; InaAs; dAs; InaTs;

4
TCC

dCs; InaAs; dGs;

InaGs; dGs; InaCs;

dAs; InaGs; dAs;

InaAs; dTs; InaGs;

dTs; InaCs; dC-

Sup

351
HOTAIR-
AAATCAGGGCAGAATG
HOTAIR
5′
Human
dAs; InaAs; dAs; InaTs;

5
TCCAAAGGTC

dCs; InaAs; dGs;

InaGs; dGs; InaCs;

dAs; InaGs; dAs;

InaAs; dTs; InaGs;

dTs; InaCs; dCs;

InaAs; dAs; InaAs;

dGs; InaGs; dTs;

dC-Sup

352
HOTAIR-
AAATCAGGGCAGAATG
HOTAIR
5′ and 3′
Human
dAs; InaAs; dAs; InaTs;

6
TTTTTTTCACCACATGTA

dCs; InaAs; dGs;

AA

InaGs; dGs; InaCs;

dAs; InaGs; dAs;

InaAs; dTs; InaGs;

dTs; InaTs; dTs;

InaTs; dTs; InaTs;

dTs; InaCs; dAs; InaCs;

dCs; InaAs; dCs;

InaAs; dTs; InaGs;

dTs; InaAs; dAs;

dA-Sup

353
ANRIL-1
TTATTGTCTGAGCCC
ANRIL
3′
Human
dTs; InaTs; dAs; InaTs;

dTs; InaGs; dTs;

InaCs; dTs; InaGs;

dAs; InaGs; dCs;

InaCs; dC-Sup

354
ANRIL-2
TTTTTATTGTCTGAGCCC
ANRIL
3′
Human
dTs; InaTs; dTs; InaTs;

dTs; InaAs; dTs;

InaTs; dGs; InaTs;

dCs; InaTs; dGs;

InaAs; dGs; InaCs;

dCs; dC-Sup

355
ANRIL-3
TCAGGTGACGGATGT
ANRIL
5′
Human
dTs; InaCs; dAs; InaGs;

dGs; InaTs; dGs;

InaAs; dCs; InaGs;

dGs; InaAs; dTs;

InaGs; dT-Sup

356
ANRIL-4
TCAGGTGACGGATGTCC
ANRIL
5′
Human
dTs; InaCs; dAs; InaGs;

dGs; InaTs; dGs;

InaAs; dCs; InaGs;

dGs; InaAs; dTs;

InaGs; dTs; InaCs;

dC-Sup

357
ANRIL-5
TCAGGTGACGGATGTCC
ANRIL
5′
Human
dTs; InaCs; dAs; InaGs;

AAAGGTC

dGs; InaTs; dGs;

InaAs; dCs; InaGs;

dGs; InaAs; dTs;

InaGs; dTs; InaCs;

dCs; InaAs; dAs;

InaAs; dGs; InaGs;

dTs; dC-Sup

358
ANRIL-6
TCAGGTGACGGATGTTT
ANRIL
5′ and 3′
Human
dTs; InaCs; dAs; InaGs;

TTTATTGTCTGAGCCC

dGs; InaTs; dGs;

InaAs; dCs; InaGs;

dGs; InaAs; dTs;

InaGs; dTs; InaTs;

dTs; InaTs; dTs;

InaTs; dAs; InaTs;

dTs; InaGs; dTs;

InaCs; dTs; InaGs;

dAs; InaGs; dCs;

InaCs; dC-Sup

Table 8 provides further exemplary RNA stability oligos for multiple human and mouse genes.

TABLE 8

Oligonucleotides designed to target 5′ and 3′ ends of RNAs

SEQ
Oligo

Target

Formatted

ID NO
Name
Base Sequence
Gene Name
Region
Organism
Sequence

359
FOXP3-
TGTGGGGAGCTCGGC
FOXP3
3′
human
dTs; InaGs; dTs; InaGs;

61 m02

dGs; InaGs; dGs;

InaAs; dGs; InaCs;

dTs; InaCs; dGs; InaGs;

dC-Sup

360
FOXP3-
GGGGAGCTCGGCTGC
FOXP3
3′
human
dGs; InaGs; dGs; InaGs;

62 m02

dAs; InaGs; dCs;

InaTs; dCs; InaGs;

dGs; InaCs; dTs; InaGs;

dC-Sup

361
FOXP3-
TTTTTGTGGGGAGCTC
FOXP3
3′
human
dTs; InaTs; dTs; InaTs;

63 m02
GGC

dTs; InaGs; dTs; InaGs;

dGs; InaGs; dGs;

InaAs; dGs; InaCs;

dTs; InaCs; dGs; InaGs;

dC-Sup

362
FOXP3-
TTTTGGGGAGCTCGGC
FOXP3
3′
human
dTs; InaTs; dTs; InaTs;

64 m02
TGC

dGs; InaGs; dGs; InaGs;

dAs; InaGs; dCs;

InaTs; dCs; InaGs;

dGs; InaCs; dTs;

InaGs; dC-Sup

363
FOXP3-
TTGTCCAAGGGCAGG
FOXP3
5′
human
dTs; InaTs; dGs; InaTs;

65 m02

dCs; InaCs; dAs;

InaAs; dGs; InaGs;

dGs; InaCs; dAs;

InaGs; dG-Sup

364
FOXP3-
TCGATGAGTGTGTGC
FOXP3
5′
human
dTs; InaCs; dGs; InaAs;

66 m02

dTs; InaGs; dAs;

InaGs; dTs; InaGs;

dTs; InaGs; dTs;

InaGs; dC-Sup

365
FOXP3-
AGAAGAAAAACCACG
FOXP3
5′
human
dAs; InaGs; dAs; InaAs;

67 m02

dGs; InaAs; dAs;

InaAs; dAs; InaAs;

dCs; InaCs; dAs; InaCs;

dG-Sup

366
FOXP3-
AATATGATTTCTTCC
FOXP3
5′
human
dAs; InaAs; dTs; InaAs;

68 m02

dTs; InaGs; dAs;

InaTs; dTs; InaTs; dCs;

InaTs; dTs; InaCs;

dC-Sup

367
FOXP3-
GAGATGGGGGACATG
FOXP3
5′
human
dGs; InaAs; dGs; InaAs;

69 m02

dTs; InaGs; dGs;

InaGs; dGs; InaGs;

dAs; InaCs; dAs; InaTs;

dG-Sup

368
PTEN-
TTCAGTTTATTCAAG
PTEN
3′
human
dTs; InaTs; dCs; InaAs;

101 m02

dGs; InaTs; dTs;

InaTs; dAs; InaTs; dTs;

InaCs; dAs; InaAs;

dG-Sup

369
PTEN-
CTGTCTCCACTTTTT
PTEN
3′
human
dCs; InaTs; dGs; InaTs;

102 m02

dCs; InaTs; dCs;

InaCs; dAs; InaCs; dTs;

InaTs; dTs; InaTs;

dT-Sup

370
PTEN-
TGGAATAAAACGGG
PTEN
3′
human
dTs; InaGs; dGs; InaAs;

103 m02

dAs; InaTs; dAs;

InaAs; dAs; InaAs;

dCs; InaGs; dGs; InaG-

Sup

371
PTEN-
ACAATTGAGAAAACA
PTEN
3′
human
dAs; InaCs; dAs; InaAs;

104 m02

dTs; InaTs; dGs;

InaAs; dGs; InaAs;

dAs; InaAs; dAs; InaCs;

dA-Sup

372
PTEN-
CAGTTTTAAGTGGAG
PTEN
3′
human
dCs; InaAs; dGs; InaTs;

105 m02

dTs; InaTs; dTs;

InaAs; dAs; InaGs;

dTs; InaGs; dGs; InaAs;

dG-Sup

373
PTEN-
TGACAAGAATGAGAC
PTEN
3′
human
dTs; InaGs; dAs; InaCs;

106 m02

dAs; InaAs; dGs;

InaAs; dAs; InaTs;

dGs; InaAs; dGs; InaAs;

dC-Sup

374
PTEN-
CCGGGCGAGGGGAGG
PTEN
5′
human
dCs; InaCs; dGs; InaGs;

107 m02

dGs; InaCs; dGs;

InaAs; dGs; InaGs;

dGs; InaGs; dAs; InaGs;

dG-Sup

375
PTEN-
CCGCCGGCCTGCCCG
PTEN
5′
human
dCs; InaCs; dGs; InaCs;

108 m02

dCs; InaGs; dGs;

InaCs; dCs; InaTs;

dGs; InaCs; dCs; InaCs;

dG-Sup

376
PTEN-
CGAGCGCGTATCCTG
PTEN
5′
human
dCs; InaGs; dAs; InaGs;

109 m02

dCs; InaGs; dCs;

InaGs; dTs; InaAs;

dTs; InaCs; dCs; InaTs;

dG-Sup

377
PTEN-
CTGCTTCTCCTCAGC
PTEN
5′
human
dCs; InaTs; dGs; InaCs;

110 m02

dTs; InaTs; dCs;

InaTs; dCs; InaCs; dTs;

InaCs; dAs; InaGs;

dC-Sup

378
PTEN-
TTTTCAGTTTATTCAAG
PTEN
3′
human
dTs; InaTs; dTs; InaTs;

111 m02

dCs; InaAs; dGs; InaTs;

dTs; InaTs; dAs;

InaTs; dTs; InaCs;

dAs; InaAs; dG-Sup

379
PTEN-
TTTTCTGTCTCCACTTTT
PTEN
3′
human
dTs; InaTs; dTs; InaTs;

112 m02
T

dCs; InaTs; dGs;

InaTs; dCs; InaTs; dCs;

InaCs; dAs; InaCs;

dTs; InaTs; dTs; InaTs;

dT-Sup

380
PTEN-
TTTTTGGAATAAAACG
PTEN
3′
human
dTs; InaTs; dTs; InaTs;

113 m02
GG

dTs; InaGs; dGs; InaAs;

dAs; InaTs; dAs;

InaAs; dAs; InaAs;

dCs; InaGs; dGs; InaG-

Sup

381
PTEN-
TTTTACAATTGAGAAAA
PTEN
3′
human
dTs; InaTs; dTs; InaTs;

114 m02
CA

dAs; InaCs; dAs; InaAs;

dTs; InaTs; dGs;

InaAs; dGs; InaAs;

dAs; InaAs; dAs; InaCs;

dA-Sup

382
PTEN-
TTTTCAGTTTTAAGTGG
PTEN
3′
human
dTs; InaTs; dTs; InaTs;

115 m02
AG

dCs; InaAs; dGs; InaTs;

dTs; InaTs; dTs;

InaAs; dAs; InaGs;

dTs; InaGs; dGs; InaAs;

dG-Sup

383
PTEN-
TTTTTGACAAGAATGA
PTEN
3′
human
dTs; InaTs; dTs; InaTs;

116 m02
GAC

dTs; InaGs; dAs; InaCs;

dAs; InaAs; dGs;

InaAs; dAs; InaTs;

dGs; InaAs; dGs;

InaAs; dC-Sup

384
NFE2L2-
AACAGTCATAATAAT
NFE2L2
3′
human
dAs; InaAs; dCs; InaAs;

01 m02

dGs; InaTs; dCs;

InaAs; dTs; InaAs;

dAs; InaTs; dAs;

InaAs; dT-Sup

385
NFE2L2-
TAATTTAACAGTCAT
NFE2L2
3′
human
dTs; InaAs; dAs; InaTs;

02 m02

dTs; InaTs; dAs;

InaAs; dCs; InaAs;

dGs; InaTs; dCs;

InaAs; dT-Sup

386
NFE2L2-
GCACGCTATAAAGCA
NFE2L2
5′
human
dGs; InaCs; dAs; InaCs;

03 m02

dGs; InaCs; dTs;

InaAs; dTs; InaAs;

dAs; InaAs; dGs; InaCs;

dA-Sup

387
NFE2L2-
CCCGGGGCTGGGCTT
NFE2L2
5′
human
dCs; InaCs; dCs; InaGs;

04 m02

dGs; InaGs; dGs;

InaCs; dTs; InaGs;

dGs; InaGs; dCs; InaTs;

dT-Sup

388
NFE2L2-
CCCCGCTCCGCCTCC
NFE2L2
5′
human
dCs; InaCs; dCs; InaCs;

05 m02

dGs; InaCs; dTs;

InaCs; dCs; InaGs;

dCs; InaCs; dTs; InaCs;

dC-Sup

389
NFE2L2-
GCGCCTCCCTGATTT
NFE2L2
5′
human
dGs; InaCs; dGs; InaCs;

06 m02

dCs; InaTs; dCs;

InaCs; dCs; InaTs;

sdG; InaAs; dTs; InaTs;

dT-Sup

390
NFE2L2-
TCGCCGCGGTGGCTG
NFE2L2
5′
human
dTs; InaCs; dGs; InaCs;

07 m02

dCs; InaGs; dCs;

InaGs; dGs; InaTs;

dGs; InaGs; dCs; InaTs;

dG-Sup

391
NFE2L2-
CAGCGAATGGTCGCG
NFE2L2
5′
human
dCs; InaAs; dGs; InaCs;

08 m02

dGs; InaAs; dAs;

InaTs; dGs; InaGs;

dTs; InaCs; dGs; InaCs;

dG-Sup

392
NFE2L2-
TTTTTAACAGTCATAAT
NFE2L2
3′
human
dTs; InaTs; dTs; InaTs;

09 m02
AAT

dTs; InaAs; dAs;

InaCs; dAs; InaGs; dTs;

InaCs; dAs; InaTs;

dAs; InaAs; dTs; InaAs;

dAs; InaT-Sup

393
NFE2L2-
TTTTTAATTTAACAGTC
NFE2L2
3′
human
dTs; InaTs; dTs; InaTs;

10 m02
AT

dTs; InaAs; dAs;

InaTs; dTs; InaTs; dAs;

InaAs; dCs; InaAs;

dGs; InaTs; dCs; InaAs;

dT-Sup

394
ATP2A2-
GCGGCGGCTGCTCTA
ATP2A2
5′
human
dGs; InaCs; dGs; InaGs;

56 m02

dCs; InaGs; dGs;

InaCs; dTs; InaGs;

dCs; InaTs; dCs; InaTs;

dA-Sup

395
ATP2A2-
TTATCGGCCGCTGCC
ATP2A2
5′
human
dTs; InaTs; dAs; InaTs;

34 m02

dCs; InaGs; dGs;

InaCs; dCs; InaGs;

dCs; InaTs; dGs; InaCs;

dC-Sup

396
ATP2A2-
GCGTCGGGGACGGCT
ATP2A2
5′
human
dGs; InaCs; dGs; InaTs;

57 m02

dCs; InaGs; dGs;

InaGs; dGs; InaAs;

dCs; InaGs; dGs; InaCs;

dT-Sup

397
ATP2A2-
GCGGAGGAAACTGCG
ATP2A2
5′
human
dGs; InaCs; dGs; InaGs;

58 m02

dAs; InaGs; dGs;

InaAs; dAs; InaAs;

dCs; InaTs; dGs; InaCs;

dG-Sup

398
ATP2A2-
GCCGCACGCCCGACA
ATP2A2
5′
human
dGs; InaCs; dCs; InaGs;

59 m02

dCs; InaAs; dCs;

InaGs; dCs; InaCs;

dCs; InaGs; dAs; InaCs;

dA-Sup

399
ATP2A2-
CCTGACCCACCCTCC
ATP2A2
5′
human
dCs; InaCs; dTs; InaGs;

60 m02

dAs; InaCs; dCs;

InaCs; dAs; InaCs; dCs;

InaCs; dTs; InaCs;

dC-Sup

400
ATP2A2-
AGGGCAGGCCGCGGC
ATP2A2
5′
human
dAs; InaGs; dGs; InaGs;

61 m02

dCs; InaAs; dGs;

InaGs; dCs; InaCs;

dGs; InaCs; dGs; InaGs;

dC-Sup

401
ATP2A2-
CTGAATCACCCCGCG
ATP2A2
5′
human
dCs; InaTs; dGs; InaAs;

62 m02

dAs; InaTs; dCs;

InaAs; dCs; InaCs; dCs;

InaCs; dGs; InaCs;

dG-Sup

402
ATP2A2-
GGCCCCGAGCTCCGC
ATP2A2
5′
human
dGs; InaGs; dCs; InaCs;

63 m02

dCs; InaCs; dGs;

InaAs; dGs; InaCs;

dTs; InaCs; dCs; InaGs;

dC-Sup

403
ATP2A2-
GCGGCTGCTCTAATA
ATP2A2
5′
human
dGs; InaCs; dGs; InaGs;

64 m02

dCs; InaTs; dGs;

InaCs; dTs; InaCs; dTs;

InaAs; dAs; InaTs;

dA-Sup

404
ATP2A2-
CGCCGCGGCATGTGG
ATP2A2
5′
human
dCs; InaGs; dCs; InaCs;

65 m02

dGs; InaCs; dGs;

InaGs; dCs; InaAs;

dTs; InaGs; dTs; InaGs;

dG-Sup

405
ATP2A2-
CCCTCCTCCTCTTGC
ATP2A2
5′
human
dCs; InaCs; dCs; InaTs;

66 m02

dCs; InaCs; dTs;

InaCs; dCs; InaTs; dCs;

InaTs; dTs; InaGs;

dC-Sup

406
ATP2A2-
GGCCGCGGGCTCGTG
ATP2A2
5′
human
dGs; InaGs; dCs; InaCs;

67 m02

dGs; InaCs; dGs;

InaGs; dGs; InaCs;

dTs; InaCs; dGs; InaTs;

dG-Sup

407
ATP2A2-
GTTATTTTTCTCTGT
ATP2A2
3′
human
dGs; InaTs; dTs; InaAs;

68 m02

dTs; InaTs; dTs;

InaTs; dTs; InaCs; dTs;

InaCs; dTs; InaGs;

dT-Sup

408
ATP2A2-
ATTTAAAATGTTTTA
ATP2A2
3′
human
dAs; InaTs; dTs; InaTs;

69 m02

dAs; InaAs; dAs;

InaAs; dTs; InaGs; dTs;

InaTs; dTs; InaTs;

dA-Sup

409
ATP2A2-
TCTCTGTCCATTTAA
ATP2A2
3′
human
dTs; InaCs; dTs; InaCs;

70 m02

dTs; InaGs; dTs;

InaCs; dCs; InaAs;

dTs; InaTs; dTs; InaAs;

dA-Sup

410
ATP2A2-
TCATTTGGTCATGTG
ATP2A2
3′
human
dTs; InaCs; dAs; InaTs;

71 m02

dTs; InaTs; dGs;

InaGs; dTs; InaCs;

dAs; InaTs; dGs; InaTs;

dG-Sup

411
ATP2A2-
TAGTTCTCTGTACAT
ATP2A2
3′
human
dTs; InaAs; dGs; InaTs;

72 m02

dTs; InaCs; dTs;

InaCs; dTs; InaGs; dTs;

InaAs; dCs; InaAs;

dT-Sup

412
ATP2A2-
TCTGCTGGCTCAACT
ATP2A2
3′
human
dTs; InaCs; dTs; InaGs;

73 m02

dCs; InaTs; dGs;

InaGs; dCs; InaTs; dCs;

InaAs; dAs; InaCs;

dT-Sup

413
ATP2A2-
ATCATAGAATAGATT
ATP2A2
3′
human
dAs; InaTs; dCs; InaAs;

74 m02

dTs; InaAs; dGs;

InaAs; dAs; InaTs;

dAs; InaGs; dAs; InaTs;

dT-Sup

414
ATP2A2-
TTATCATAGAATAGA
ATP2A2
3′
human
dTs; InaTs; dAs; InaTs;

75 m02

dCs; InaAs; dTs;

InaAs; dGs; InaAs;

dAs; InaTs; dAs; InaGs;

dA-Sup

415
ATP2A2-
AATTGACATTTAGCA
ATP2A2
3′
human
dAs; InaAs; dTs; InaTs;

76 m02

dGs; InaAs; dCs;

InaAs; dTs; InaTs; dTs;

InaAs; dGs; InaCs;

dA-Sup

416
ATP2A2-
GACATTTAGCATTTT
ATP2A2
3′
human
dGs; InaAs; dCs; InaAs;

77 m02

dTs; InaTs; dTs;

InaAs; dGs; InaCs;

dAs; InaTs; dTs; InaTs;

dT-Sup

417
ATP2A2-
TTAACCATTCAACAC
ATP2A2
3′
human
dTs; InaTs; dAs; InaAs;

78 m02

dCs; InaCs; dAs;

InaTs; dTs; InaCs; dAs;

