COMPLEMENT COMPONENT C3 IRNA COMPOSITIONS AND METHODS OF USE THEREOF

Abstract
The present invention relates to RNAi agents, e.g., double stranded RNA (dsRNA) agents, targeting the complement component C3 gene (C3). The invention also relates to methods of using such RNAi agents to inhibit expression of a C3 gene and to methods of preventing and treating a C3-associated disorder, e.g., cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy.
Description
SEQUENCE LISTING

The instant application contains a Sequence Listing which has been filed electronically in XML file format and is hereby incorporated by reference in its entirety. Said XML copy, created on Mar. 1, 2024, is named 121301_10503_SL.xml and is 28,525,134 bytes in size.


BACKGROUND OF THE INVENTION

Complement was first discovered in the 1890s when it was found to aid or “complement” the killing of bacteria by heat-stable antibodies present in normal serum (Walport, M. J. (2001) N Engl J Med. 344:1058). The complement system consists of more than 30 proteins that are either present as soluble proteins in the blood or are present as membrane-associated proteins. Activation of complement leads to a sequential cascade of enzymatic reactions, known as complement activation pathways resulting in the formation of the potent anaphylatoxins C3a and C5a that elicit a plethora of physiological responses that range from chemoattraction to apoptosis. Initially, complement was thought to play a major role in innate immunity where a robust and rapid response is mounted against invading pathogens. However, recently it is becoming increasingly evident that complement also plays an important role in adaptive immunity involving T and B cells that help in elimination of pathogens (Dunkelberger J R and Song W C. (2010) Cell Res. 20:34; Molina H, et al. (1996) Proc Natl Acad Sci USA. 93:3357), in maintaining immunologic memory preventing pathogenic re-invasion, and is involved in numerous human pathological states (Qu, H, et al. (2009) Mol Immunol. 47:185; Wagner, E. and Frank M M. (2010) Nat Rev Drug Discov. 9:43).


Complement activation is known to occur through three different pathways: alternate, classical and lectin (FIG. 1) involving proteins that mostly exist as inactive zymogens that are then sequentially cleaved and activated.


The classical pathway is often activated by antibody-antigen complexes or by the C-reactive protein (CRP), both of which interact with complement component C1q. In addition, the classical pathway can be activated by phosphatidyl serine present in apoptotic bodies in the absence of immune complexes.


The lectin pathway is initiated by the mannose-binding lectins (MBL) that bind to complex carbohydrate residues on the surface of pathogens. The activation of the classical pathway or the lectin pathway leads to activation of the (C4b2b) C3 convertase.


The alternate pathway is activated by the binding of C3b, which is spontaneously generated by the hydrolysis of C3, on targeted surfaces. This surface-bound C3b is then recognized by factor B, forming the complex C3bB. The C3bB complex, in turn, is cleaved by factor D to yield the active form of the C3 convertase of the AP (C3bBb). Both types of C3 convertases will cleave C3, forming C3b. C3b then either binds to more factor B, enhancing the complement activation through the AP (the so-called alternative or amplification loop), or leads to the formation of the active C5 convertase (C3bBbC3b or C4bC2bC3b), which cleaves C5 and triggers the late events that result in the formation of the membrane attack complex (MAC) (C5b-9).


Inappropriate activation of the complement system is responsible for propagating and/or initiating pathology in many different diseases, including, for example, paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), neuromyelitis optica (NMO), multifocal motor neuropathy (MMN), myasthenia gravis (MG), C3 glomerulonephritis, systemic lupus erythmatosis, rheumatoid arthritis, ischemia-reperfusion injuries and neurodegenerative diseases.


There are limited therapies available for the treatment of complement component C3-associated diseases which require time-consuming and invasive administration at a high cost. Accordingly, there is a need in the art for alternative therapies and combination therapies for subjects having a complement component C3-associated disease.


SUMMARY OF THE INVENTION

The present invention provides iRNA compositions which affect the RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of a gene encoding complement component C3. The complement component C3 may be within a cell, e.g., a cell within a subject, such as a human subject.


In an aspect, the invention provides a double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C3 in a cell, wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 0, 1, 2, or 3 nucleotides from the nucleotide sequence of SEQ ID NO:1 and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 1, 2, or 3 nucleotides from the nucleotide sequence of SEQ ID NO:5.


In another aspect, the present invention provides a double stranded ribonucleic acid (dsRNA) for inhibiting expression of complement component C3 in a cell, wherein said dsRNA comprises a sense strand and an antisense strand forming a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding complement component C3, and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 0, 1, 2, or 3 nucleotides from any one of the antisense nucleotide sequences in any one of Tables 2-7, 15, 18, 20-23, 30, and 31.