InaAs; dCs; InaAs;

dC-Sup

418
mKLF4-
CTTGGCCGGGGAAC
KLF4
5′
mouse
dCs; InaTs; dTs; InaGs;

01 m02
T

dGs; InaCs; dCs;

InaGs; dGs; InaGs;

dGs; InaAs; dAs;

InaCs; dT-Sup

419
mKLF4-
GCCGGGGAACTGCC
KLF4
5′
mouse
dGs; InaCs; dCs; InaGs;

02 m02
G

dGs; InaGs; dGs;

InaAs; dAs; InaCs;

dTs; InaGs; dCs;

InaCs; dG-Sup

420
mKLF4-
CGCCCGGAGCCGCG
KLF4
5′
mouse
dCs; InaGs; dCs; InaCs;

03 m02
C

dCs; InaGs; dGs;

InaAs; dGs; InaCs;

dCs; InaGs; dCs;

InaGs; dC-Sup

421
mKLF4-
CTTGGCCGGGGAAC
KLF4
5′
mouse
dCs; InaTs; dTs; InaGs;

04 m02
TCC

dGs; InaCs; dCs;

InaGs; dGs; InaGs;

dGs; InaAs; dAs;

InaCs; dTs; InaCs;

dC-Sup

422
mKLF4-
GCCGGGGAACTGCC
KLF4
5′
mouse
dGs; InaCs; dCs; InaGs;

05 m02
GC

dGs; InaGs; dGs;

InaAs; dAs; InaCs;

dTs; InaGs; dCs;

InaCs; dGs; InaC-

Sup

423
mKLF4-
CGCCCGGAGCCGCG
KLF4
5′
mouse
dCs; InaGs; dCs; InaCs;

06 m02
CC

dCs; InaGs; dGs;

InaAs; dGs; InaCs;

dCs; InaGs; dCs;

InaGs; dCs; InaC-

Sup

424
mKLF4-
CTTGGCCGGGGAAC
KLF4
5′ and
mouse
dCs; InaTs; dTs; InaGs;

07 m02
TATAAAATTC

3′

dGs; InaCs; dCs;

InaGs; dGs; InaGs;

dGs; InaAs; dAs;

InaCs; dTs; InaAs;

dTs; dAs; dAs; dAs;

dAs; InaTs; dTs;

InaC-Sup

425
mKLF4-
CTTGGCCGGGGAAC
KLF4
5′ and
mouse
dCs; InaTs; dTs; InaGs;

08 m02
TTTTTGTCGTTCAGAT

3′

dGs; InaCs; dCs;

AAAA

InaGs; dGs; InaGs;

dGs; InaAs; dAs;

InaCs; dTs; InaTs;

dTs; InaTs; dTs;

InaGs; dTs; InaCs;

dGs; InaTs; dTs;

InaCs; dAs; InaGs;

dAs; InaTs; dAs; InaAs;

dAs; InaA-Sup

426
mKLF4-
CTTGGCCGGGGAAC
KLF4
5′ and
mouse
dCs; InaTs; dTs; InaGs;

09 m02
TTTTTCAGATAAAAT

3′

dGs; InaCs; dCs;

ATT

InaGs; dGs; InaGs;

dGs; InaAs; dAs;

InaCs; dTs; InaTs;

dTs; InaTs; dTs;

InaCs; dAs; InaGs;

dAs; InaTs; dAs;

InaAs; dAs; InaAs;

dTs; InaAs; dTs;

InaT-Sup

427
mKLF4-
CTTGGCCGGGGAAC
KLF4
5′ and
mouse
dCs; InaTs; dTs; InaGs;

10 m02
TGTCGTTCAGATAAA

3′

dGs; InaCs; dCs;

A

InaGs; dGs; InaGs;

dGs; InaAs; dAs;

InaCs; dTs; InaGs;

dTs; InaCs; dGs;

InaTs; dTs; InaCs;

dAs; InaGs; dAs;

InaTs; dAs; InaAs;

dAs; InaA-Sup

428
mKLF4-
CTTGGCCGGGGAAC
KLF4
5′ and
mouse
dCs; InaTs; dTs; InaGs;

11 m02
TTTCAGATAAAATAT

3′

dGs; InaCs; dCs;

T

InaGs; dGs; InaGs;

dGs; InaAs; dAs;

InaCs; dTs; InaTs;

dTs; InaCs; dAs;

InaGs; dAs; InaTs;

dAs; InaAs; dAs;

InaAs; dTs; InaAs;

dTs; InaT-Sup

429
mKLF4-
CCGGGGAACTTTTTG
KLF4
5′ and
mouse
dCs; InaCs; dGs; InaGs;

12 m02
TCGTTCAGA

3′

dGs; InaGs; dAs;

InaAs; dCs; InaTs;

dTs; InaTs; dTs;

InaTs; dGs; InaTs;

dCs; InaGs; dTs;

InaTs; dCs; InaAs;

dGs; InaA-Sup

430
mKLF4-
CGGGGAACTTTTTCA
KLF4
5′ and
mouse
dCs; InaGs; dGs;

13 m02
GATAAA

3′

InaGs; dGs; InaAs;

dAs; InaCs; dTs;

InaTs; dTs; InaTs;

dTs; InaCs; dAs; InaGs;

dAs; InaTs; dAs;

InaAs; dA-Sup

431
mKLF4-
CGGGGAACTGTCGTT
KLF4
5′ and
mouse
dCs; InaGs; dGs;

14 m02
CAGA

3′

InaGs; dGs; InaAs;

dAs; InaCs; dTs;

InaGs; dTs; InaCs;

dGs; InaTs; dTs; InaCs;

dAs; InaGs; dA-

Sup

432
mKLF4-
CCGGGGAACTTTCAG
KLF4
5′ and
mouse
dCs; InaCs; dGs; InaGs;

15 m02
ATAAA

3′

dGs; InaGs; dAs;

InaAs; dCs; InaTs;

dTs; InaTs; dCs;

InaAs; dGs; InaAs;

dTs; InaAs; dAs;

InaA-Sup

433
mKLF4-
GTCGTTCAGATAAAA
KLF4
3′
mouse
dGs; InaTs; dCs; InaGs;

16 m02

dTs; InaTs; dCs;

InaAs; dGs; InaAs;

dTs; InaAs; dAs;

InaAs; dA-Sup

434
mKLF4-
TTCAGATAAAATATT
KLF4
3′
mouse
dTs; InaTs; dCs; InaAs;

17 m02

dGs; InaAs; dTs;

InaAs; dAs; InaAs;

dAs; InaTs; dAs;

InaTs; dT-Sup

435
mKLF4-
TTTTTGTCGTTCAGAT
KLF4
3′
mouse
dTs; InaTs; dTs; InaTs;

18 m02
AAAA

dTs; InaGs; dTs;

InaCs; dGs; InaTs;

dTs; InaCs; dAs;

InaGs; dAs; InaTs;

dAs; InaAs; dAs;

InaA-Sup

436
mKLF4-
TTTTTCAGATAAAAT
KLF4
3′
mouse
dTs; InaTs; dTs; InaTs;

19 m02
ATT

dTs; InaCs; dAs;

InaGs; dAs; InaTs;

dAs; InaAs; dAs;

InaAs; dTs; InaAs;

dTs; InaT-Sup

437
mFXN-
CTCCGCGGCCGCTCC
FXN
5′
mouse
dCs; InaTs; dCs; InaCs;

01 m02

dGs; InaCs; dGs;

InaGs; dCs; InaCs;

dGs; InaCs; dTs;

InaCs; dC-Sup

438
mFXN-
GCCCACATGCTACTC
FXN
5′
mouse
dGs; InaCs; dCs; InaCs;

02 m02

dAs; InaCs; dAs;

InaTs; dGs; InaCs;

dTs; InaAs; dCs;

InaTs; dC-Sup

439
mFXN-
TCCGAACGCCCACAT
FXN
5′
mouse
dTs; InaCs; dCs; InaGs;

03 m02

dAs; InaAs; dCs;

InaGs; dCs; InaCs;

dCs; InaAs; dCs;

InaAs; dT-Sup

440
mFXN-
CGAGGACTCGGTGG
FXN
5′
mouse
dCs; InaGs; dAs; InaGs;

04 m02
T

dGs; InaAs; dCs;

InaTs; dCs; InaGs;

dGs; InaTs; dGs;

InaGs; dT-Sup

441
mFXN-
CCAGCTCCGCGGCCG
FXN
5′
mouse
dCs; InaCs; dAs; InaGs;

05 m02

dCs; InaTs; dCs;

InaCs; dGs; InaCs;

dGs; InaGs; dCs;

InaCs; dG-Sup

442
mFXN-
CTCCGCGGCCGCTCC
FXN
5′
mouse
dCs; InaTs; dCs; InaCs;

06 m02
C

dGs; InaCs; dGs;

InaGs; dCs; InaCs;

dGs; InaCs; dTs;

InaCs; dCs; InaC-

Sup

443
mFXN-
GCCCACATGCTACTC
FXN
5′
mouse
dGs; InaCs; dCs; InaCs;

07 m02
C

dAs; InaCs; dAs;

InaTs; dGs; InaCs;

dTs; InaAs; dCs;

InaTs; dCs; InaC-

Sup

444
mFXN-
CTCCGCGGCCGCTCC
FXN
5′
mouse
dCs; InaTs; dCs; InaCs;

08 m02
TCAAAGATC

dGs; InaCs; dGs;

InaGs; dCs; InaCs;

dGs; InaCs; dTs;

InaCs; dCs; InaTs;

dCs; dAs; dAs; dAs;

dGs; InaAs; dTs;

InaC-Sup

445
mFXN-
GCCCACATGCTACTC
FXN
5′
mouse
dGs; InaCs; dCs; InaCs;

09 m02
CCAAAGGTC

dAs; InaCs; dAs;

InaTs; dGs; InaCs;

dTs; InaAs; dCs;

InaTs; dCs; InaCs;

dCs; dAs; dAs; dAs;

dGs; InaGs; dTs;

InaC-Sup

446
mFXN-
CTCCGCGGCCGCTCC
FXN
5′ and
mouse
dCs; InaTs; dCs; InaCs;

10 m02
TTTTTGGGAGGGAAC

3′

dGs; InaCs; dGs;

ACACT

InaGs; dCs; InaCs;

dGs; InaCs; dTs;

InaCs; dCs; InaTs;

dTs; InaTs; dTs;

InaTs; dGs; InaGs;

dGs; InaAs; dGs;

InaGs; dGs; InaAs;

dAs; InaCs; dAs;

InaCs; dAs; InaCs;

dT-Sup

447
mFXN-
GCCCACATGCTACTC
FXN
5′ and
mouse
dGs; InaCs; dCs;

11 m02
TTTTTGGGAGGGAAC

3′

InaCs; dAs; InaCs;

ACACT

dAs; InaTs; dGs;

InaCs; dTs; InaAs; dCs;

InaTs; dCs; InaTs;

dTs; InaTs; dTs;

InaTs; dGs; InaGs;

dGs; InaAs; dGs;

InaGs; dGs; InaAs;

dAs; InaCs; dAs;

InaCs; dAs; InaCs; dT-

Sup

448
mFXN-
CTCCGCGGCCGCTCC
FXN
5′ and
mouse
dCs; InaTs; dCs; InaCs;

12 m02
GGGAGGGAACACAC

3′

dGs; InaCs; dGs;

T

InaGs; dCs; InaCs;

dGs; InaCs; dTs;

InaCs; dCs; InaGs;

dGs; InaGs; dAs;

InaGs; dGs; InaGs;

dAs; InaAs; dCs;

InaAs; dCs; InaAs;

dCs; InaT-Sup

449
mFXN-
GCCCACATGCTACTC
FXN
5′ and
mouse
dGs; InaCs; dCs; InaCs;

13 m02
GGGAGGGAACACAC

3′

dAs; InaCs; dAs;

T

InaTs; dGs; InaCs;

dTs; InaAs; dCs;

InaTs; dCs; InaGs;

dGs; InaGs; dAs;

InaGs; dGs; InaGs;

dAs; InaAs; dCs;

InaAs; dCs; InaAs;

dCs; InaT-Sup

450
mFXN-
CGGCCGCTCCGGGA
FXN
5′ and
mouse
dCs; InaGs; dGs;

14 m02
GGGAAC

3′

InaCs; dCs; InaGs;

dCs; InaTs; dCs;

InaCs; dGs; InaGs;

dGs; InaAs; dGs;

InaGs; dGs; InaAs;

dAs; InaC-Sup

451
mFXN-
CATGCTACTCGGGAG
FXN
5′ and
mouse
dCs; InaAs; dTs; InaGs;

15 m02
GGAAC

3′

dCs; InaTs; dAs;

InaCs; dTs; InaCs;

dGs; InaGs; dGs;

InaAs; dGs; InaGs;

dGs; InaAs; dAs;

InaC-Sup

452
mFXN-
GGGAGGGAACACAC
FXN
3′
mouse
dGs; InaGs; dGs;

16 m02
T

InaAs; dGs; InaGs;

dGs; InaAs; dAs;

InaCs; dAs; InaCs;

dAs; InaCs; dT-

Sup

453
mFXN-
GGGGTCTTCACCTGA
FXN
3′
mouse
dGs; InaGs; dGs;

17 m02

InaGs; dTs; InaCs;

dTs; InaTs; dCs;

InaAs; dCs; InaCs;

dTs; InaGs; dA-Sup

454
mFXN-
GGCTGTTATATCATG
FXN
3′
mouse
dGs; InaGs; dCs;

18 m02

InaTs; dGs; InaTs;

dTs; InaAs; dTs;

InaAs; dTs; InaCs;

dAs; InaTs; dG-Sup

455
mFXN-
GGCATTTTAAGATGG
FXN
3′
mouse
dGs; InaGs; dCs;

19 m02

InaAs; dTs; InaTs;

dTs; InaTs; dAs;

InaAs; dGs; InaAs;

dTs; InaGs; dG-Sup

456
mFXN-
TTTTTGGGAGGGAAC
FXN
3′
mouse
dTs; InaTs; dTs;

20 m02
ACACT

InaTs; dTs; InaGs;

dGs; InaGs; dAs;

InaGs; dGs; InaGs;

Ads; InaAs; dCs;

InaAs; dCs; InaAs;

dCs; InaT-Sup

457
mFXN-
TTTTTGGCTGTTATAT
FXN
3′
mouse
dTs; InaTs; dTs;

21 m02
CATG

InaTs; dTs; InaGs;

dGs; InaCs; dTs;

InaGs; dTs; InaTs; dAs;

InaTs; dAs; InaTs;

dCs; InaAs; dTs;

InaG-Sup

Table 9 provides further exemplary RNA 5′ and 3′ end targeting oligos for multiple human and mouse genes.

TABLE 9

Oligonucleotides designed to target 5′ and 3′ ends of RNAs

SEQ
Oligo

Gene
Target

Formatted

ID NO
Name
Base Sequence
Name
Region
Organism
Sequence

459
FXN-654
TGTCTCATTTGGAGA
FXN
3′
human
dTs; InaGs; dTs; InaCs;

m02

dTs; InaCs; dAs; InaTs;

dTs; InaTs; dGs; InaGs;

dAs; InaGs; dA-

Sup

460
FXN-655
ATAATGAAGCTGGG
FXN
3′
human
dAs; InaTs; dAs; InaAs;

m02

dTs; InaGs; dAs; InaAs;

dGs; InaCs; dTs; InaGs;

dGs; InaG-Sup

461
FXN-656
TTTTCCCTCCTGGAA
FXN
3′
human
dTs; InaTs; dTs; InaTs;

m02

dCs; InaCs; dCs; InaTs;

dCs; InaCs; dTs; InaGs;

dGs; InaAs; dA-

Sup

462
FXN-657
TGCATAATGAAGCTG
FXN
3′
human
dTs; InaGs; dCs; InaAs;

m02

dTs; InaAs; dAs; InaTs;

dGs; InaAs; dAs;

InaGs; dCs; InaTs; dG-

Sup

463
FXN-658
AAATCCTTCAAAGAA
FXN
3′
human
dAs; InaAs; dAs; InaTs;

m02

dCs; InaCs; dTs; InaTs;

dCs; InaAs; dAs; InaAs;

dGs; InaAs; dA-

Sup

464
FXN-659
TTGGAAGATTTTTTG
FXN
3′
human
dTs; InaTs; dGs; InaGs;

m02

dAs; InaAs; dGs; InaAs;

dTs; InaTs; dTs;

InaTs; dTs; InaTs; dG-

Sup

465
FXN-660
GCATTCTTGTAGCAG
FXN
3′
human
dGs; InaCs; dAs; InaTs;

m02

dTs; InaCs; dTs; InaTs;

dGs; InaTs; dAs; InaGs;

dCs; InaAs; dG-

Sup

466
FXN-557
ACAACAAAAAACAGA
FXN
3′
human
dAs; InaCs; dAs; InaAs;

m02

dCs; InaAs; dAs; InaAs;

dAs; InaAs; dAs;

InaCs; dAs; InaGs; dA-

Sup

467
FXN-662
TGAAGCTGGGGTCTT
FXN
3′
human
dTs; InaGs; dAs; InaAs;

m02

dGs; InaCs; dTs; InaGs;

dGs; InaGs; dGs;

InaTs; dCs; InaTs; dT-

Sup

468
FXN-663
CCTGAAAACATTTGT
FXN
3′
human
dCs; InaCs; dTs; InaGs;

m02

dAs; InaAs; dAs; InaAs;

dCs; InaAs; dTs; InaTs;

dTs; InaGs; dT-

Sup

469
FXN-664
TTCATTTTCCCTCCT
FXN
3′
human
dTs; InaTs; dCs; InaAs;

m02

dTs; InaTs; dTs; InaTs;

dCs; InaCs; dCs; InaTs;

dCs; InaCs; dT-

Sup

470
FXN-665
TTATTATTATTATAT
FXN
3′
human
dTs; InaTs; dAs; InaTs;

m02

dTs; InaAs; dTs; InaTs;

dAs; InaTs; dTs; InaAs;

dTs; InaAs; dT-

Sup

471
FXN-666
TAACTTTGCATGAAT
FXN
3′
human
dTs; InaAs; dAs; InaCs;

m02

dTs; InaTs; dTs; InaGs;

dCs; InaAs; dTs; InaGs;

dAs; InaAs; dT-

Sup

472
FXN-667
ATACAAACATGTATG
FXN
3′
human
dAs; InaTs; dAs; InaCs;

m02

dAs; InaAs; dAs; InaCs;

dAs; InaTs; dGs; InaTs;

dAs; InaTs; dG-

Sup

473
FXN-668
ATTGTAAACCTATAA
FXN
3′
human
dAs; InaTs; dTs; InaGs;

m02

dTs; InaAs; dAs; InaAs;

dCs; InaCs; dTs; InaAs;

dTs; InaAs; dA-

Sup

474
FXN-669
TGGAGTTGGGGTTAT
FXN
3′
human
dTs; InaGs; dGs; InaAs;

m02

dGs; InaTs; dTs; InaGs;

dGs; InaGs; dGs; InaTs;

dTs; InaAs; dT-

Sup

475
FXN-670
GTTGGGGTTATTTAG
FXN
3′
human
dGs; InaTs; dTs; InaGs;

m02

dGs; InaGs; dGs; InaTs;

dTs; InaAs; dTs;

InaTs; dTs; InaAs; dG-

Sup

476
FXN-671
CTCCGCCCTCCAG
FXN
5′
human
dCs; InaTs; dCs; InaCs;

m02

dGs; InaCs; dCs; InaCs;

dTs; InaCs; dCs; InaAs;

dG-Sup

477
FXN-672
CCGCCCTCCAG
FXN
5′
human
dCs; InaCs; dGs; InaCs;

m02

dCs; InaCs; dTs; InaCs;

dCs; InaAs; dG-Sup

478
FXN-673
GCCCTCCAG
FXN
5′
human
dGs; InaCs; dCs; InaCs;

m02

dTs; InaCs; dCs; InaAs;

dG-Sup

479
FXN-674
CCCGCTCCGCCCTCC
FXN
5′
human
dCs; InaCs; dCs; InaGs;

m02

dCs; InaTs; dCs; InaCs;

dGs; InaCs; dCs; InaCs;

dTs; InaCs; dC-

Sup

480
FXN-675
CGCTCCGCCCTCC
FXN
5′
human
dCs; InaGs; dCs; InaTs;

m02

dCs; InaCs; dGs; InaCs;

dCs; InaCs; dTs; InaCs;

dC-Sup

481
FXN-676
CTCCGCCCTCC
FXN
5′
human
dCs; InaTs; dCs; InaCs;

m02

dGs; InaCs; dCs; InaCs;

dTs; InaCs; dC-Sup

482
FXN-677
CCGCCCTCC
FXN
5′
human
dCs; InaCs; dGs; InaCs;

m02

dCs; InaCs; dTs; InaCs;

dC-Sup

483
FXN-678
GCCACTGGCCGCA
FXN
5′
human
dGs; InaCs; dCs; InaAs;

m02

dCs; InaTs; dGs; InaGs;

dCs; InaCs; dGs;

InaCs; dA-Sup

484
FXN-679
CACTGGCCGCA
FXN
5′
human
dCs; InaAs; dCs; InaTs;

m02

dGs; InaGs; dCs; InaCs;

dGs; InaCs; dA-

Sup

485
FXN-680
GCGACCCCTGGTG
FXN
5′
human
dGs; InaCs; dGs; InaAs;

m02

dCs; InaCs; dCs; InaCs;

dTs; InaGs; dGs;

InaTs; dG-Sup

486
FXN-681
GACCCCTGGTG
FXN
5′
human
dGs; InaAs; dCs; InaCs;

m02

dCs; InaCs; dTs; InaGs;

dGs; InaTs; dG-Sup

487
FXN-682
CTGGCCGCAGGCA
FXN
5′
human
dCs; InaTs; dGs; InaGs;

m02

dCs; InaCs; dGs; InaCs;

dAs; InaGs; dGs;

InaCs; dA-Sup

488
FXN-683
GGCCACTGGCCGC
FXN
5′
human
dGs; InaGs; dCs; InaCs;

m02

dAs; InaCs; dTs; InaGs;

dGs; InaCs; dCs;

InaGs; dC-Sup

489
FXN-684
CTGGTGGCCACTG
FXN
5′
human
dCs; InaTs; dGs; InaGs;

m02

dTs; InaGs; dGs; InaCs;

dCs; InaAs; dCs; InaTs;

dG-Sup

490
FXN-685
GACCCCTGGTGGC
FXN
5′
human
dGs; InaAs; dCs; InaCs;

m02

dCs; InaCs; dTs; InaGs;

dGs; InaTs; dGs;

InaGs; dC-Sup

491
FXN-686
GCGGCGACCCCTG
FXN
5′
human
dGs; InaCs; dGs; InaGs;

m02

dCs; InaGs; dAs;

InaCs; dCs; InaCs; dCs;

InaTs; dG-Sup

492
FXN-687
GTGCTGCGGCGAC
FXN
5′
human
dGs; InaTs; dGs; InaCs;

m02

dTs; InaGs; dCs; InaGs;

dGs; InaCs; dGs;

InaAs; dC-Sup

493
FXN-688
GCTGGGTGCTGCG
FXN
5′
human
dGs; InaCs; dTs; InaGs;

m02

dGs; InaGs; dTs; InaGs;

dCs; InaTs; dGs;

InaCs; dG-Sup

494
FXN-689
CCAGCGCTGGGTG
FXN
5′
human
dCs; InaCs; dAs; InaGs;

m02

dCs; InaGs; dCs; InaTs;

dGs; InaGs; dGs;

InaTs; dG-Sup

495
FXN-690
GCCCTCCAGCGCT
FXN
5′
human
dGs; InaCs; dCs; InaCs;

m02

dTs; InaCs; dCs; InaAs;

dGs; InaCs; dGs;

InaCs; dT-Sup

496
FXN-691
CGCCCGCTCCGCC
FXN
5′
human
dCs; InaGs; dCs; InaCs;

m02

dCs; InaGs; dCs; InaTs;

dCs; InaCs; dGs; InaCs;

dC-Sup

497
FXN-460
CGCCCTCCAGCGCTGTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaCs;

m1000
TTTATTATTTTGCTTTTT

dCs; InaTs; dCs; InaCs;

dAs; InaGs; dCs;

InaGs; dCs; InaTs; dGs;

dT; dT; dT; dT; dT; dAs;

InaTs; dTs; InaAs;

dTs; InaTs; dTs; InaTs;

dGs; InaCs; dTs; InaTs;

dTs; InaTs; dT-Sup

498
FXN-461
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs;

m1000
TTATTATTTTGCTTTTT

dCs; InaCs; dGs; InaCs;

dCs; InaCs; dTs;

InaCs; dCs; InaAs; dGs;

dT; dT; dT; dT; dT; dAs;

InaTs; dTs; InaAs; dTs;

InaTs; dTs; InaTs; dGs;

InaCs; dTs; InaTs;

dTs; InaTs; dT-Sup

499
FXN-523
CAAGTCCAGTTTGGTTT
FXN
3′
human
InaCs; omeAs; InaAs;

m01

omeGs; InaTs; omeCs;

InaCs; omeAs; InaGs;

omeUs; InaTs;

omeUs; InaGs; omeGs;

InaTs; omeUs; InaT-

Sup

500
FXN-524
GAATAGGCCAAGGAAG
FXN
3′
human
InaGs; omeAs; InaAs;

m01
A

omeUs; InaAs; omeGs;

InaGs; omeCs; InaCs;

omeAs; InaAs;

omeGs; InaGs; omeAs;

InaAs; omeGs; InaA-

Sup

501
FXN-525
ATCAAGCATCTTTTCCG
FXN
3′
human
InaAs; omeUs; InaCs;

m01

omeAs; InaAs; omeGs;

InaCs; omeAs; InaTs;

omeCs; InaTs;

omeUs; InaTs; omeUs;

InaCs; omeCs; InaG-

Sup

502
FXN-526
TTAAAACGGGGCTGGG
FXN
3′
human
InaTs; omeUs; InaAs;

m01
C

omeAs; InaAs; omeAs;

InaCs; omeGs;

InaGs; deaGs; InaGs;

omeCs; InaTs; omeGs;

InaGs; omeGs; InaC-

Sup

503
FXN-527
GATAGCTTTTAATGTCC
FXN
3′
human
InaGs; omeAs; InaTs;

m01

omeAs; InaGs; omeCs;

InaTs; omeUs;

InaTs; omeUs; InaAs;

omeAs; InaTs; omeGs;

InaTs; omeCs; InaC-

Sup

504
FXN-528
AGCTGGGGTCTTGGCCT
FXN
3′
human
InaAs; omeGs; InaCs;

m01

omeUs; InaGs; deaGs;

InaGs; omeGs;

InaTs; omeCs; InaTs;

omeUs; InaGs; omeGs;

InaCs; omeCs; InaT-

Sup

505
FXN-529
CCTCAGCTGCATAATGA
FXN
3′
human
InaCs; omeCs; InaTs;

m01

omeCs; InaAs; omeGs;

InaCs; omeUs;

InaGs; omeCs; InaAs;

omeUs; InaAs; omeAs;

InaTs; omeGs; InaA-

Sup

506
FXN-530
CAACAACAAAAAACAGA
FXN
3′
human
InaCs; omeAs; InaAs;

m01

omeCs; InaAs; omeAs;

InaCs; omeAs;

InaAs; omeAs; InaAs;

omeAs; InaAs; omeCs;

InaAs; omeGs; InaA-

Sup

507
FXN-531
AAAAAAATAAACAACAA
FXN
3′
human
InaAs; omeAs; InaAs;

m01

omeAs; InaAs; omeAs;

InaAs; omeUs;

InaAs; omeAs; InaAs;

omeCs; InaAs; omeAs;

InaCs; omeAs; InaA-

Sup

508
FXN-532
CCTCAAAAGCAGGAATA
FXN
3′
human
InaCs; omeCs; InaTs;

m01

omeCs; InaAs; omeAs;

InaAs; omeAs; InaGs;

omeCs; InaAs;

omeGs; InaGs; omeAs;

InaAs; omeUs; InaA-

Sup

509
FXN-533
ACACATAGCCCAACTGT
FXN
3′
human
InaAs; omeCs; InaAs;

m01

omeCs; InaAs; omeUs;

InaAs; omeGs;

InaCs; omeCs; InaCs;

omeAs; InaAs; omeCs;

InaTs; omeGs; InaT-

Sup

510
FXN-534
CTTTCTACAGAGCTGTG
FXN
3′
human
InaCs; omeUs; InaTs;

m01

omeUs; InaCs; omeUs;

InaAs; omeCs;

InaAs; omeGs; InaAs;

omeGs; InaCs; omeUs;

InaGs; omeUs; InaG-

Sup

511
FXN-535
GTAGGAGGCAACACATT
FXN
3′
human
InaGs; omeUs; InaAs;

m01

omeGs; InaGs; omeAs;

InaGs; omeGs;

InaCs; omeAs; InaAs;

omeCs; InaAs; omeCs;

InaAs; omeUs; InaT-

Sup

512
FXN-536
CAGAACTTGGGGGCAA
FXN
3′
human
InaCs; omeAs; InaGs;

m01
G

omeAs; InaAs; omeCs;

InaTs; omeUs;

InaGs; deaGs; InaGs;

deaGs; InaGs; omeCs;

InaAs; omeAs; InaG-

Sup

513
FXN-537
CCATAGAAATTAAAAAT
FXN
3′
human
InaCs; omeCs; InaAs;

m01

omeUs; InaAs; omeGs;

InaAs; omeAs;

InaAs; omeUs; InaTs;

omeAs; InaAs; omeAs;

InaAs; omeAs; InaT-

Sup

514
FXN-538
ACAATCCAAAAAATCTT
FXN
3′
human
InaAs; omeCs; InaAs;

m01

omeAs; InaTs; omeCs;

InaCs; omeAs; InaAs;

omeAs; InaAs;

omeAs; InaAs; omeUs;

InaCs; omeUs; InaT-

Sup

515
FXN-539
GTGAGGGAGGAAATCC
FXN
3′
human
InaGs; omeUs; InaGs;

m01
G

omeAs; InaGs; omeGs;

InaGs; omeAs;

InaGs; omeGs; InaAs;

omeAs; InaAs; omeUs;

InaCs; omeCs; InaG-

Sup

516
FXN-540
AAGATAAGGGGTATCAT
FXN
3′
human
InaAs; omeAs; InaGs;

m01

omeAs; InaTs; omeAs;

InaAs; omeGs;

InaGs; omeGs; InaGs;

omeUs; InaAs; omeUs;

InaCs; omeAs; InaT-

Sup

517
FXN-541
GGCATAAGACATTATAA
FXN
3′
human
InaGs; omeGs; InaCs;

m01

omeAs; InaTs; omeAs;

InaAs; omeGs; InaAs;

omeCs; InaAs;

omeUs; InaTs; omeAs;

InaTs; omeAs; InaA-

Sup

518
FXN-542
TGTTATATTCAGGTATA
FXN
3′
human
InaTs; omeGs; InaTs;

m01

omeUs; InaAs; omeUs;

InaAs; omeUs;

InaTs; omeCs; InaAs;

omeGs; InaGs; omeUs;

InaAs; omeUs; InaA-

Sup

519
FXN-543
TTTGCTTTTTTAAAGGT
FXN
3′
human
InaTs; omeUs; InaTs;

m01

omeGs; InaCs; omeUs;

InaTs; omeUs; InaTs;

omeUs; InaTs;

omeAs; InaAs; omeAs;

InaGs; omeGs; InaT-

Sup

520
FXN-544
TTTTTCCTTCTTATTAT
FXN
3′
human
InaTs; omeUs; InaTs;

m01

omeUs; InaTs; omeCs;

InaCs; omeUs; InaTs;

omeCs; InaTs; omeUs;

InaAs; omeUs; InaTs;

omeAs; InaT-

Sup

521
FXN-545
CATTTTCCCTCCTGGAA
FXN
3′
human
InaCs; omeAs; InaTs;

m01

omeUs; InaTs; omeUs;

InaCs; omeCs; InaCs;

omeUs; InaCs; omeCs;

InaTs; omeGs;

InaGs; omeAs; InaA-

Sup

522
FXN-546
GAAGAGTGAAGACAAT
FXN
3′
human
InaGs; omeAs; InaAs;

m01
T

omeGs; InaAs; omeGs;

InaTs; omeGs; InaAs;

omeAs; InaGs; omeAs;

InaCs; omeAs;

InaAs; omeUs; InaT-

Sup

523
FXN-547
TAAATCCTTCAAAGAAT
FXN
3′
human
InaTs; omeAs; InaAs;

m01

omeAs; InaTs; omeCs;

InaCs; omeUs; InaTs;

omeCs; InaAs;

omeAs; InaAs; omeGs;

InaAs; omeAs; InaT-

Sup

524
FXN-548
TCATGTACTTCTTGCAG
FXN
3′
human
InaTs; omeCs; InaAs;

m01

omeUs; InaGs; omeUs;

InaAs; omeCs; InaTs;

omeUs; InaCs; omeUs;

InaTs; omeGs;

InaCs; omeAs; InaG-

Sup

525
FXN-549
GGTTGACCAGCTGCTCT
FXN
3′
human
InaGs; omeGs; InaTs;

m01

omeUs; InaGs; omeAs;

InaCs; omeCs; InaAs;

omeGs; InaCs; omeUs;

InaGs; omeCs;

InaTs; omeCs; InaT-

Sup

526
FXN-550
AGATAGAACAGTGAGC
FXN
3′
human
InaAs; omeGs; InaAs;

m01
A

omeUs; InaAs; omeGs;

InaAs; omeAs; InaCs;

omeAs; InaGs;

omeUs; InaGs; omeAs;

InaGs; omeCs; InaA-

Sup

527
FXN-551
TAATGTGTCTCATTTGG
FXN
3′
human
InaTs; omeAs; InaAs;

m01

omeUs; InaGs; omeUs;

InaGs; omeUs; InaCs;

omeUs; InaCs;

omeAs; InaTs; omeUs;

InaTs; omeGs; InaG-

Sup

528
FXN-552
ATTTGTAGGCTACCCTT
FXN
3′
human
InaAs; omeUs; InaTs;

m01

omeUs; InaGs; omeUs;

InaAs; omeGs; InaGs;

omeCs; InaTs;

omeAs; InaCs; omeCs;

InaCs; omeUs; InaT-

Sup

529
FXN-553
GAAAGAAGCCTGAAAA
FXN
3′
human
InaGs; omeAs; InaAs;

m01
C

omeAs; InaGs; omeAs;

InaAs; omeGs; InaCs;

omeCs; InaTs;

omeGs; InaAs; omeAs;

InaAs; omeAs; InaC-

Sup

530
FXN-554
AGAAGTGCTTACACTTT
FXN
3′
human
InaAs; omeGs; InaAs;

m01

omeAs; InaGs; omeUs;

InaGs; omeCs; InaTs;

omeUs; InaAs;

omeCs; InaAs; omeCs;

InaTs; omeUs; InaT-

Sup

531
FXN-555
TCAATGCTAAAGAGCTC
FXN
3′
human
InaTs; omeCs; InaAs;

m01

omeAs; InaTs; omeGs;

InaCs; omeUs; InaAs;

omeAs; InaAs;

omeGs; InaAs; omeGs;

InaCs; omeUs; InaC-

Sup

532
Apoa1_
AGTCTGGGTGTCC
Apoa1
5′
mouse
InaAs; dGs; InaTs;

mus-01

dCs; InaTs; dGs; InaGs;

m12

dGs; InaTs; dGs;

InaTs; dCs; InaC-

Sup

533
Apoa1_
CCGACAGTCTGGG
Apoa1
5′
mouse
InaCs; dCs; InaGs;

mus-02

dAs; InaCs; dAs; InaGs;

m12

dTs; InaCs; dTs;

InaGs; dGs; InaG-

Sup

534
Apoa1_
CTCCGACAGTCTG
Apoa1
5′
mouse
InaCs; dTs; InaCs;

mus-03

dCs; InaGs; dAs; InaCs;

m12

dAs; InaGs; dTs;

InaCs; dTs; InaG-

Sup

535
Apoa1_
GACAGTCTGGGTG
Apoa1
5′
mouse
InaGs; dAs; InaCs;

mus-04

dAs; InaGs; dTs; InaCs;

m12

dTs; InaGs; dGs;

InaGs; dTs; InaG-

Sup

536
Apoa1_
CAGTCTGGGTG
Apoa1
5′
mouse
InaCs; dAs; InaGs;

mus-05

dTs; InaCs; dTs; InaGs;

dGs; InaGs; dTs;

m12

InaG-Sup

537
Apoa1_
CTCAGCCTGGCCCTG
Apoa1
5′
mouse
InaCs; dTs; InaCs;

mus-06

dAs; InaGs; dCs; InaCs;

m12

dTs; InaGs; dGs;

InaCs; dCs; InaCs;

dTs; InaG-Sup

538
Apoa1_
AGTTCAAGGATCAGC
Apoa1
5′
mouse
InaAs; dGs; InaTs;

mus-07

dTs; InaCs; dAs; InaAs;

m12

dGs; InaGs; dAs;

InaTs; dCs; InaAs;

dGs; InaC-Sup

539
Apoa1_
GCTCTCCGACAGTCT
Apoa1
5′
mouse
InaGs; dCs; InaTs;

mus-08

dCs; InaTs; dCs; InaCs;

m12

dGs; InaAs; dCs;

InaAs; dGs; InaTs;

dCs; InaT-Sup

540
Apoa1_
TCTCCGACAGTCT
Apoa1
5′
mouse
InaTs; dCs; InaTs;

mus-09

dCs; InaCs; dGs; InaAs;

m12

dCs; InaAs; dGs;

InaTs; dCs; InaT-

Sup

541
Apoa1_
TCCGACAGTCT
Apoa1
5′
mouse
InaTs; dCs; InaCs;

mus-10

dGs; InaAs; dCs; InaAs;

m12

dGs; InaTs; dCs;

InaT-Sup

542
Apoa1_
CGGAGCTCTCCGACA
Apoa1
5′
mouse
InaCs; dGs; InaGs;

mus-11

dAs; InaGs; dCs;

m12

InaTs; dCs; InaTs; dCs;

InaCs; dGs; InaAs;

dCs; InaA-Sup

543
Apoa1_
GAGCTCTCCGACA
Apoa1
5′
mouse
InaGs; dAs; InaGs;

mus-12

dCs; InaTs; dCs; InaTs;

m12

dCs; InaCs; dGs;

InaAs; dCs; InaA-

Sup

544
Apoa1_
GCTCTCCGACA
Apoa1
5′
mouse
InaGs; dCs; InaTs;

mus-13

dCs; InaTs; dCs; InaCs;

m12

dGs; InaAs; dCs;

InaA-Sup

545
Apoa1_
CTATTCCATTTTGGA
Apoa1
3′
mouse
InaCs; dTs; InaAs;

mus-14

dTs; InaTs; dCs; InaCs;

m12

dAs; InaTs; dTs;

InaTs; dTs; InaGs;

dGs; InaA-Sup

546
Apoa1_
CTATTCCATTTTG
Apoa1
3′
mouse
InaCs; dTs; InaAs;

mus-15

dTs; InaTs; dCs; InaCs;

m12

dAs; InaTs; dTs;

InaTs; dTs; InaG-

Sup

547
Apoa1_
ATTCCATTTTGGAAA
Apoa1
3′
mouse
InaAs; dTs; InaTs;

mus-16

dCs; InaCs; dAs; InaTs;

m12

dTs; InaTs; dTs;

InaGs; dGs; InaAs;

dAs; InaA-Sup

548
Apoa1_
CCATTTTGGAAAGGT
Apoa1
3′
mouse
InaCs; dCs; InaAs;

mus-17

dTs; InaTs; dTs; InaTs;

m12

dGs; InaGs; dAs;

InaAs; dAs; InaGs;

dGs; InaT-Sup

549
Apoa1_
CCATTTTGGAAAG
Apoa1
3′
mouse
InaCs; dCs; InaAs;

mus-18

dTs; InaTs; dTs; InaTs;

m12

dGs; InaGs; dAs;

InaAs; dAs; InaG-

Sup

550
Apoa1_
CATTTTGGAAAGGTT
Apoa1
3′
mouse
InaCs; dAs; InaTs;

mus-19

dTs; InaTs; dTs; InaGs;

m12

dGs; InaAs; dAs;

InaAs; dGs; InaGs;

dTs; InaT-Sup

551
Apoa1_
CATTTTGGAAAGG
Apoa1
3′
mouse
InaCs; dAs; InaTs;

mus-20

dTs; InaTs; dTs; InaGs;

m12

dGs; InaAs; dAs;

InaAs; dGs; InaG-

Sup

552
Apoa1_
GGAAAGGTTTATTGT
Apoa1
3′
mouse
InaGs; dGs; InaAs;

mus-21

dAs; InaAs; dGs;

m12

InaGs; dTs; InaTs;

dTs; InaAs; dTs; InaTs;

dGs; InaT-Sup

InaTs; dCs; dCs; InaGs;

553
Apoa1_
TCCGACAGTCTCCATT
Apoa1
5′ and 3′
mouse
dAs; dCs; InaAs;

mus-22
TTGGAA

dGs; dTs; InaCs; dTs;

m22

dCs; InaCs; dAs;

dTs; InaTs; dTs; dTs;

InaGs; dGs; dAs; InaA-

Sup

554
Apoa1_
GCTCTCCGACACCATT
Apoa1
5′ and 3′
mouse
InaGs; dCs; dTs; InaCs;

mus-23
TTGGAA

dTs; dCs; InaCs;

m22

dGs; dAs; InaCs;

dAs; dCs; InaCs; dAs;

dTs; InaTs; dTs; dTs;

InaGs; dGs; dAs; InaA-

Sup

555
Apoa1_
TCCGACAGTCTCATTT
Apoa1
5′ and 3′
mouse
InaTs; dCs; dCs; InaGs;

mus-24
TGGAAA

dAs; dCs; InaAs;

m22

dGs; dTs; InaCs;

dTs; dCs; InaAs; dTs;

dTs; InaTs; dTs; dGs;

InaGs; dAs; dAs; InaA-

Sup

556
Apoa1_
GCTCTCCGACACATTT
Apoa1
5′ and 3′
mouse
InaGs; dCs; dTs; InaCs;

mus-25
TGGAAA

dTs; dCs; InaCs;

m22

dGs; dAs; InaCs; dAs;

dCs; InaAs; dTs;

dTs; InaTs; dTs; dGs;

InaGs; dAs; dAs;

InaA-Sup

557
FXN-761
CCTCAAAAGCAGGAA
FXN
3′
human
InaCs; omeCs; InaTs;

m01

omeCs; InaAs;

omeAs; InaAs; omeAs;

InaGs; omeCs;

InaAs; omeGs; InaGs;

omeAs; InaA-

Sup

558
FXN-762
CCTCAAAAGCAGG
FXN
3′
human
InaCs; omeCs; InaTs;

m01

omeCs; InaAs;

omeAs; InaAs; omeAs;

InaGs; omeCs;

InaAs; omeGs; InaG-

Sup

559
FXN-763
CCTCAAAAGCA
FXN
3′
human
InaCs; omeCs; InaTs;

m01

omeCs; InaAs;

omeAs; InaAs; omeAs;

InaGs; omeCs; InaA-

Sup

560
FXN-764
TCAAAAGCAGGAA
FXN
3′
human
InaTs; omeCs; InaAs;

m01

omeAs; InaAs;

omeAs; InaGs; omeCs;

InaAs; omeGs;

InaGs; omeAs; InaA-

Sup

561
FXN-765
CAAAAGCAGGA
FXN
3′
human
InaCs; omeAs; InaAs;

m01

omeAs; InaAs;

omeGs; InaCs; omeAs;

InaGs; omeGs;

InaA-Sup

562
FXN-766
CCGCCCTCCAGCCTCA
FXN
5′ and 3′
human
InaCs; omeCs; InaGs;

m01
AAAGCAGGAAT

omeCs; InaCs;

omeCs; InaTs;

omeCs; InaCs; omeAs;

InaGs; omeCs;

InaCs; omeTs; InaCs;

omeAs; InaAs; omeAs;

InaAs; omeGs;

InaCs; omeAs; InaGs;

omeGs; InaAs;

omeAs; InaT-Sup

563
FXN-767
CCGCCCTCCAGCCTCA
FXN
5′ and 3′
human
InaCs; omeCs; InaGs;

m01
AAAGCAGGA

omeCs; InaCs;

omeCs; InaTs; omeCs;

InaCs; omeAs;

InaGs; omeCs; InaCs;

omeTs; InaCs;

omeAs; InaAs; omeAs;

InaAs; omeGs;

InaCs; omeAs; InaGs;

omeGs; InaA-

Sup

564
FXN-768
CCGCCCTCCAGCCTCA
FXN
5′ and 3′
human
InaCs; omeCs; InaGs;

m01
AAAGCAG

omeCs; InaCs;

omeCs; InaTs; omeCs;

InaCs; omeAs;

InaGs; omeCs; InaCs;

omeTs; InaCs;

omeAs; InaAs; omeAs;

InaAs; omeGs;

InaCs; omeAs; InaG-

Sup

565
FXN-769
CCGCCCTCCAGCCTCA
FXN
5′ and 3′
human
InaCs; omeCs; InaGs;

m01
AAAGC

omeCs; InaCs;

omeCs; InaTs; omeCs;

InaCs; omeAs;

InaGs; omeCs; InaCs;

omeTs; InaCs;

omeAs; InaAs; omeAs;

InaAs; omeGs;

InaC-Sup

566
FXN-770
GCCCTCCAGCCTCAAA
FXN
5′ and 3′
human
InaGs; omeCs; InaCs;

m01
AGCAGGAAT

omeCs; InaTs;

omeCs; InaCs; omeAs;

InaGs; omeCs;

InaCs; omeTs; InaCs;

omeAs; InaAs;

omeAs; InaAs; omeGs;

InaCs; omeAs;

InaGs; omeGs; InaAs;

omeAs; InaT-

Sup

567
FXN-771
GCCCTCCAGCCTCAAA
FXN
5′ and 3′
human
InaGs; omeCs; InaCs;

m01
AGCAGGA

omeCs; InaTs;

omeCs; InaCs; omeAs;

InaGs; omeCs;

InaCs; omeTs; InaCs;

omeAs; InaAs;

omeAs; InaAs; omeGs;

InaCs; omeAs;

InaGs; omeGs; InaA-

Sup

568
FXN-772
GCCCTCCAGCCTCAAA
FXN
5′ and 3′
human
InaGs; omeCs; InaCs;

m01
AGCAG

omeCs; InaTs;

omeCs; InaCs; omeAs;

InaGs; omeCs;

InaCs; omeTs; InaCs;

omeAs; InaAs;

omeAs; InaAs; omeGs;

InaCs; omeAs;

InaG-Sup

569
FXN-773
GCCCTCCAGCCTCAAA
FXN
5′ and 3′
human
InaGs; omeCs; InaCs;

m01
AGC

omeCs; InaTs;

omeCs; InaCs; omeAs;

InaGs; omeCs;

InaCs; omeTs; InaCs;

omeAs; InaAs;

omeAs; InaAs; omeGs;

InaC-Sup

570
FXN-774
CCCTCCAGCCTCAAAA
FXN
5′ and 3′
human
InaCs; omeCs; InaCs;

m01
G

omeTs; InaCs;

omeCs; InaAs; omeGs;

InaCs; omeCs;

InaTs; omeCs; InaAs;

omeAs; InaAs;

omeAs; InaG-Sup

571
FXN-776
CCTCCAGCCTCAAAA
FXN
5′ and 3′
human
InaCs; omeCs; InaTs;

m01

omeCs; InaCs;

omeAs; InaGs; omeCs;

InaCs; omeTs;

InaCs; omeAs; InaAs;

omeAs; InaA-

Sup

572
FXN-777
GCCCTCCAGTCAAAA
FXN
5′ and 3′
human
InaGs; omeCs; InaCs;

m01
GCAGGA

omeCs; InaTs;

omeCs; InaCs; omeAs;

InaGs; omeTs;

InaCs; omeAs; InaAs;

omeAs; InaAs;

omeGs; InaCs; omeAs;

InaGs; omeGs;

InaA-Sup

573
FXN-778
GCCCTCCAGCAAAAG
FXN
5′ and 3′
human
InaGs; omeCs; InaCs;

m01
CAGG

omeCs; InaTs;

omeCs; InaCs; omeAs;

InaGs; omeCs;

InaAs; omeAs; InaAs;

omeAs; InaGs;

omeCs; InaAs; omeGs;

InaG-Sup

574
FXN-779
CCGCCCTCCAGTCAAA
FXN
5′ and 3′
human
InaCs; omeCs; InaGs;

m01
AGCAGGA

omeCs; InaCs;

omeCs; InaTs; omeCs;

InaCs; omeAs;

InaGs; omeTs; InaCs;

omeAs; InaAs;

omeAs; InaAs; omeGs;

InaCs; omeAs;

InaGs; omeGs; InaA-

Sup

575
FXN-780
CCGCCCTCCAGCAAA
FXN
5′ and 3′
human
InaCs; omeCs; InaGs;

m01
AGCAGG

omeCs; InaCs;

omeCs; InaTs; omeCs;

InaCs; omeAs;

InaGs; omeCs; InaAs;

omeAs; InaAs;

omeAs; InaGs; omeCs;

InaAs; omeGs;

InaG-Sup

576
FXN-671
CTCCGCCCTCCAG
FXN
5′
human
InaCs; omeTs; InaCs;

m01

omeCs; InaGs;

omeCs; InaCs; omeCs;

InaTs; omeCs;

InaCs; omeAs; InaG-

Sup

577
FXN-672
CCGCCCTCCAG
FXN
5′
human
InaCs; omeCs; InaGs;

m01

omeCs; InaCs;

omeCs; InaTs;

omeCs; InaCs; omeAs;

InaG-Sup

578
FXN-673
GCCCTCCAG
FXN
5′
human
InaGs; omeCs; InaCs;

m01

omeCs; InaTs;

omeCs; InaCs; omeAs;

InaG-Sup

579
FXN-674
CCCGCTCCGCCCTCC
FXN
5′
human
InaCs; omeCs; InaCs;

m01

omeGs; InaCs;

omeTs; InaCs; omeCs;

InaGs; omeCs;

InaCs; omeCs; InaTs;

omeCs; InaC-

Sup

580
FXN-675
CGCTCCGCCCTCC
FXN
5′
human
InaCs; omeGs; InaCs;

m01

omeTs; InaCs;

omeCs; InaGs; omeCs;

InaCs; omeCs;

InaTs; omeCs; InaC-

Sup

581
FXN-676
CTCCGCCCTCC
FXN
5′
human
InaCs; omeTs; InaCs;

m01

omeCs; InaGs;

omeCs; InaCs;

omeCs; InaTs; omeCs;

InaC-Sup

582
FXN-677
CCGCCCTCC
FXN
5′
human
InaCs; omeCs; InaGs;

m01

omeCs; InaCs;

omeCs; InaTs;

omeCs; InaC-Sup

583
CD247-
GCCTTTGAGAAAGCA
CD247
5′
human
dGs; InaCs; dCs; InaTs;

90 m02

dTs; InaTs; dGs;

InaAs; dGs; InaAs;

dAs; InaAs; dGs;

InaCs; dA-Sup

584
CD247-
GACTGTGGGGCCTTT
CD247
5′
human
dGs; InaAs; dCs;

91 m02

InaTs; dGs; InaTs;

dGs; InaGs; dGs; InaGs;

dCs; InaCs; dTs;

InaTs; dT-Sup

585
CD247-
AGGAAGTGGAGGACT
CD247
5′
human
dAs; InaGs; dGs; InaAs;

92 m02

dAs; InaGs; dTs;

InaGs; dGs; InaAs;

dGs; InaGs; dAs;

InaCs; dT-Sup

586
CD247-
TGCATTTTCACTGAA
CD247
3′
human
dTs; InaGs; dCs; InaAs;

93 m02

dTs; InaTs; dTs;

InaTs; dCs; InaAs;

dCs; InaTs; dGs; InaAs;

dA-Sup

587
CD247-
CATTTTCACTGAAGC
CD247
3′
human
dCs; InaAs; dTs; InaTs;

94 m02

dTs; InaTs; dCs;

InaAs; dCs; InaTs;

dGs; InaAs; dAs;

InaGs; dC-Sup

588
CD247-
ACTGAAGCATTTATT
CD247
3′
human
dAs; InaCs; dTs; InaGs;

95 m02

dAs; InaAs; dGs;

InaCs; dAs; InaTs;

dTs; InaTs; dAs;

InaTs; dT-Sup

589
CFTR-84
CACACAAATGTATGG
CFTR
3′
human
dCs; InaAs; dCs; InaAs;

m02

dCs; InaAs; dAs;

InaAs; dTs; InaGs;

dTs; InaAs; dTs;

InaGs; dG-Sup

590
CFTR-85
GGATTTTATTGACAA
CFTR
3′
human
dGs; InaGs; dAs; InaTs;

m02

dTs; InaTs; dTs;

InaAs; dTs; InaTs;

dGs; InaAs; dCs;

InaAs; dA-Sup

591
CFTR-86
AAAACAACAAAGTTT
CFTR
3′
human
dAs; InaAs; dAs; InaAs;

m02

dCs; InaAs; dAs;

InaCs; dAs; InaAs;

dAs; InaGs; dTs;

InaTs; dT-Sup

592
CFTR-87
AGTGCCATAAAAAGT
CFTR
3′
human
dAs; InaGs; dTs; InaGs;

m02

dCs; InaCs; dAs;

InaTs; dAs; InaAs;

dAs; InaAs; dAs;

InaGs; dT-Sup

593
CFTR-88
TCAAATATAAAAATT
CFTR
3′
human
dTs; InaCs; dAs; InaAs;

m02

dAs; InaTs; dAs;

InaTs; dAs; InaAs;

dAs; InaAs; dAs;

InaTs; dT-Sup

594
CFTR-89
TTCCCCCCACCCACC
CFTR
3′
human
dTs; InaTs; dCs; InaCs;

m02

dCs; InaCs; dCs;

InaCs; dAs; InaCs;

dCs; InaCs; dAs;

InaCs; dC-Sup

595
CFTR-90
CATTTGCTTCCAATT
CFTR
5′
human
dCs; InaAs; dTs; InaTs;

m02

dTs; InaGs; dCs;

InaTs; dTs; InaCs;

dCs; InaAs; dAs;

InaTs; dT-Sup

596
CFTR-91
GCTCAACCCTTTTTC
CFTR
5′
human
dGs; InaCs; dTs; InaCs;

m02

dAs; InaAs; dCs;

InaCs; dCs; InaTs;

dTs; InaTs; dTs;

InaTs; dC-Sup

597
CFTR-92
AGACCTACTACTCTG
CFTR
5′
human
dAs; InaGs; dAs; InaCs;

m02

dCs; InaTs; dAs;

InaCs; dTs; InaAs;

dCs; InaTs; dCs;

InaTs; dG-Sup

598
FMR1-
CCCTCCACCGGAAGT
FMR1
5′
human
dCs; InaCs; dCs; InaTs;

58 m02

dCs; InaCs; dAs;

InaCs; dCs; InaGs;

dGs; InaAs; dAs;

InaGs; dT-Sup

599
FMR1-
GCCCGCGCTCGCCGT
FMR1
5′
human
dGs; InaCs; dCs; InaCs;

59 m02

dGs; InaCs; dGs;

InaCs; dTs; InaCs;

dGs; InaCs; dCs;

InaGs; dT-Sup

600
FMR1-
ACGCCCCCTGGCAGC
FMR1
5′
human
dAs; InaCs; dGs; InaCs;

60 m02

dCs; InaCs; dCs;

InaCs; dTs; InaGs;

dGs; InaCs; dAs;

InaGs; dC-Sup

601
FMR1-
GCTCAGCCCCTCGGC
FMR1
5′
human
dGs; InaCs; dTs; InaCs;

61 m02

dAs; InaGs; dCs;

InaCs; dCs; InaCs; dTs;

InaCs; dGs; InaGs;

dC-Sup

602
FMR1-
AGCAGAGGAAGATCA
FMR1
3′
human
dAs; InaGs; dCs; InaAs;

62 m02

dGs; InaAs; dGs;

InaGs; dAs; InaAs;

dGs; InaAs; dTs;

InaCs; dA-Sup

603
FMR1-
CAGAGGAAGATCAAA
FMR1
3′
human
dCs; InaAs; dGs; InaAs;

63 m02

dGs; InaGs; dAs;

InaAs; dGs; InaAs;

dTs; InaCs; dAs;

InaAs; dA-Sup

604
FMR1-
CAGATTTTTGAAACT
FMR1
3′
human
dCs; InaAs; dGs; InaAs;

64 m02

dTs; InaTs; dTs;

InaTs; dTs; InaGs;

dAs; InaAs; dAs;

InaCs; dT-Sup

605
FMR1-
CAGACTAATTTTTTG
FMR1
3′
human
dCs; InaAs; dGs; InaAs;

65 m02

dCs; InaTs; dAs;

InaAs; dTs; InaTs;

dTs; InaTs; dTs;

InaTs; dG-Sup

606
FMR1-
TTTTTGCTTTTTCAT
FMR1
3′
human
dTs; InaTs; dTs; InaTs;

66 m02

dTs; InaGs; dCs;

InaTs; dTs; InaTs;

dTs; InaTs; dCs; InaAs;

dT-Sup

607
FMR1-
AATTTTTTGCTTTTT
FMR1
3′
human
dAs; InaAs; dTs; InaTs;

67 m02

dTs; InaTs; dTs;

InaTs; dGs; InaCs;

dTs; InaTs; dTs; InaTs;

dT-Sup

608
FMR1-
ATGTTTGGCAATACT
FMR1
3′
human
dAs; InaTs; dGs; InaTs;

68 m02

dTs; InaTs; dGs;

InaGs; dCs; InaAs;

dAs; InaTs; dAs;

InaCs; dT-Sup

609
FMR1-
TTGGCAATACTTTTT
FMR1
3′
human
dTs; InaTs; dGs; InaGs;

69 m02

dCs; InaAs; dAs;

InaTs; dAs; InaCs;

dTs; InaTs; dTs; InaTs;

dT-Sup

610
LAMA1-
GCTGCCCTGGCCCCG
LAMA1
5′
human
dGs; InaCs; dTs; InaGs;

105

dCs; InaCs; dCs;

m02

InaTs; dGs; InaGs;

dCs; InaCs; dCs; InaCs;

dG-Sup

611
LAMA1-
CGGACACACCCCTCG
LAMA1
5′
human
dCs; InaGs; dGs; InaAs;

106

dCs; InaAs; dCs;

m02

InaAs; dCs; InaCs;

dCs; InaCs; dTs;

InaCs; dG-Sup

612
LAMA1-
ACGGGACGCGAGTCC
LAMA1
5′
human
dAs; InaCs; dGs; InaGs;

107

dGs; InaAs; dCs;

m02

InaGs; dCs; InaGs;

dAs; InaGs; dTs;

InaCs; dC-Sup

613
LAMA1-
GTCTGGGGAGAAAGC
LAMA1
5′
human
dGs; InaTs; dCs; InaTs;

108

dGs; InaGs; dGs;

m02

InaGs; dAs; InaGs;

dAs; InaAs; dAs;

InaGs; dC-Sup

614
LAMA1-
CCACTCGGTGGGTCT
LAMA1
5′
human
dCs; InaCs; dAs; InaCs;

109

dTs; InaCs; dGs;

m02

InaGs; dTs; InaGs;

dGs; InaGs; dTs;

InaCs; dT-Sup

615
LAMA1-
TGATCTGTTATCATC
LAMA1
5′
human
dTs; InaGs; dAs; InaTs;

110

dCs; InaTs; dGs;

m02

InaTs; dTs; InaAs;

dTs; InaCs; dAs; InaTs;

dC-Sup

616
LAMA1-
CTGTTATCATCTGTA
LAMA1
3′
human
dCs; InaTs; dGs; InaTs;

111

dTs; InaAs; dTs;

m02

nIaCs; dAs; InaTs;

dCs; InaTs; dGs; InaTs;

dA-Sup

617
LAMA1-
GTGTATAAAGATTTT
LAMA1
3′
human
dGs; InaTs; dGs; InaTs;

112

dAs; InaTs; dAs;

m02

InaAs; dAs; InaGs;

dAs; InaTs; dTs;

InaTs; dT-Sup

618
LAMA1-
CAATTTACATTTTAG
LAMA1
3′
human
dCs; InaAs; dAs; InaTs;

113

dTs; InaTs; dAs;

m02

InaCs; dAs; InaTs;

dTs; InaTs; dTs; InaAs;

dG-Sup

619
LAMA1-
TACATTTTAGACCAT
LAMA1
3′
human
dTs; InaAs; dCs; InaAs;

114

dTs; InaTs; dTs;

m02

InaTs; dAs; InaGs;

dAs; InaCs; dCs; InaAs;

dT-Sup

620
MBNL1-
TGCTATAAGATGTAA
MBNL1
5′
human
dTs; InaGs; dCs; InaTs;

73 m02

dAs; InaTs; dAs;

InaAs; dGs; InaAs;

dTs; InaGs; dTs;

InaAs; dA-Sup

621
MBNL1-
AAGGAAGCCGGCAA
MBNL1
5′
human
dAs; InaAs; dGs; InaGs;

74 m02
G

dAs; InaAs; dGs;

InaCs; dCs; InaGs;

dGs; InaCs; dAs;

InaAs; dG-Sup

622
MBNL1-
CGCCACAACTCATTC
MBNL1
5′
human
dCs; InaGs; dCs; InaCs;

75 m02

dAs; InaCs; dAs;

InaAs; dCs; InaTs;

dCs; InaAs; dTs; InaTs;

dC-Sup

623
MBNL1-
ATGGGAGCATTGTGG
MBNL1
5′
human
dAs; InaTs; dGs; InaGs;

76 m02

dGs; InaAs; dGs;

InaCs; dAs; InaTs;

dTs; InaGs; dTs; InaGs;

dG-Sup

624
MBNL1-
CGCCCGCCCAGCCCC
MBNL1
5′
human
dCs; InaGs; dCs; InaCs;

77 m02

dCs; InaGs; dCs;

InaCs; dCs; InaAs;

dGs; InaCs; dCs; InaCs;

dC-Sup

625
MBNL1-
CCCCTCCCCCGCCCG
MBNL1
5′
human
dCs; InaCs; dCs; InaCs;

78 m02

dTs; InaCs; dCs;

InaCs; dCs; InaCs;

dGs; InaCs; dCs; InaCs;

dG-Sup

626
MBNL1-
CTTCCGCTGCTGCTG
MBNL1
5′
human
dCs; InaTs; dTs; InaCs;

79 m02

dCs; InaGs; dCs;

InaTs; dGs; InaCs;

dTs; InaGs; dCs; InaTs;

dG-Sup

627
MBNL1-
CTTCTTAGTACCAAC
MBNL1
5′
human
dCs; InaTs; dTs; InaCs;

80 m02

dTs; InaTs; dAs;

InaGs; dTs; InaAs;

dCs; InaCs; dAs; InaAs;

dC-Sup

628
MBNL1-
TTTAGAGCAAAATCG
MBNL1
5′
human
dTs; InaTs; dTs; InaAs;

81 m02

dGs; InaAs; dGs;

InaCs; dAs; InaAs;

dAs; InaAs; dTs; InaCs;

dG-Sup

629
MBNL1-
GGTAGTTAAATGTTT
MBNL1
5′
human
dGs; InaGs; dTs; InaAs;

82 m02

dGs; InaTs; dTs;

InaAs; dAs; InaAs;

dTs; InaGs; dTs; InaTs;

dT-Sup

630
MBNL1-
TACTTAAGAAAGAGA
MBNL1
3′
human
dTs; InaAs; dCs; InaTs;

83 m02

dTs; InaAs; dAs;

InaGs; dAs; InaAs;

dAs; InaGs; dAs; InaGs;

dA-Sup

631
MBNL1-
TATACTTAAGAAAGA
MBNL1
3′
human
dTs; InaAs; dTs; InaAs;

84 m02

dCs; InaTs; dTs;

InaAs; dAs; InaGs;

dAs; InaAs; dAs; InaGs;

dA-Sup

632
MECP2-
CGCCGCCGACGCCGG
MECP2
5′
human
dCs; InaGs; dCs; InaCs;

61 m02

dGs; InaCs; dCs;

InaGs; dAs; InaCs;

dGs; InaCs; dCs; InaGs;

dG-Sup

633
MECP2-
CTCTCTCCGAGAGGA
MECP2
5′
human
dCs; InaTs; dCs; InaTs;

62 m02

dCs; InaTs; dCs;

InaCs; dGs; InaAs;

dGs; InaAs; dGs; InaGs;

dA-Sup

634
MECP2-
CGCCCCGCCCTCTTG
MECP2
5′
human
dCs; InaGs; dCs; InaCs;

63 m02

dCs; InaCs; dGs;

InaCs; dCs; InaCs;

dTs; InaCs; dTs; InaTs;

dG-Sup

635
MECP2-
CCGCGCGCTGCTGCA
MECP2
5′
human
dCs; InaCs; dGs; InaCs;

64 m02

dGs; InaCs; dGs;

InaCs; dTs; InaGs;

dCs; InaTs; dGs; InaCs;

dA-Sup

636
MECP2-
CACTTTCACAGAGAG
MECP2
3′
human
dCs; InaAs; dCs; InaTs;

65 m02

dTs; InaTs; dCs;

InaAs; dCs; InaAs;

dGs; InaAs; dGs; InaAs;

dG-Sup

637
MECP2-
CTTTCACATGTATTAA
MECP2
3′
human
dCs; InaTs; dTs; InaTs;

66 m02

dCs; InaAs; dCs;

InaAs; dTs; InaGs;

dTs; InaAs; dTs; InaTs;

dAs; dA-Sup

638
MECP2-
ATGTATTAAAAAACT
MECP2
3′
human
dAs; InaTs; dGs; InaTs;

67 m02

dAs; InaTs; dTs;

InaAs; dAs; InaAs;

dAs; InaAs; dAs; InaCs;

dT-Sup

639
MECP2-
GACATTTTTATGTAA
MECP2
3′
human
dGs; InaAs; dCs; InaAs;

68 m02

dTs; InaTs; dTs;

InaTs; dTs; InaAs;

dTs; InaGs; dTs;

InaAs; dA-Sup

640
MECP2-
CATTTTTATGTAAAT
MECP2
3′
human
dCs; InaAs; dTs; InaTs;

69 m02

dTs; InaTs; dTs;

InaAs; dTs; InaGs;

dTs; InaAs; dAs; InaAs;

dT-Sup

641
MECP2-
AAATTTATAAGGCAA
MECP2
3′
human
dAs; InaAs; dAs; InaTs;

70 m02

dTs; InaTs; dAs;

InaTs; dAs; InaAs;

dGs; InaGs; dCs;

InaAs; dA-Sup

642
MECP2-
AGGCAAACTCTTTAT
MECP2
3′
human
dAs; InaGs; dGs; InaCs;

71 m02

dAs; InaAs; dAs;

InaCs; dTs; InaCs;

dTs; InaTs; dTs;

InaAs; dT-Sup

643
MECP2-
GTCTCTGGAACAATT
MECP2
3′
human
dGs; InaTs; dCs; InaTs;

72 m02

dCs; InaTs; dGs;

InaGs; dAs; InaAs;

dCs; InaAs; dAs;

InaTs; dT-Sup

644
MECP2-
CAGTTCAAACACAGA
MECP2
3′
human
dCs; InaAs; dGs; InaTs;

73 m02

dTs; InaCs; dAs;

InaAs; dAs; InaCs;

dAs; InaCs; dAs; InaGs;

dA-Sup

645
MECP2-
CAAACACAGAAGAGA
MECP2
3′
human
dCs; InaAs; dAs; InaAs;

74 m02

dCs; InaAs; dCs;

InaAs; dGs; InaAs;

dAs; InaGs; dAs;

InaGs; dA-Sup

646
MECP2-
AACACAGAAGAGATT
MECP2
3′
human
dAs; InaAs; dCs; InaAs;

75 m02

dCs; InaAs; dGs;

InaAs; dAs; InaGs;

dAs; InaGs; dAs;

InaTs; dT-Sup

647
MECP2-
GGGGGAGAAGAAAG
MECP2
3′
human
dGs; InaGs; dGs; InaGs;

76 m02
G

dGs; InaAs; dGs;

InaAs; dAs; InaGs;

dAs; InaAs; dAs;

InaGs; dG-Sup

648
MECP2-
TCGTTTTTTTTTCTT
MECP2
3′
human
dTs; InaCs; dGs; InaTs;

77 m02

dTs; InaTs; dTs;

InaTs; dTs; InaTs;

dTs; InaTs; dCs; InaTs;

dT-Sup

649
MECP2-
CTTTTTTTTCTTTTT
MECP2
3′
human
dCs; InaTs; dTs; InaTs;

78 m02

dTs; InaTs; dTs;

InaTs; dTs; InaCs;

dTs; InaTs; dTs; InaTs;

dT-Sup

650
MECP2-
CCTATGCTATGGTTA
MECP2
3′
human
dCs; InaCs; dTs; InaAs;

79 m02

dTs; InaGs; dCs;

InaTs; dAs; InaTs;

dGs; InaGs; dTs;

InaTs; dA-Sup

651
MECP2-
AGTTTACTGAAAGAA
MECP2
3′
human
dAs; InaGs; dTs; InaTs;

80 m02

dTs; InaAs; dCs;

InaTs; dGs; InaAs;

dAs; InaAs; dGs;

InaAs; dA-Sup

652
MECP2-
ACTGAAAGAAAAAAA
MECP2
3′
human
dAs; InaCs; dTs; InaGs;

81 m02

dAs; InaAs; dAs;

InaGs; dAs; InaAs;

dAs; InaAs; dAs;

InaAs; dA-Sup

653
MERTK-
CCTTATTCATATTTT
MERTK
3′
human
dCs; InaCs; dTs; InaTs;

66 m02

dAs; InaTs; dTs;

InaCs; dAs; InaTs;

dAs; InaTs; dTs;

InaTs; dT-Sup

654
MERTK-
CTTCCTTATTCATAT
MERTK
3′
human
dCs; InaTs; dTs; InaCs;

67 m02

dCs; InaTs; dTs;

InaAs; dTs; InaTs;

dCs; InaAs; dTs; InaAs;

dT-Sup

655
MERTK-
CAATCCTTCAATATT
MERTK
3′
human
dCs; InaAs; dAs; InaTs;

68 m02

dCs; InaCs; dTs;

InaTs; dCs; InaAs;

dAs; InaTs; dAs;

InaTs; dT-Sup

656
MERTK-
GGCATTTCATTTTAC
MERTK
3′
human
dGs; InaGs; dCs; InaAs;

69 m02

dTs; InaTs; dTs;

InaCs; dAs; InaTs;

dTs; InaTs; dTs;

InaAs; dC-Sup

657
MERTK-
CATTTTACAAATATT
MERTK
3′
human
dCs; InaAs; dTs; InaTs;

70 m02

dTs; InaTs; dAs;

InaCs; dAs; InaAs;

dAs; InaTs; dAs;

InaTs; dT-Sup

658
MERTK-
GAAATGAAATAAGTA
MERTK
3′
human
dGs; InaAs; dAs; InaAs;

71 m02

dTs; InaGs; dAs;

InaAs; dAs; InaTs;

dAs; InaAs; dGs;

InaTs; dA-Sup

659
MERTK-
AGATATGCAAGATAA
MERTK
3′
human
dAs; InaGs; dAs; InaTs;

72 m02

dAs; InaTs; dGs;

InaCs; dAs; InaAs;

dGs; InaAs; dTs;

InaAs; dA-Sup

660
MERTK-
GCGGGCCCAGCAGGT
MERTK
5′
human
dGs; InaCs; dGs; InaGs;

73 m02

dGs; InaCs; dCs;

InaCs; dAs; InaGs;

dCs; InaAs; dGs;

InaGs; dT-Sup

661
MERTK-
CAGTGAGTGCCGAGT
MERTK
5′
human
dCs; InaAs; dGs; InaTs;

74 m02

dGs; InaAs; dGs;

InaTs; dGs; InaCs;

dCs; InaGs; dAs;

InaGs; dT-Sup

662
MERTK-
GCCCGGGCAGTGAGT
MERTK
5′
human
dGs; InaCs; dCs; InaCs;

75 m02

dGs; InaGs; dGs;

InaCs; dAs; InaGs;

dTs; InaGs; dAs;

InaGs; dT-Sup

663
MERTK-
TGTCCGGGCGGCCCG
MERTK
5′
human
dTs; InaGs; dTs; InaCs;

76 m02

dCs; InaGs; dGs;

InaGs; dCs; InaGs;

dGs; InaCs; dCs;

InaCs; dG-Sup

664
SSPN-47
CGCGCGTGTGCGAGT
SSPN
5′
human
dCs; InaGs; dCs; InaGs;

m02

dCs; InaGs; dTs;

InaGs; dTs; InaGs;

dCs; InaGs; dAs;

InaGs; dT-Sup

665
SSPN-48
CTTCAGACAGGCTGC
SSPN
5′
human
dCs; InaTs; dTs; InaCs;

m02

dAs; InaGs; dAs;

InaCs; dAs; InaGs;

dGs; InaCs; dTs;

InaGs; dC-Sup

666
SSPN-49
ACCTCTGCACTTCAG
SSPN
5′
human
dAs; InaCs; dCs; InaTs;

m02

dCs; InaTs; dGs;

InaCs; dAs; InaCs;

dTs; InaTs; dCs;

InaAs; dG-Sup

667
SSPN-50
CGGCGCGGGTCCCTT
SSPN
5′
human
dCs; InaGs; dGs; InaCs;

m02

dGs; InaCs; dGs;

InaGs; dGs; InaTs;

dCs; InaCs; dCs;

InaTs; dT-Sup

668
SSPN-51
TGGTATTCGAATTAT
SSPN
5′
human
dTs; InaGs; dGs; InaTs;

m02

dAs; InaTs; dTs;

InaCs; dGs; InaAs;

dAs; InaTs; dTs;

InaAs; dT-Sup

669
SSPN-52
CGGCCTGCCCTGGTA
SSPN
5′
human
dCs; InaGs; dGs; InaCs;

m02

dCs; InaTs; dGs;

InaCs; dCs; InaCs;

dTs; InaGs; dGs;

InaTs; dA-Sup

670
SSPN-53
TCAGAGATTATGAAA
SSPN
3′
human
dTs; InaCs; dAs; InaGs;

m02

dAs; InaGs; dAs;

InaTs; dTs; InaAs;

dTs; InaGs; dAs;

InaAs; dA-Sup

671
SSPN-54
TGTTTTCAGAGATTA
SSPN
3′
human
dTs; InaGs; dTs; InaTs;

m02

dTs; InaTs; dCs;

InaAs; dGs; InaAs;

dGs; InaAs; dTs;

InaTs; dA-Sup

672
SSPN-55
CATGTAGAAATGCTT
SSPN
3′
human
dCs; InaAs; dTs; InaGs;

m02

dTs; InaAs; dGs;

InaAs; dAs; InaAs;

dTs; InaGs; dCs;

InaTs; dT-Sup

673
SSPN-56
AAACATGTAGAAATG
SSPN
3′
human
dAs; InaAs; dAs; InaCs;

m02

dAs; InaTs; dGs;

InaTs; dAs; InaGs;

dAs; InaAs; dAs;

InaTs; dG-Sup

674
SSPN-57
TTGATACCATTTATG
SSPN
3′
human
dTs; InaTs; dGs; InaAs;

m02

dTs; InaAs; dCs;

InaCs; dAs; InaTs;

dTs; InaTs; dAs; InaTs;

dG-Sup

675
SSPN-58
GAACTCAATTATTAT
SSPN
3′
human
dGs; InaAs; dAs; InaCs;

m02

dTs; InaCs; dAs;

InaAs; dTs; InaTs;

dAs; InaTs; dTs;

InaAs; dT-Sup

676
UTRN-
AAAACGACTCCACAA
UTRN
5′
human
dAs; InaAs; dAs; InaAs;

972

dCs; InaGs; dAs;

m02

InaCs; dTs; InaCs;

dCs; InaAs; dCs;

InaAs; dA-Sup

677
UTRN-
CTCCGAGGAAAAACG
UTRN
5′
human
dCs; InaTs; dCs; InaCs;

312

dGs; InaAs; dGs;

m02

InaGs; dAs; InaAs;

dAs; InaAs; dAs;

InaCs; dG-Sup

678
UTRN-
GCTCCGAGGAAAAAC
UTRN
5′
human
dGs; InaCs; dTs; InaCs;

313

dCs; InaGs; dAs;

m02

InaGs; dGs; InaAs;

dAs; InaAs; dAs;

InaAs; dC-Sup

679
UTRN-
CTCGGCGGGAGAAAG
UTRN
5′
human
dCs; InaTs; dCs; InaGs;

975

dGs; InaCs; dGs;

m02

InaGs; dGs; InaAs;

dGs; InaAs; dAs;

InaAs; dG-Sup

680
UTRN-
GAACCGAAATTTT
UTRN
5′
human
dGs; InaAs; dAs; InaCs;

976

dCs; InaGs; dAs;

m02

InaAs; dAs; InaTs;

dTs; InaTs; dT-Sup

681
UTRN-
GAGAAGGGTGCAGAT
UTRN
5′
human
dGs; InaAs; dGs; InaAs;

977

dAs; InaGs; dGs;

m02

InaGs; dTs; InaGs;

dCs; InaAs; dGs;

InaAs; dT-Sup

682
UTRN-
CTCTCCAGATGAGAA
UTRN
5′
human
dCs; InaTs; dCs; InaTs;

978

dCs; InaCs; dAs;

m02

InaGs; dAs; InaTs;

dGs; InaAs; dGs;

InaAs; dA-Sup

683
UTRN-
CAGGGGTCCGCTCTC
UTRN
5′
human
dCs; InaAs; dGs; InaGs;

979

dGs; InaGs; dTs;

m02

InaCs; dCs; InaGs;

dCs; InaTs; dCs;

InaTs; dC-Sup

684
UTRN-
TCCGGGCAGCCAGGG
UTRN
5′
human
dTs; InaCs; dCs; InaGs;

980

dGs; InaGs; dCs;

m02

InaAs; dGs; InaCs;

dCs; InaAs; dGs;

InaGs; dG-Sup

685
UTRN-
GGGGCTCGCCTCCGG
UTRN
5′
human
dGs; InaGs; dGs; InaGs;

981

dCs; InaTs; dCs;

m02

InaGs; dCs; InaCs;

dTs; InaCs; dCs;

InaGs; dG-Sup

686
UTRN-
CCCCCGGGAAGGGGC
UTRN
5′
human
dCs; InaCs; dCs; InaCs;

982

dCs; InaGs; dGs;

m02

InaGs; dAs; InaAs;

dGs; InaGs; dGs;

InaGs; dC-Sup

687
UTRN-
CCCACCCCCCGGGAA
UTRN
5′
human
dCs; InaCs; dCs; InaAs;

983

dCs; InaCs; dCs;

m02

InaCs; dCs; InaCs;

dGs; InaGs; dGs;

InaAs; dA-Sup

688
UTRN-
GCGTTGCCGCCCCCA
UTRN
5′
human
dGs; InaCs; dGs; InaTs;

984
C

dTs; InaGs; dCs;

m02

InaCs; dGs; InaCs;

dCs; InaCs; dCs;

InaCs; dAs; dC-Sup

689
UTRN-
GCTGGGTCGCGCGTT
UTRN
5′
human
dGs; InaCs; dTs; InaGs;

985

dGs; InaGs; dTs;

m02

InaCs; dGs; InaCs;

dGs; InaCs; dGs;

InaTs; dT-Sup

690
UTRN-
GCGCAGGACCGCTGG
UTRN
5′
human
dGs; InaCs; dGs; InaCs;

986

dAs; InaGs; dGs;

m02

InaAs; dCs; InaCs;

dGs; InaCs; dTs;

InaGs; dG-Sup

691
UTRN-
AGGAGGGAGGGTGG
UTRN
5′
human
dAs; InaGs; dGs; InaAs;

987
G

dGs; InaGs; dGs;

m02

InaAs; dGs; InaGs;

dGs; InaTs; dGs;

InaGs; dG-Sup

692
UTRN-
CGCTGGAGGCGGAG
UTRN
5′
human
dCs; InaGs; dCs; InaTs;

988
G

dGs; InaGs; dAs;

m02

InaGs; dGs; InaCs;

dGs; InaGs; dAs;

InaGs; dG-Sup

693
UTRN-
TGGAGCCGAGCGCTG
UTRN
5′
human
dTs; InaGs; dGs; InaAs;

192

dGs; InaCs; dCs;

m02

InaGs; dAs; InaGs;

dCs; InaGs; dCs;

InaTs; dG-Sup

694
UTRN-
CTGCCCCTTTGTTGG
UTRN
5′
human
dCs; InaTs; dGs; InaCs;

303

dCs; InaCs; dCs;

m02

InaTs; dTs; InaTs;

dGs; InaTs; dTs;

InaGs; dG-Sup

695
UTRN-
CTCCCCGCTGCGGGC
UTRN
5′
human
dCs; InaTs; dCs; InaCs;

991

dCs; InaCs; dGs;

m02

InaCs; dTs; InaGs;

dCs; InaGs; dGs;

InaGs; dC-Sup

696
UTRN-
CGGCTCCTCCTCCTC
UTRN
5′
human
dCs; InaGs; dGs; InaCs;

992

dTs; InaCs; dCs;

m02

InaTs; dCs; InaCs;

dTs; InaCs; dCs;

InaTs; dC-Sup

697
UTRN-
GGCTCGCTCCTTCGG
UTRN
5′
human
dGs; InaGs; dCs; InaTs;

993

dCs; InaGs; dCs;

m02

InaTs; dCs; InaCs;

dTs; InaTs; dCs;

InaGs; dG-Sup

698
UTRN-
TTTGTGCGCGAGAGA
UTRN
5′
human
dTs; InaTs; dTs; InaGs;

994

dTs; InaGs; dCs;

m02

InaGs; dCs; InaGs;

dAs; InaGs; dAs;

InaGs; dA-Sup

699
UTRN-
ACGACTCCACAACTT
UTRN
5′
human
dAs; InaCs; dGs; InaAs;

995

dCs; InaTs; dCs;

m02

InaCs; dAs; InaCs;

dAs; InaAs; dCs;

InaTs; dT-Sup

700
UTRN-
GCCCGCTTCCCTGCT
UTRN
5′
human
dGs; InaCs; dCs; InaCs;

997

dGs; InaCs; dTs;

m02

InaTs; dCs; InaCs;

dCs; InaTs; dGs;

InaCs; dT-Sup

701
UTRN-
CGGCCGGCTGCTGCT
UTRN
5′
human
dCs; InaGs; dGs; InaCs;

662

dCs; InaGs; dGs;

m02

InaCs; dTs; InaGs;

dCs; InaTs; dGs;

InaCs; dT-Sup

702
UTRN-
GCGGGAGAAAGCCC
UTRN
5′
human
dGs; InaCs; dGs; InaGs;

999
G

dGs; InaAs; dGs;

m02

InaAs; dAs; InaAs;

dGs; InaCs; dCs;

InaCs; dG-Sup

703
UTRN-
CCTCCTCGCCCCTCG
UTRN
5′
human
dCs; InaCs; dTs; InaCs;

1000

dCs; InaTs; dCs;

m02

InaGs; dCs; InaCs;

dCs; InaCs; dTs;

InaCs; dG-Sup

704
UTRN-
AGAGGCTCCTCCTCG
UTRN
5′
human
dAs; InaGs; dAs; InaGs;

1001

dGs; InaCs; dTs;

m02

InaCs; dCs; InaTs;

dCs; InaCs; dTs;

InaCs; dG-Sup

705
UTRN-
TCGGCTTCTGGAGCC
UTRN
5′
human
dTs; InaCs; dGs; InaGs;

1002

dCs; InaTs; dTs;

m02

InaCs; dTs; InaGs;

dGs; InaAs; dGs;

InaCs; dC-Sup

706
UTRN-
CCGTGATTCCCCAAT
UTRN
5′
human
dCs; InaCs; dGs; InaTs;

1003

dGs; InaAs; dTs;

m02

InaTs; dCs; InaCs;

dCs; InaCs; dAs;

InaAs; dT-Sup

707
UTRN-
AGGGGGGCGCCGCTC
UTRN
5′
human
dAs; InaGs; dGs; InaGs;

1004

dGs; InaGs; dGs;

m02

InaCs; dGs; InaCs;

dCs; InaGs; dCs;

InaTs; dC-Sup

708
UTRN-
AAATGACCCAAAAGA
UTRN
5′
human
dAs; InaAs; dAs; InaTs;

323

dGs; InaAs; dCs;

m02

InaCs; dCs; InaAs;

dAs; InaAs; dAs;

InaGs; dA-Sup

709
UTRN-
GTTTTCCGTTTGCAG
UTRN
5′
human
dGs; InaTs; dTs; InaTs;

328

dTs; InaCs; dCs;

m02

InaGs; dTs; InaTs;

dTs; InaGs; dCs;

InaAs; dG-Sup

710
UTRN-
CCAAACGCTACAGAG
UTRN
5′
human
dCs; InaCs; dAs; InaAs;

334

dAs; InaCs; dGs;

m02

InaCs; dTs; InaAs;

dCs; InaAs; dGs;

InaAs; dG-Sup

711
UTRN-
CAGGCACCAACTTTG
UTRN
5′
human
dCs; InaAs; dGs; InaGs;

1008

dCs; InaAs; dCs;

m02

InaCs; dAs; InaAs;

dCs; InaTs; dTs;

InaTs; dG-Sup

712
UTRN-
CCTGGAAGGGGCGCG
UTRN
5′
human
dCs; InaCs; dTs; InaGs;

1009

dGs; InaAs; dAs;

m02

InaGs; dGs; InaGs;

dGs; InaCs; dGs;

InaCs; dG-Sup

713
UTRN-
CAGTCAAAGCGCAAA
UTRN
5′
human
dCs; InaAs; dGs; InaTs;

345

dCs; InaAs; dAs;

m02

InaAs; dGs; InaCs;

dGs; InaCs; dAs;

InaAs; dA-Sup

714
UTRN-
CCAAAAACAAAACAG
UTRN
5′
human
dCs; InaCs; dAs; InaAs;

1011

dAs; InaAs; dAs;

m02

InaCs; dAs; InaAs;

dAs; InaAs; dCs;

InaAs; dG-Sup

715
UTRN-
TTCCGCCAAAAACAA
UTRN
5′
human
dTs; InaTs; dCs; InaCs;

674

dGs; InaCs; dCs;

m02

InaAs; dAs; InaAs;

dAs; InaAs; dCs;

InaAs; dA-Sup

716
UTRN-
GGAGGAGGGAGGGT
UTRN
5′
human
dGs; InaGs; dAs; InaGs;

1013
G

dGs; InaAs; dGs;

m02

InaGs; dGs; InaAs;

dGs; InaGs; dGs;

InaTs; dG-Sup

717
UTRN-
CGAGCGCTGGAGGCG
UTRN
5′
human
dCs; InaGs; dAs; InaGs;

1014

dCs; InaGs; dCs;

m02

InaTs; dGs; InaGs;

dAs; InaGs; dGs;

InaCs; dG-Sup

718
UTRN-
CCTGCCCCTTTGTTG
UTRN
5′
human
dCs; InaCs; dTs; InaGs;

1015

dCs; InaCs; dCs;

m02

InaCs; dTs; InaTs;

dTs; InaGs; dTs;

InaTs; dG-Sup

719
UTRN-
GGCGGCTCCTCCTCC
UTRN
5′
human
dGs; InaGs; dCs; InaGs;

1016

dGs; InaCs; dTs;

m02

InaCs; dCs; InaTs;

dCs; InaCs; dTs;

InaCs; dC-Sup

Table 10 provides further exemplary non-coding RNA 5′ and 3′ end targeting oligos.

TABLE 10

Oligonucleotides designed to target 5′ and 3′ ends of non-coding RNAs

SEQ
Oligo

Gene
Target

Formatted

ID NO
Name
Base Sequence
Name
Region
Organism
Sequence

720
DINO-1
TAGACACTTCCAGAA
DINO
3′
human
dTs; InaAs; dGs; InaAs;

m02

dCs; InaAs; dCs; InaTs;

dTs; InaCs; dCs; InaAs;

dGs; InaAs; dA-

Sup

721
DINO-2
TTCCAGAATTGTCCT
DINO
3′
human
dTs; InaTs; dCs; InaCs;

m02

dAs; InaGs; dAs; InaAs;

dTs; InaTs; dGs; InaTs;

dCs; InaCs; dT-

Sup

722
DINO-3
CAGAATTGTCCTTTA
DINO
3′
human
dCs; InaAs; dGs; InaAs;

m02

dAs; InaTs; dTs; InaGs;

dTs; InaCs; dCs; InaTs;

dTs; InaTs; dA-

Sup

723
DINO-4
CTGCTGGAACTCGGC
DINO
5′
human
dCs; InaTs; dGs; InaCs;

m02

dTs; InaGs; dGs; InaAs;

dAs; InaCs; dTs; InaCs;

dGs; InaGs; dC-

Sup

724
DINO-5
GGCCAGGCTCAGCTG
DINO
5′
human
dGs; InaGs; dCs; InaCs;

m02

dAs; InaGs; dGs; InaCs;

dTs; InaCs; dAs; InaGs;

dCs; InaTs; dG-

Sup

725
DINO-6
GCAGCCAGGAGCCTG
DINO
5′
human
dGs; InaCs; dAs; InaGs;

m02

dCs; InaCs; dAs; InaGs;

dGs; InaAs; dGs; InaCs;

dCs; InaTs; dG-

Sup

726
DINO-7
ACTCGGCCAGGCTCA
DINO
5′
human
dAs; InaCs; dTs; InaCs;

m02

dGs; InaGs; dCs; InaCs;

dAs; InaGs; dGs;

InaCs; dTs; InaCs; dA-

Sup

727
DINO-8
GCTGGCCTGCTGGAA
DINO
5′
human
dGs; InaCs; dTs; InaGs;

m02

dGs; InaCs; dCs; InaTs;

dGs; InaCs; dTs; InaGs;

dGs; InaAs; dA-

Sup

728
HOTTIP-1
TTTAAATTGTATCGG
HOTTIP
3′
human
dTs; InaTs; dTs; InaAs;

m02

dAs; InaAs; dTs; InaTs;

dGs; InaTs; dAs; InaTs;

dCs; InaGs; dG-

Sup

729
HOTTIP-2
ATTGTATCGGGCAAA
HOTTIP
3′
human
dAs; InaTs; dTs; InaGs;

m02

dTs; InaAs; dTs; InaCs;

dGs; InaGs; dGs; InaCs;

dAs; InaAs; dA-

Sup

730
HOTTIP-3
GATTAAAACAAAAGA
HOTTIP
3′
human
dGs; InaAs; dTs; InaTs;

m02

dAs; InaAs; dAs; InaAs;

dCs; InaAs; dAs; InaAs;

dAs; InaGs; dA-

Sup

731
HOTTIP-4
AAAACAAAAGAAACC
HOTTIP
3′
human
dAs; InaAs; dAs; InaAs;

m02

dCs; InaAs; dAs; InaAs;

dAs; InaGs; dAs; InaAs;

dAs; InaCs; dC-

Sup

732
HOTTIP-5
GGGATAAAGGAAGGG
HOTTIP
5′
human
dGs; InaGs; dGs; InaAs;

m02

dTs; InaAs; dAs; InaAs;

dGs; InaGs; dAs;

InaAs; dGs; InaGs;

dG-Sup

733
HOTTIP-6
CACTGGGATAAAGGA
HOTTIP
5′
human
dCs; InaAs; dCs; InaTs;

m02

dGs; InaGs; dGs; InaAs;

dTs; InaAs; dAs; InaAs;

dGs; InaGs; dA-

Sup

734
HOTTIP-7
GAGCCGCCCGCTTTG
HOTTIP
5′
human
dGs; InaAs; dGs; InaCs;

m02

dCs; InaGs; dCs; InaCs;

dCs; InaGs; dCs; InaTs;

dTs; InaTs; dG-

Sup

735
HOTTIP-8
TCTGGGCCCCACTG
HOTTIP
5′
human
dTs; InaCs; dTs; InaGs;

m02

dGs; InaGs; dCs; InaCs;

dCs; InaCs; dAs; InaCs;

dTs; InaG-Sup

736
NEST-1
CAAAAGGTCTTAGCT
NEST
3′
human
dCs; InaAs; dAs; InaAs;

m02

dAs; InaGs; dGs; InaTs;

dCs; InaTs; dTs; InaAs;

dGs; InaCs; dT-

Sup

737
NEST-2
TAGCTATTATTACTG
NEST
3′
human
dTs; InaAs; dGs; InaCs;

m02

dTs; InaAs; dTs; InaTs;

dAs; InaTs; dTs; InaAs;

dCs; InaTs; dG-

Sup

738
NEST-3
ACTGTTGTTGTTTTA
NEST
3′
human
dAs; InaCs; dTs; InaGs;

m02

dTs; InaTs; dGs; InaTs;

dTs; InaGs; dTs; InaTs;

dTs; InaTs; dA-

Sup

739
NEST-4
ACCTTAGAGGTTGTA
NEST
3′
human
dAs; InaCs; dCs; InaTs;

m02

dTs; InaAs; dGs; InaAs;

dGs; InaGs; dTs; InaTs;

dGs; InaTs; dA-

Sup

740
NEST-5
TACCTGAAATTGCAG
NEST
5′
human
dTs; InaAs; dCs; InaCs;

m02

dTs; InaGs; dAs; InaAs;

dAs; InaTs; dTs; InaGs;

dCs; InaAs; dG-

Sup

741
NEST-6
GTCAGAAAAGCTACC
NEST
5′
human
dGs; InaTs; dCs; InaAs;

m02

dGs; InaAs; dAs; InaAs;

dAs; InaGs; dCs;

InaTs; dAs; InaCs; dC-

Sup

742
NEST-7
CACGCTTGGTGTGCA
NEST
5′
human
dCs; InaAs; dCs; InaGs;

m02

dCs; InaTs; dTs; InaGs;

dGs; InaTs; dGs; InaTs;

dGs; InaCs; dA-

Sup

743
NEST-8
CTGTGAATGTGTGAA
NEST
5′
human
dCs; InaTs; dGs; InaTs;

m02

dGs; InaAs; dAs; InaTs;

dGs; InaTs; dGs; InaTs;

dGs; InaAs; dA-

Sup

744
NEST-9
AACAGGAAGCACCTG
NEST
5′
human
dAs; InaAs; dCs; InaAs;

m02

dGs; InaGs; dAs; InaAs;

dGs; InaCs; dAs;

InaCs; dCs; InaTs; dG-

Sup

TABLE 11

Sequences for FXN-434 and FXN-436

SEQ
Oligo

Gene
Target

Formatted

ID NO
Name
Base Sequence
Name
Region
Organism
Sequence

745
FXN-434
CGCTCCGCCCTCCAGTTT
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs;

m02
TTTTTTAGGAGGCAACA

dCs; InaCs; dGs; InaCs;

CATT

dCs; InaCs; dTs; InaCs;

dCs; InaAs; dG; dT;

dT; dT; dT; dTs; InaTs;

dTs; InaTs; dTs; InaAs;

dGs; InaGs; dAs;

InaGs; dGs; InaCs; dAs;

InaAs; dCs; InaAs; dCs;

InaAs; dTs; InaT-

Sup

746
FXN-436
CGCTCCGCCCTCCAGCC
FXN
5′ and 3′
human
dCs; InaGs; dCs; InaTs;

m02
TTTTTTTTTAGGAGGCA

dCs; InaCs; dGs; InaCs;

ACACATT

dCs; InaCs; dTs; InaCs;

dCs; InaAs; dGs; InaCs;

dC; dT; dT; dT; dT;

dTs; InaTs; dTs; InaTs;

dTs; InaAs; dGs; InaGs;

dAs; InaGs; dGs;

InaCs; dAs; InaAs; dCs;

InaAs; dCs; InaAs; dTs;

InaT-Sup

TABLE 12

Oligonucleotides targeting 5′ and 3′ ends of FXN, APOA1, THRB, HAMP

and NR1H4

Oligo

SEQ ID

Name
Gene
Organism
NO.
Base Sequence
Formatted Sequence

THRB-
THRB
human
747
CTGTTATAAGCTTTT
InaCs; omeUs; InaGs; omeUs; InaTs; omeAs;

67

InaTs; omeAs; InaAs; omeGs; InaCs;

m01

omeUs; InaTs; omeUs; InaT-Sup

THRB-
THRB
human
748
GTTATAAGCTTTTTC
InaGs; omeUs; InaTs; omeAs; InaTs; ome

68

As; InaAs; omeGs; InaCs; omeUs; InaTs;

m01

omeUs; InaTs; omeUs; InaC-Sup

THRB-
THRB
human
749
TATCTGTTATAAGCT
InaTs; omeAs; InaTs; omeCs; InaTs; omeGs;

69

InaTs; omeUs; InaAs; omeUs; InaAs; omeAs;

m01

InaGs; omeCs; InaT-Sup

THRB-
THRB
human
750
AGTAGATGTTTATTT
InaAs; omeGs; InaTs; omeAs; InaGs; omeAs;

70

InaTs; omeGs; InaTs; omeUs; InaTs; omeAs;

m01

InaTs; omeUs; InaT-Sup

THRB-
THRB
human
751
TAGGCAAAGGAATAG
InaTs; omeAs; InaGs; omeGs; InaCs; omeAs;

71

InaAs; omeAs; InaGs; omeGs; InaAs;

m01

omeAs; InaTs; omeAs; InaG-Sup

THRB-
THRB
human
752
GGTAGGCAAAGGAAT
InaGs; omeGs; InaTs; omeAs; InaGs; omeGs;

72

InaCs; omeAs; InaAs; omeAs; InaGs;

m01

omeGs; InaAs; omeAs; InaT-Sup

THRB-
THRB
human
753
GGCAAAGGAATAGTT
InaGs; omeGs; InaCs; omeAs; InaAs; omeAs;

73

InaGs; omeGs; InaAs; omeAs; InaTs;

m01

omeAs; InaGs; omeUs; InaT-Sup

THRB-
THRB
human
754
GAAATGACACCCAGT
InaGs; omeAs; InaAs; omeAs; InaTs; omeGs;

74

InaAs; omeCs; InaAs; omeCs; InaCs;

m01

omeCs; InaAs; omeGs; InaT-Sup

THRB-
THRB
human
755
AAATGACACCCAGTA
InaAs; omeAs; InaAs; omeUs; InaGs; omeAs;

75

InaCs; omeAs; InaCs; omeCs; InaCs;

m01

omeAs; InaGs; omeUs; InaA-Sup

THRB-
THRB
human
756
GGCAATGGAATGAAA
InaGs; omeGs; InaCs; omeAs; InaAs; omeUs;

76

InaGs; omeGs; InaAs; omeAs; InaTs;

m01

omeGs; InaAs; omeAs; InaA-Sup

THRB-
THRB
human
757
CAATGGAATGAAATG
InaCs; omeAs; InaAs; omeUs; InaGs; omeGs;

77

InaAs; omeAs; InaTs; omeGs; InaAs;

m01

omeAs; InaAs; omeUs; InaG-Sup

THRB-
THRB
human
758
ATGGAATGAAATGAC
InaAs; omeUs; InaGs; omeGs; InaAs; omeAs;

78

InaTs; omeGs; InaAs; omeAs; InaAs;

m01

omeUs; InaGs; omeAs; InaC-Sup

THRB-
THRB
human
759
GTTTCAAGTACCCGC
InaGs; omeUs; InaTs; omeUs; InaCs; omeAs;

79

InaAs; omeGs; InaTs; omeAs; InaCs;

m01

omeCs; InaCs; omeGs; InaC-Sup

THRB-
THRB
human
760
GACCGGAGAACGAAA
InaGs; omeAs; InaCs; omeCs; InaGs; omeGs;

80

InaAs; omeGs; InaAs; omeAs; InaCs;

m01

omeGs; InaAs; omeAs; InaA-Sup

THRB-
THRB
human
761
CTTTGGAAGGTGTTT
InaCs; omeUs; InaTs; omeUs; InaGs; omeGs;

81

InaAs; omeAs; InaGs; omeGs; InaTs;

m01

omeGs; InaTs; omeUs; InaT-Sup

THRB-
THRB
human
762
TTTCTTTGGAAGGTG
InaTs; omeUs; InaTs; omeCs; InaTs; omeUs;

82

InaTs; omeGs; InaGs; omeAs; InaAs;

m01

omeGs; InaGs; omeUs; InaG-Sup

THRB-
THRB
human
763
AGTTAATCCCCGCCG
InaAs; omeGs; InaTs; omeUs; InaAs; omeAs;

83

InaTs; omeCs; InaCs; omeCs; InaCs;

m01

omeGs; InaCs; omeCs; InaG-Sup

THRB-
THRB
human
764
TCCTGCAAAATGTCA
InaTs; omeCs; InaCs; omeUs; InaGs; omeCs;

84

InaAs; omeAs; InaAs; omeAs; InaTs;

m01

omeGs; InaTs; omeCs; InaA-Sup

THRB-
THRB
human
765
CCCCGCAGTCTCCAC
InaCs; omeCs; InaCs; omeCs; InaGs; omeCs;

85

InaAs; omeGs; InaTs; omeCs; InaTs;

m01

omeCs; InaCs; omeAs; InaC-Sup

THRB-
THRB
human
766
TCTCCACCCTCCTCC
InaTs; omeCs; InaTs; omeCs; InaCs; omeAs;

86

InaCs; omeCs; InaCs; omeUs; InaCs;

m01

omeCs; InaTs; omeCs; InaC-Sup

THRB-
THRB
human
767
GAGCGCCGGCGACTG
InaGs; omeAs; InaGs; omeCs; InaGs; omeCs;

87

InaCs; omeGs; InaGs; omeCs; InaGs;

m01

omeAs; InaCs; omeUs; InaG-Sup

THRB-
THRB
human
768
AGGATGTGCGCCTTC
InaAs; omeGs; InaGs; omeAs; InaTs; omeGs;

88

InaTs; omeGs; InaCs; omeGs; InaCs;

m01

omeCs; InaTs; omeUs; InaC-Sup

THRB-
THRB
human
769
GGCGCAGCGAGGAA
InaGs; omeGs; InaCs; omeGs; InaCs; omeAs;

89

InaGs; omeCs; InaGs; omeAs; InaGs;

m01

omeGs; InaAs; InaA-Sup

THRB-
THRB
human
770
TTTCACTGACATCTC
InaTs; omeUs; InaTs; omeCs; InaAs; omeCs;

90

InaTs; omeGs; InaAs; omeCs; InaAs;

m01

omeUs; InaCs; omeUs; InaC-Sup

HAMP
HAMP
human
771
GGTGGTCTGAGCCCC
InaGs; omeGs; InaTs; omeGs; InaGs; omeUs;

01

InaCs; omeUs; InaGs; omeAs; InaGs;

m01

omeCs; InaCs; omeCs; InaC-Sup

HAMP
HAMP
human
772
GGGCCTGCCAGGGGA
InaGs; omeGs; InaGs; omeCs; InaCs; omeUs;

02

InaGs; omeCs; InaCs; omeAs; InaGs;

m01

omeGs; InaGs; omeGs; InaA-Sup

HAMP-
HAMP
human
773
ACCGAGTGACAGTCG
InaAs; omeCs; InaCs; omeGs; InaAs; omeGs;

03

InaTs; omeGs; InaAs; omeCs; InaAs;

m01

omeGs; InaTs; omeCs; InaG-Sup

HAMP-
HAMP
human
774
GTCTGGGACCGAGTG
InaGs; omeUs; InaCs; omeUs; InaGs; omeGs;

04

InaGs; omeAs; InaCs; omeCs; InaGs;

m01

omeAs; InaGs; omeUs; InaG-Sup

HAMP-
HAMP
human
775
TGGGACCGAGTGACA
InaTs; omeGs; InaGs; omeGs; InaAs; omeCs;

05

InaCs; omeGs; InaAs; omeGs; InaTs;

m01

omeGs; InaAs; omeCs; InaA-Sup

HAMP-
HAMP
human
776
GCAGAGGTGTGTTCA
InaGs; omeCs; InaAs; omeGs; InaAs; omeGs;

06

InaGs; omeUs; InaGs; omeUs; InaGs;

m01

omeUs; InaTs; omeCs; InaA-Sup

HAMP-
HAMP
human
777
CGGCAGAGGTGTGTT
InaCs; omeGs; InaGs; omeCs; InaAs; omeGs;

07

InaAs; omeGs; InaGs; omeUs; InaGs;

m01

omeUs; InaGs; omeUs; InaT-Sup

HAMP-
HAMP
human
778
GGGCAGACGGGGTCA
InaGs; omeGs; InaGs; omeCs; InaAs; omeGs;

08

InaAs; omeCs; InaGs; omeGs; InaGs;

m01

omeGs; InaTs; omeCs; InaA-Sup

HAMP-
HAMP
human
779
CTCTGGTTTGGAAAA
InaCs; omeUs; InaCs; omeUs; InaGs; omeGs;

09

InaTs; omeUs; InaTs; omeGs; InaGs;

m01

omeAs; InaAs; omeAs; InaA-Sup

HAMP-
HAMP
human
780
CTGGTTTGGAAAACA
InaCs; omeUs; InaGs; omeGs; InaTs; omeUs;

10

InaTs; omeGs; InaGs; omeAs; InaAs;

m01

omeAs; InaAs; omeCs; InaA-Sup

HAMP-
HAMP
human
781
GTTTGGAAAACAAAA
InaGs; omeUs; InaTs; omeUs; InaGs; omeGs;

11

InaAs; omeAs; InaAs; omeAs; InaCs;

m01

omeAs; InaAs; omeAs; InaA-Sup

HAMP-
HAMP
human
782
GGAAAACAAAAGAAC
InaGs; omeGs; InaAs; omeAs; InaAs; omeAs;

12

InaCs; omeAs; InaAs; omeAs; InaAs;

m01

omeGs; InaAs; omeAs; InaC-Sup

HAMP-
HAMP
human
783
GAAAACAAAAGAACC
InaGs; omeAs; InaAs; omeAs; InaAs; omeCs;

13

InaAs; omeAs; InaAs; omeAs; InaGs;

m01

omeAs; InaAs; omeCs; InaC-Sup

HAMP-
HAMP
human
784
TTTGGAAAACAAAAG
InaTs; omeUs; InaTs; omeGs; InaGs; omeAs;

14

InaAs; omeAs; InaAs; omeCs; InaAs;

m01

omeAs; InaAs; omeAs; InaG-Sup

HAMP-
HAMP
human
785
TGGAAAACAAAAGAA
InaTs; omeGs; InaGs; omeAs; InaAs; omeAs;

15

InaAs; omeCs; InaAs; omeAs; InaAs;

m01

omeAs; InaGs; omeAs; InaA-Sup

HAMP-
HAMP
human
786
TCTGGGGCAGCAGGA
InaTs; omeCs; InaTs; omeGs; InaGs; omeGs;

16

InaGs; omeCs; InaAs; omeGs; InaCs;

m01

omeAs; InaGs; omeGs; InaA-Sup

NR1H4-
NR1H4
human
787
ATAGAAAGGAACCTT
InaAs; omeUs; InaAs; omeGs; InaAs; omeAs;

01

InaAs; omeGs; InaGs; omeAs; InaAs;

m01

omeCs; InaCs; omeUs; InaT-Sup

NR1H4-
NR1H4
human
788
TAGAAAGGAACCTTG
InaTs; omeAs; InaGs; omeAs; InaAs; omeAs;

02

InaGs; omeGs; InaAs; omeAs; InaCs;

m01

omeCs; InaTs; omeUs; InaG-Sup

NR1H4-
NR1H4
human
789
ATTAACAATCCTTCC
InaAs; omeUs; InaTs; omeAs; InaAs; omeCs;

03

InaAs; omeAs; InaTs; omeCs; InaCs;

m01

omeUs; InaTs; omeCs; InaC-Sup

NR1H4-
NR1H4
human
790
CTTCCCTGCTAAATG
InaCs; omeUs; InaTs; omeCs; InaCs; omeCs;

04

InaTs; omeGs; InaCs; omeUs; InaAs;

m01

omeAs; InaAs; omeUs; InaG-Sup

NR1H4-
NR1H4
human
791
CCCTGCTAAATGATA
InaCs; omeCs; InaCs; omeUs; InaGs; omeCs;

05

InaTs; omeAs; InaAs; omeAs; InaTs;

m01

omeGs; InaAs; omeUs; InaA-Sup

NR1H4-
NR1H4
human
792
TGATATAAACATAGA
InaTs; omeGs; InaAs; omeUs; InaAs; omeUs;

06

InaAs; omeAs; InaAs; omeCs; InaAs;

m01

omeUs; InaAs; omeGs; InaA-Sup

NR1H4-
NR1H4
human
793
TCCCCGGGACTGAAC
InaTs; omeCs; InaCs; omeCs; InaCs; omeGs;

07

InaGs; omeGs; InaAs; omeCs; InaTs;

m01

omeGs; InaAs; omeAs; InaC-Sup

NR1H4-
NR1H4
human
794
TACAACTTCTTTGAAT
InaTs; omeAs; InaCs; omeAs; InaAs; omeCs;

08

InaTs; omeUs; InaCs; omeUs; InaTs;

m01

omeUs; InaGs; omeAs; InaAs; InaT-Sup

NR1H4-
NR1H4
human
795
TCTATCACTTCCCCG
InaTs; omeCs; InaTs; omeAs; InaTs; omeCs;

09

InaAs; omeCs; InaTs; omeUs; InaCs;

m01

omeCs; InaCs; omeCs; InaG-Sup

NR1H4-
NR1H4
human
796
TCCTGGGCACCCGTA
InaTs; omeCs; InaCs; omeUs; InaGs; omeGs;

10

InaGs; omeCs; InaAs; omeCs; InaCs;

m01

omeCs; InaGs; omeUs; InaA-Sup

NR1H4-
NR1H4
human
797
TTTCTGTACACATCA
InaTs; omeUs; InaTs; omeCs; InaTs; omeGs;

11

InaTs; omeAs; InaCs; omeAs; InaCs;

m01

omeAs; InaTs; omeCs; InaA-Sup

NR1H4-
NR1H4
human
798
ACAGCCACTGAAAAT
InaAs; omeCs; InaAs; omeGs; InaCs; omeCs;

12

InaAs; omeCs; InaTs; omeGs; InaAs;

m01

omeAs; InaAs; omeAs; InaT-Sup

NR1H4-
NR1H4
human
799
TTCACAGCCACTGAA
InaTs; omeUs; InaCs; omeAs; InaCs; omeAs;

13

InaGs; omeCs; InaCs; omeAs; InaCs;

m01

omeUs; InaGs; omeAs; InaA-Sup

NR1H4-
NR1H4
human
800
TAAAGAAATGAGTTT
InaTs; omeAs; InaAs; omeAs; InaGs; omeAs;

14

InaAs; omeAs; InaTs; omeGs; InaAs;

m01

omeGs; InaTs; omeUs; InaT-Sup

NR1H4-
NR1H4
human
801
ACATTTAAAGAAATG
InaAs; omeCs; InaAs; omeUs; InaTs; omeUs;

15

InaAs; omeAs; InaAs; omeGs; InaAs;

m01

omeAs; InaAs; omeUs; InaG-Sup

NR1H4-
NR1H4
human
802
AAGAAATGAGTTTGT
InaAs; omeAs; InaGs; omeAs; InaAs; omeAs;

16

InaTs; omeGs; InaAs; omeGs; InaTs;

m01

omeUs; InaTs; omeGs; InaT-Sup

NR1H4-
NR1H4
human
803
TGCCATTATGTTTGC
InaTs; omeGs; InaCs; omeCs; InaAs; omeUs;

17

InaTs; omeAs; InaTs; omeGs; InaTs;

m01

omeUs; InaTs; omeGs; InaC-Sup

NR1H4-
NR1H4
human
804
TCCTGTTGCCATTAT
InaTs; omeCs; InaCs; omeUs; InaGs; omeUs;

18

InaTs; omeGs; InaCs; omeCs; InaAs;

m01

omeUs; InaTs; omeAs; InaT-Sup

NR1H4-
NR1H4
human
805
GAAAATCCTGTTGCC
InaGs; omeAs; InaAs; omeAs; InaAs; omeUs;

19

InaCs; omeCs; InaTs; omeGs; InaTs;

m01

omeUs; InaGs; omeCs; InaC-Sup

NR1H4-
NR1H4
human
806
CTGTTGCCATTATGT
InaCs; omeUs; InaGs; omeUs; InaTs; omeGs;

20

InaCs; omeCs; InaAs; omeUs; InaTs;

m01

omeAs; InaTs; omeGs; InaT-Sup

NR1H4-
NR1H4
human
807
TAGAATTGAAGTAAC
InaTs; omeAs; InaGs; omeAs; InaAs; omeUs;

21

InaTs; omeGs; InaAs; omeAs; InaGs;

m01

omeUs; InaAs; omeAs; InaC-Sup

NR1H4-
NR1H4
human
808
TGAAGTAACAATCAA
InaTs; omeGs; InaAs; omeAs; InaGs; omeUs;

22

InaAs; omeAs; InaCs; omeAs; InaAs;

m01

omeUs; InaCs; omeAs; InaA-Sup

NR1H4-
NR1H4
human
809
AAGTAACAATCAATT
InaAs; omeAs; InaGs; omeUs; InaAs; omeAs;

23

InaCs; omeAs; InaAs; omeUs; InaCs;

m01

omeAs; InaAs; omeUs; InaT-Sup

NR1H4-
NR1H4
human
810
TCATCAAGATTTCTT
InaTs; omeCs; InaAs; omeUs; InaCs; omeAs;

24

InaAs; omeGs; InaAs; omeUs; InaTs;

m01

omeUs; InaCs; omeUs; InaT-Sup

NR1H4-
NR1H4
human
811
TATCTAGCCCAATAT
InaTs; omeAs; InaTs; omeCs; InaTs; omeAs;

25

InaGs; omeCs; InaCs; omeCs; InaAs;

m01

omeAs; InaTs; omeAs; InaT-Sup

NR1H4-
NR1H4
human
812
TTCTATCTAGCCCAA
InaTs; omeUs; InaCs; omeUs; InaAs; omeUs;

26

InaCs; omeUs; InaAs; omeGs; InaCs;

m01

omeCs; InaCs; omeAs; InaA-Sup

FXN-
FXN
human
813
CTCCGCCCTCCAGTTT
InaCs; omeUs; omeCs; omeCs; InaGs; omeCs;

837

TTATTATTTTGCTTTTT
omeCs; omeCs; InaTs; omeCs; omeCs;

m1000

omeAs; InaGs; omeUs; omeUs; omeUs;

InaTs; omeUs; omeAs; omeUs; InaTs; omeAs;

omeUs; omeUs; InaTs; omeUs; omeGs;

omeCs; InaTs; omeUs; omeUs; omeUs;

InaT-Sup

FXN-
FXN
human
814
CCGCCCTCCAGTTTTT
InaCs; omeCs; omeGs; omeCs; InaCs; omeCs;

838

ATTATTTTGCTTTTT
omeUs; omeCs; InaCs; omeAs; omeGs;

m1000

omeUs; InaTs; omeUs; omeUs; omeUs;

InaAs; omeUs; omeUs; omeAs; InaTs;

omeUs; omeUs; omeUs; InaGs; omeCs;

omeUs; omeUs; InaTs; omeUs; InaT-Sup

FXN-
FXN
human
815
GCCCTCCAGTTTTTAT
InaGs; omeCs; omeCs; omeCs; InaTs;

839

TATTTTGCTTTTT
omeCs; omeCs; omeAs; InaGs; omeUs;

m1000

omeUs; omeUs; InaTs; omeUs; omeAs;

omeUs; InaTs; omeAs; omeUs; omeUs; InaTs;

omeUs; omeGs; omeCs; InaTs; omeUs;

omeUs; omeUs; InaT-Sup

FXN-
FXN
human
816
CGCTCCGCCCTCCAGT
InaCs; omeGs; omeCs; omeUs; InaCs;

840

TTTTATTATTTTGCTTT
omeCs; omeGs; omeCs; InaCs; omeCs;

m1000

omeUs; omeCs; InaCs; omeAs; omeGs;

omeUs; InaTs; omeUs; omeUs; omeUs; InaAs;

omeUs; omeUs; omeAs; InaTs; omeUs;

omeUs; omeUs; InaGs; omeCs; omeUs;

omeUs; InaT-Sup

FXN-
FXN
human
817
CGCTCCGCCCTCCAGT
InaCs; omeGs; omeCs; omeUs; InaCs;

841

TTTTATTATTTTGCT
omeCs; omeGs; omeCs; InaCs; omeCs;

m1000

omeUs; omeCs; InaCs; omeAs; omeGs;

omeUs; InaTs; omeUs; omeUs; omeUs; InaAs;

omeUs; omeUs; omeAs; InaTs; omeUs;

omeUs; omeUs; InaGs; omeCs; InaT-Sup

FXN-
FXN
human
818
CGCTCCGCCCTCCAGT
InaCs; omeGs; omeCs; omeUs; InaCs;

842

TTTTATTATTTTG
omeCs; omeGs; omeCs; InaCs; omeCs;

m1000

omeUs; omeCs; InaCs; omeAs; omeGs;

omeUs; InaTs; omeUs; omeUs; omeUs; InaAs;

omeUs; omeUs; omeAs; InaTs; omeUs;

omeUs; omeUs; InaG-Sup

FXN-
FXN
human
819
CTCCGCCCTCCAGTTT
InaCs; omeUs; omeCs; omeCs; InaGs;

843

TTATTATTTTGCTTT
omeCs; omeCs; omeCs; InaTs; omeCs; omeCs;

m1000

omeAs; InaGs; omeUs; omeUs; omeUs;

InaTs; omeUs; omeAs; omeUs; InaTs;

omeAs; omeUs; omeUs; InaTs; omeUs;

omeGs; omeCs; InaTs; omeUs; InaT-Sup

FXN-
FXN
human
820
CCGCCCTCCAGTTTTT
InaCs; omeCs; omeGs; InaCs; omeCs;

844

ATTATTTTGCT
omeCs; InaTs; omeCs; omeCs; InaAs; omeGs;

m1000

omeUs; InaTs; omeUs; omeUs; InaTs;

omeAs; omeUs; InaTs; omeAs; omeUs; InaTs;

omeUs; omeUs; InaGs; omeCs; InaT-Sup

FXN-
FXN
human
821
GCCCTCCAGTTTTTAT
InaGs; omeCs; omeCs; InaCs; omeUs;

845

TATTTTGCT
omeCs; InaCs; omeAs; omeGs; InaTs; omeUs;

m1000

omeUs; InaTs; omeUs; omeAs; InaTs;

omeUs; omeAs; InaTs; omeUs; omeUs; InaTs;

omeGs; omeCs; InaT-Sup

FXN-
FXN
human
822
CCCTCCAGTTTTTATT
InaCs; omeCs; omeCs; InaTs; omeCs; omeCs;

846

ATTTTGC
InaAs; omeGs; omeUs; InaTs; omeUs;

m1000

omeUs; InaTs; omeAs; omeUs; InaTs; omeAs;

omeUs; InaTs; omeUs; omeUs; InaGs;

InaC-Sup

FXN-
FXN
human
823
CCTCCAGTTTTTATTAT
InaCs; omeCs; omeUs; InaCs; omeCs; omeAs;

847

TTTG
InaGs; omeUs; omeUs; InaTs; omeUs;

m1000

omeUs; InaAs; omeUs; omeUs; InaAs;

omeUs; omeUs; InaTs; omeUs; InaG-Sup

FXN-
FXN
human
824
GCTCCGCCCTCCAGAT
InaGs; omeCs; omeUs; omeCs; InaCs; omeGs;

848

TATTTTGCTTTTT
omeCs; omeCs; InaCs; omeUs; omeCs;

m1000

omeCs; InaAs; omeGs; omeAs; omeUs;

InaTs; omeAs; omeUs; omeUs; InaTs;

omeUs; omeGs; omeCs; InaTs; omeUs;

omeUs; omeUs; InaT-Sup

FXN-
FXN
human
825
TCCGCCCTCCAGATTA
InaTs; omeCs; omeCs; InaGs; omeCs; omeCs;

849

TTTTGCTTTTT
InaCs; omeUs; omeCs; InaCs; omeAs;

m1000

omeGs; InaAs; omeUs; omeUs; InaAs;

omeUs; omeUs; InaTs; omeUs; omeGs;

InaCs; omeUs; omeUs; InaTs; omeUs; InaT-

Sup

FXN-
FXN
human
826
CGCCCTCCAGATTATT
InaCs; omeGs; omeCs; InaCs; omeCs; omeUs;

850

TTGCTTTTT
InaCs; omeCs; omeAs; InaGs; omeAs;

m1000

omeUs; InaTs; omeAs; omeUs; InaTs;

omeUs; omeUs; InaGs; omeCs; omeUs; InaTs;

omeUs; omeUs; InaT-Sup

FXN-
FXN
human
827
CCCTCCAGATTATTTT
InaCs; omeCs; omeCs; InaTs; omeCs; omeCs;

851

GCTTTTT
InaAs; omeGs; omeAs; InaTs; omeUs;

m1000

omeAs; InaTs; omeUs; omeUs; InaTs;

omeGs; omeCs; InaTs; omeUs; omeUs; InaTs;

InaT-Sup

FXN-
FXN
human
828
GCTCCGCCCTCCAGAT
InaGs; omeCs; omeUs; InaCs; omeCs; omeGs;

852

TATTTTGCTTT
InaCs; omeCs; omeCs; InaTs; omeCs;

m1000

omeCs; InaAs; omeGs; omeAs; InaTs; omeUs;

omeAs; InaTs; omeUs; omeUs; InaTs;

omeGs; omeCs; InaTs; omeUs; InaT-Sup

FXN-
FXN
human
829
GCTCCGCCCTCCAGAT
InaGs; omeCs; omeUs; InaCs; omeCs; omeGs;

853

TATTTTGCT
InaCs; omeCs; omeCs; InaTs; omeCs;

m1000

omeCs; InaAs; omeGs; omeAs; InaTs; omeUs;

omeAs; InaTs; omeUs; omeUs; InaTs;

omeGs; omeCs; InaT-Sup

FXN-
FXN
human
830
GCTCCGCCCTCCAGAT
InaGs; omeCs; omeUs; InaCs; omeCs; omeGs;

854

TATTTTG
InaCs; omeCs; omeCs; InaTs; omeCs;

m1000

omeCs; InaAs; omeGs; omeAs; InaTs; omeUs;

omeAs; InaTs; omeUs; omeUs; InaTs;

InaG-Sup

FXN-
FXN
human
831
TCCGCCCTCCAGATTA
InaTs; omeCs; omeCs; InaGs; omeCs; omeCs;

855

TTTTGCTTT
InaCs; omeUs; omeCs; InaCs; omeAs;

m1000

omeGs; InaAs; omeUs; omeUs; InaAs; omeUs;

omeUs; InaTs; omeUs; omeGs; InaCs;

omeUs; omeUs; InaT-Sup

FXN-
FXN
human
832
CGCCCTCCAGATTATT
InaCs; omeGs; omeCs; InaCs; omeCs; omeUs;

856

TTGCT
InaCs; omeCs; omeAs; InaGs; omeAs;

m1000

omeUs; InaTs; omeAs; omeUs; InaTs; omeUs;

omeUs; InaGs; omeCs; InaT-Sup

FXN-
FXN
human
833
GCCCTCCAGATTATTT
InaGs; omeCs; InaCs; omeCs; InaTs; omeCs;

857

TGC
InaCs; omeAs; InaGs; omeAs; InaTs; omeUs;

m01

InaAs; omeUs; InaTs; omeUs; InaTs;

omeGs; InaC-Sup

FXN-
FXN
human
834
CCCTCCAGATTATTTT
InaCs; omeCs; InaCs; omeUs; InaCs; omeCs;

858

G
InaAs; omeGs; InaAs; omeUs; InaTs; omeAs;

m01

InaTs; omeUs; InaTs; omeUs; InaG-

Sup

FXN-
FXN
human
835
CTCCAGATTATTTTG
InaCs; omeUs; InaCs; omeCs; InaAs; omeGs;

859

InaAs; omeUs; InaTs; omeAs; InaTs; omeUs;

m01

InaTs; omeUs; InaG-Sup

FXN-
FXN
human
836
CGCTCCGCCCTCCAGT
dCs; InaGs; dCs; InaTs; dCs; InaCs; dGs; InaCs;

461

TTTTATTATTTTGCTTT
dCs; InaCs; dTs; InaCs; dCs; InaAs; dGs; InaTs;

m02

TT
dTs; InaTs; dTs; InaTs; dAs; InaTs; dTs;

InaAs; dTs; InaTs; dTs; InaTs; dGs; InaCs;

dTs; InaTs; dTs; InaTs; dT-Sup

Apoa1_
APOA1
mouse
837
AGTTCAAGGATCAGC
InaAs; dGs; dTs; InaTs; dCs; dAs; InaAs; dGs;

mus-

CATTTTGGAAAGG
dGs; InaAs; dTs; dCs; InaAs; dGs; dCs; InaCs;

77

dAs; dTs; InaTs; dTs; dTs; InaGs; dGs; dAs;

m1000

InaAs; dAs; dGs; InaG-Sup

Apoa1_
APOA1
mouse
838
TCAAGGATCAGCCATT
InaTs; dCs; dAs; InaAs; dGs; dGs; InaAs; dTs;

mus-

TTGGAAAGG
dCs; InaAs; dGs; dCs; InaCs; dAs; dTs; InaTs;

78

dTs; dTs; InaGs; dGs; dAs; InaAs; dAs; dGs;

m1000

InaG-Sup

Apoa1_
APOA1
mouse
839
AAGGATCAGCCATTTT
InaAs; dAs; dGs; InaGs; dAs; dTs; InaCs; dAs;

mus-

GGAAAGG
dGs; InaCs; dCs; dAs; InaTs; dTs; dTs; InaTs;

79

dGs; dGs; InaAs; dAs; dAs; InaGs; InaG-

m1000

Sup

Apoa1_
APOA1
mouse
840
GGATCAGCCATTTTG
InaGs; dGs; dAs; InaTs; dCs; dAs; InaGs; dCs;

mus-

GAAAGG
dCs; InaAs; dTs; dTs; InaTs; dTs; dGs; InaGs;

80

dAs; dAs; InaAs; dGs; InaG-Sup

m1000

Apoa1_
APOA1
mouse
841
AGTTCAAGGATCAGC
InaAs; dGs; dTs; InaTs; dCs; dAs; InaAs; dGs;

mus-

CATTTTGGAA
dGs; InaAs; dTs; dCs; InaAs; dGs; dCs; InaCs;

81

dAs; dTs; InaTs; dTs; dTs; InaGs; dGs; dAs;

m1000

InaA-Sup

Apoa1_
APOA1
mouse
842
AGTTCAAGGATCAGC
InaAs; dGs; dTs; InaTs; dCs; dAs; InaAs; dGs;

mus-

CATTTTGG
dGs; InaAs; dTs; dCs; InaAs; dGs; dCs; InaCs;

82

dAs; dTs; InaTs; dTs; dTs; InaGs; InaG-

m1000

Sup

Apoa1_
APOA1
mouse
843
GTTCAAGGATCAGCC
InaGs; dTs; dTs; InaCs; dAs; dAs; InaGs; dGs;

mus-

ATTTTGGAAAGG
dAs; InaTs; dCs; dAs; InaGs; dCs; dCs; InaAs;

83

dTs; dTs; InaTs; dTs; dGs; InaGs; dAs; dAs;

m1000

InaAs; dGs; InaG-Sup

Apoa1_
APOA1
mouse
844
TCAAGGATCAGCCATT
InaTs; dCs; dAs; InaAs; dGs; dGs; InaAs; dTs;

mus-

TTGGAAA
dCs; InaAs; dGs; dCs; InaCs; dAs; dTs; InaTs;

84

dTs; dTs; InaGs; dGs; dAs; InaAs; InaA-

m1000

Sup

Apoa1_
APOA1
mouse
845
AAGGATCAGCCATTTT
InaAs; dAs; dGs; InaGs; dAs; dTs; InaCs; dAs;

mus-

GGAAA
dGs; InaCs; dCs; dAs; InaTs; dTs; dTs; InaTs;

85

dGs; dGs; InaAs; dAs; InaA-Sup

m1000

Apoa1_
APOA1
mouse
846
AGGATCAGCCATTTTG
InaAs; dGs; InaGs; dAs; InaTs; dCs; InaAs; dGs;

mus-

GAA
InaCs; dCs; InaAs; dTs; InaTs; dTs; InaTs;

86

dGs; InaGs; dAs; InaA-Sup

m12

Apoa1_
APOA1
mouse
847
GGATCAGCCATTTTG
InaGs; dGs; InaAs; dTs; InaCs; dAs; InaGs;

mus-

GA
dCs; InaCs; dAs; InaTs; dTs; InaTs; dTs; InaGs;

87

dGs; InaA-Sup

m12

Apoa1_
APOA1
mouse
848
CTCCGACAGTCTGCCA
InaCs; dTs; dCs; InaCs; dGs; dAs; InaCs; dAs;

mus-

TTTTGGAAAGGT
dGs; InaTs; dCs; dTs; InaGs; dCs; dCs; InaAs;

88

dTs; dTs; InaTs; dTs; dGs; InaGs; dAs; dAs;

m1000

InaAs; dGs; dGs; InaT-Sup

Apoa1_
APOA1
mouse
849
CGACAGTCTGCCATTT
InaCs; dGs; dAs; InaCs; dAs; dGs; InaTs; dCs;

mus-

TGGAAAGGT
dTs; InaGs; dCs; dCs; InaAs; dTs; dTs; InaTs;

89

dTs; dGs; InaGs; dAs; dAs; InaAs; dGs; dGs;

m1000

InaT-Sup

Apoa1_
APOA1
mouse
850
ACAGTCTGCCATTTTG
InaAs; dCs; dAs; InaGs; dTs; dCs; InaTs; dGs;

mus-

GAAAGGT
dCs; InaCs; dAs; dTs; InaTs; dTs; dTs; InaGs;

90

dGs; dAs; InaAs; dAs; dGs; InaGs; InaT-

m1000

Sup

Apoa1_
APOA1
mouse
851
CTCCGACAGTCTGCCA
InaCs; dTs; dCs; InaCs; dGs; dAs; InaCs; dAs;

mus-

TTTTGGAAA
dGs; InaTs; dCs; dTs; InaGs; dCs; dCs; InaAs;

91

dTs; dTs; InaTs; dTs; dGs; InaGs; dAs; dAs;

m1000

InaA-Sup

Apoa1_
APOA1
mouse
852
CTCCGACAGTCTGCCA
InaCs; dTs; dCs; InaCs; dGs; dAs; InaCs; dAs;

mus-

TTTTGGA
dGs; InaTs; dCs; dTs; InaGs; dCs; dCs; InaAs;

92

dTs; dTs; InaTs; dTs; dGs; InaGs; InaA-

m1000

Sup

Apoa1_
APOA1
mouse
853
CTCCGACAGTCTGCCA
InaCs; dTs; dCs; InaCs; dGs; dAs; InaCs; dAs;

mus-

TTTTG
dGs; InaTs; dCs; dTs; InaGs; dCs; dCs; InaAs;

93

dTs; dTs; InaTs; dTs; InaG-Sup

m1000

Apoa1_
APOA1
mouse
854
CTCCGACAGTCTGCCA
InaCs; dTs; dCs; InaCs; dGs; dAs; InaCs; dAs;

mus-

TTTTGGAAAGG
dGs; InaTs; dCs; dTs; InaGs; dCs; dCs; InaAs;

94

dTs; dTs; InaTs; dTs; dGs; InaGs; dAs; dAs;

m1000

InaAs; dGs; InaG-Sup

Apoa1_
APOA1
mouse
855
CCGACAGTCTGCCATT
InaCs; dCs; dGs; InaAs; dCs; dAs; InaGs; dTs;

mus-

TTGGAAA
dCs; InaTs; dGs; dCs; InaCs; dAs; dTs; InaTs;

95

dTs; dTs; InaGs; dGs; dAs; InaAs; InaA-

m1000

Sup

Apoa1_
APOA1
mouse
856
GACAGTCTGCCATTTT
InaGs; dAs; InaCs; dAs; InaGs; dTs; InaCs; dTs;

mus-

GGA
InaGs; dCs; InaCs; dAs; InaTs; dTs; InaTs;

96

dTs; InaGs; dGs; InaA-Sup

m12

Apoa1_
APOA1
mouse
857
ACAGTCTGCCATTTTG
InaAs; dCs; InaAs; dGs; InaTs; dCs; InaTs; dGs;

mus-

G
InaCs; dCs; InaAs; dTs; InaTs; dTs; InaTs;

97

dGs; InaG-Sup

m12

Apoa1_
APOA1
mouse
858
CGGAGCTCTCCGACA
InaCs; dGs; dGs; InaAs; dGs; dCs; InaTs; dCs;

mus-

CATTTTGGAAAGGTT
dTs; InaCs; dCs; dGs; InaAs; dCs; dAs; InaCs;

98

dAs; dTs; InaTs; dTs; dTs; InaGs; dGs; dAs;

m1000

InaAs; dAs; dGs; InaGs; dTs; InaT-Sup

Apoa1_
APOA1
mouse
859
GGAGCTCTCCGACAC
InaGs; dGs; dAs; InaGs; dCs; dTs; InaCs; dTs;

mus-

ATTTTGGAAAGGTT
dCs; InaCs; dGs; dAs; InaCs; dAs; dCs; InaAs;

99

dTs; dTs; InaTs; dTs; dGs; InaGs; dAs; dAs;

m1000

InaAs; dGs; dGs; InaTs; InaT-Sup

Apoa1_
APOA1
mouse
860
AGCTCTCCGACACATT
InaAs; dGs; dCs; InaTs; dCs; dTs; InaCs; dCs;

mus-

TTGGAAAGG
dGs; InaAs; dCs; dAs; InaCs; dAs; dTs; InaTs;

100

dTs; dTs; InaGs; dGs; dAs; InaAs; dAs; dGs;

m1000

InaG-Sup

Apoa1_
APOA1
mouse
861
CTCTCCGACACATTTT
InaCs; dTs; dCs; InaTs; dCs; dCs; InaGs; dAs;

mus-

GGAAA
dCs; InaAs; dCs; dAs; InaTs; dTs; dTs; InaTs;

101

dGs; dGs; InaAs; dAs; InaA-Sup

m1000

Apoa1_
APOA1
mouse
862
TCTCCGACACATTTTG
InaTs; dCs; InaTs; dCs; InaCs; dGs; InaAs; dCs;

mus-

GAA
InaAs; dCs; InaAs; dTs; InaTs; dTs; InaTs;

102

dGs; InaGs; dAs; InaA-Sup

m12

Apoa1_
APOA1
mouse
863
CTCCGACACATTTTGG
InaCs; dTs; InaCs; dCs; InaGs; dAs; InaCs; dAs;

mus-

A
InaCs; dAs; InaTs; dTs; InaTs; dTs; InaGs;

103

dGs; InaA-Sup

m12

Apoa1_
APOA1
mouse
864
TCCGACACATTTTGG
InaTs; dCs; InaCs; dGs; InaAs; dCs; InaAs; dCs;

mus-

InaAs; dTs; InaTs; dTs; InaTs; dGs; InaG-

104

Sup

m12

TABLE 13

Other exemplary FXN oligos

SEQ ID

Oligo Name
Gene
Organism
NO.
Base Sequence
Formatted Sequence

FXN-375
FXN
human
865
CGCTCCGCCCTCCAG
dCs; InaGs; dCs; InaTs; dCs; InaCs;

dGs; InaCs; dCs; InaCs; dTs; InaCs;

dCs; InaAs; dG-Sup

FXN-390
FXN
human
866
ATTATTTTGCTTTTT
dAs; InaTs; dTs; InaAs; dTs; InaTs;

dTs; InaTs; dGs; InaCs; dTs; InaTs;

dTs; InaTs; dT-Sup

TABLE 14

Oligonucleotide modifications

Symbol
Feature Description

bio
5′ biotin

dAs
DNA w/3′ thiophosphate

dCs
DNA w/3′ thiophosphate

dGs
DNA w/3′ thiophosphate

dTs
DNA w/3′ thiophosphate

dG
DNA

enaAs
ENA w/3′ thiophosphate

enaCs
ENA w/3′ thiophosphate

enaGs
ENA w/3′ thiophosphate

enaTs
ENA w/3′ thiophosphate

fluAs
2′-fluoro w/3′ thiophosphate

fluCs
2′-fluoro w/3′ thiophosphate

fluGs
2′-fluoro w/3′ thiophosphate

fluUs
2′-fluoro w/3′ thiophosphate

lnaAs
LNA w/3′ thiophosphate

lnaCs
LNA w/3′ thiophosphate

lnaGs
LNA w/3′ thiophosphate

lnaTs
LNA w/3′ thiophosphate

omeAs
2′-OMe w/3′ thiophosphate

omeCs
2′-OMe w/3′ thiophosphate

omeGs
2′-OMe w/3′ thiophosphate

omeTs
2′-OMe w/3′ thiophosphate

lnaAs-Sup
LNA w/3′ thiophosphate at 3′ terminus

lnaCs-Sup
LNA w/3′ thiophosphate at 3′ terminus

lnaGs-Sup
LNA w/3′ thiophosphate at 3′ terminus

lnaTs-Sup
LNA w/3′ thiophosphate at 3′ terminus

lnaA-Sup
LNA w/3′ OH at 3′ terminus

lnaC-Sup
LNA w/3′ OH at 3′ terminus

lnaG-Sup
LNA w/3′ OH at 3′ terminus

lnaT-Sup
LNA w/3′ OH at 3′ terminus

omeA-Sup
2′-OMe w/3′ OH at 3′ terminus

omeC-Sup
2′-OMe w/3′ OH at 3′ terminus

omeG-Sup
2′-OMe w/3′ OH at 3′ terminus

omeU-Sup
2′-OMe w/3′ OH at 3′ terminus

dAs-Sup
DNA w/3′ thiophosphate at 3′ terminus

dCs-Sup
DNA w/3′ thiophosphate at 3′ terminus

dGs-Sup
DNA w/3′ thiophosphate at 3′ terminus

dTs-Sup
DNA w/3′ thiophosphate at 3′ terminus

dA-Sup
DNA w/3′ OH at 3′ terminus

dC-Sup
DNA w/3′ OH at 3′ terminus

dG-Sup
DNA w/3′ OH at 3′ terminus

dT-Sup
DNA w/3′ OH at 3′ terminus

The suffix “Sup” in Table 14 indicates that a 3′ end nucleotide may, for synthesis purposes, be conjugated to a solid support. It should be appreciated that in general when conjugated to a solid support for synthesis, the synthesized oligonucleotide is released such that the solid support is not part of the final oligonucleotide product.

Oligos targeting 3′ and 5′ ends of RNAs, as well as pseudocircularization oligos, can upregulate gene expression.

While several embodiments of the present disclosure have been described and illustrated herein, those of ordinary skill in the art will readily envision a variety of other means and/or structures for performing the functions and/or obtaining the results and/or one or more of the advantages described herein, and each of such variations and/or modifications is deemed to be within the scope of the present disclosure. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and/or configurations will depend upon the specific application or applications for which the teachings of the present disclosure is/are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the disclosure described herein. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and equivalents thereto, the disclosure may be practiced otherwise than as specifically described and claimed. The present disclosure is directed to each individual feature, system, article, material, and/or method described herein. In addition, any combination of two or more such features, systems, articles, materials, and/or methods, if such features, systems, articles, materials, and/or methods are not mutually inconsistent, is included within the scope of the present disclosure.

The indefinite articles “a” and “an,” as used herein in the specification and in the claims, unless clearly indicated to the contrary, should be understood to mean “at least one.”

The phrase “and/or,” as used herein in the specification and in the claims, should be understood to mean “either or both” of the elements so conjoined, e.g., elements that are conjunctively present in some cases and disjunctively present in other cases. Other elements may optionally be present other than the elements specifically identified by the “and/or” clause, whether related or unrelated to those elements specifically identified unless clearly indicated to the contrary. Thus, as a non-limiting example, a reference to “A and/or B,” when used in conjunction with open-ended language such as “comprising” can refer, in one embodiment, to A without B (optionally including elements other than B); in another embodiment, to B without A (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.

As used herein in the specification and in the claims, “or” should be understood to have the same meaning as “and/or” as defined above. For example, when separating items in a list, “or” or “and/or” shall be interpreted as being inclusive, e.g., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as “only one of” or “exactly one of,” or, when used in the claims, “consisting of,” will refer to the inclusion of exactly one element of a number or list of elements. In general, the term “or” as used herein shall only be interpreted as indicating exclusive alternatives (e.g. “one or the other but not both”) when preceded by terms of exclusivity, such as “either,” “one of,” “only one of,” or “exactly one of.” “Consisting essentially of,” when used in the claims, shall have its ordinary meaning as used in the field of patent law.

As used herein in the specification and in the claims, the phrase “at least one,” in reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase “at least one” refers, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, “at least one of A and B” (or, equivalently, “at least one of A or B,” or, equivalently “at least one of A and/or B”) can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.

In the claims, as well as in the specification above, all transitional phrases such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” and the like are to be understood to be open-ended, e.g., to mean including but not limited to. Only the transitional phrases “consisting of” and “consisting essentially of” shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03.

Use of ordinal terms such as “first,” “second,” “third,” etc., in the claims to modify a claim element does not by itself connote any priority, precedence, or order of one claim element over another or the temporal order in which acts of a method are performed, but are used merely as labels to distinguish one claim element having a certain name from another element having a same name (but for use of the ordinal term) to distinguish the claim elements.

Number	Date	Country
62288128	Jan 2016	US
62288154	Jan 2016	US
62246290	Oct 2015	US
62246305	Oct 2015	US

NANOPARTICLE FORMULATIONS FOR DELIVERY OF NUCLEIC ACID COMPLEXES

Information

Publication Number

Date Filed

Date Published

Inventors

Original Assignees

CPC

International Classifications

Abstract

Description

Claims

CROSS-REFERENCE TO RELATED APPLICATIONS

PCT Information

Provisional Applications (4)