In one aspect, the present invention provides a double stranded ribonucleic acid (dsRNA) for inhibiting expression of complement component C3 in a cell, wherein said dsRNA comprises a sense strand and an antisense strand forming a double stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 0, 1, 2, or 3 nucleotides from any one of the nucleotide sequence of nucleotides 475-497, 487-509, 490-512, 491-513, 705-727, 809-831, 813-835, 1147-1169, 1437-1459, 1439-1461, 1447-1469, 2596-2618, 2634-2656, 3012-3034, 3334-3356, 3611-3633, 3614-3636, 3622-3655, 3809-3831, 3846-3868, 3847-3869, 3920-3942, 4047-4069, 4061-4083, 4156-4178, 4157-4177, 4162-4184, 4178-4200, 4226-4248, 4369-4391, 4392-4414, 4521-4543, 4522-4544, 4523-4545, 5012-5034 of the nucleotide sequence of SEQ ID NO:1, and the antisense strand comprises at least 19 contiguous nucleotides from the corresponding nucleotide sequence of SEQ ID NO:5.


In one embodiment, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 0, 1, 2, or 3 nucleotides from any one of the nucleotide sequence of nucleotides 705-727, 809-831, or 634-2656 of SEQ ID NO:1. In another embodiment, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 0, 1, 2, or 3 nucleotides from the nucleotide sequence of nucleotides 634-2656 of SEQ ID NO:1.


In one embodiment, the antisense strand comprises at least 15 contiguous nucleotides differing by nor more than 0, 1, 2, or 3 nucleotides from any one of the antisense strand nucleotide sequences of a duplex selected from the group consisting of AD-565541.2, AD-564742, AD-567304, AD-568978, AD-569164, AD-569272.2, AD-569765.2, AD-564730.2, AD-567315, AD-564745.2, AD-571715.2, AD-570714, AD-571826, AD-572041.2, AD-572039.2, AD-572387, AD-568586.2, AD-566837.2, AD-566444.2, AD-567700.2, AD-567814.2, AD-568003.2, AD-569164.2, AD-569763.2, AD-565281.2, AD-571539.2, AD-572389.2, AD-567315.2, AD-571752.2, AD-568026.2, AD-571298, AD-572110.2, AD-572062.2, AD-572388.2, AD-572040.2, AD-567713.2, AD-567521.2, AD-567066.2, AD-1181519, AD-569268, or AD-570714.


In one embodiment, the antisense strand comprises at least 15 contiguous nucleotides differing by nor more than 0, 1, 2, or 3 nucleotides from any one of the antisense strand nucleotide sequences of a duplex selected from the group consisting of AD-1181519, AD-569268, or AD-570714. In another embodiment, the antisense strand comprises at least 15 contiguous nucleotides differing by nor more than 0, 1, 2, or 3 nucleotides from the antisense strand nucleotide sequences of a AD-570714.


In one embodiment, the dsRNA agent comprises at least one modified nucleotide.


In one embodiment, substantially all of the nucleotides of the sense strand; substantially all of the nucleotides of the antisense strand comprise a modification; or substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand comprise a modification.


In one embodiment, all of the nucleotides of the sense strand comprise a modification; all of the nucleotides of the antisense strand comprise a modification; or all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a modification.


In one embodiment, at least one of the modified nucleotides is selected from the group consisting of a deoxy-nucleotide, a 3′-terminal deoxy-thymine (dT) nucleotide, a 2′-O-methyl modified nucleotide, a 2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an unlocked nucleotide, a conformationally restricted nucleotide, a constrained ethyl nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-O-allyl-modified nucleotide, 2′-C-alkyl-modified nucleotide, 2′-hydroxly-modified nucleotide, a 2′-methoxyethyl modified nucleotide, a 2′-O-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, a non-natural base comprising nucleotide, a tetrahydropyran modified nucleotide, a 1,5-anhydrohexitol modified nucleotide, a cyclohexenyl modified nucleotide, a nucleotide comprising a phosphorothioate group, a nucleotide comprising a methylphosphonate group, a nucleotide comprising a 5′-phosphate, a nucleotide comprising a 5′-phosphate mimic, a thermally destabilizing nucleotide, a glycol modified nucleotide (GNA), and a 2-O—(N-methylacetamide) modified nucleotide; and combinations thereof.


In one embodiment, the modifications on the nucleotides are selected from the group consisting of LNA, HNA, CeNA, 2′-methoxyethyl, 2′-O-alkyl, 2′-O-allyl, 2′-C-allyl, 2′-fluoro, 2′-deoxy, 2′-hydroxyl, and glycol; and combinations thereof.


In one embodiment, at least one of the modified nucleotides is selected from the group consisting of a deoxy-nucleotide, a 2′-O-methyl modified nucleotide, a 2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a glycol modified nucleotide (GNA), e.g., Ggn, Cgn, Tgn, or Agn, and, a vinyl-phosphonate nucleotide; and combinations thereof.


In another embodiment, at least one of the modifications on the nucleotides is a thermally destabilizing nucleotide modification.


In one embodiment, the thermally destabilizing nucleotide modification is selected from the group consisting of an abasic modification; a mismatch with the opposing nucleotide in the duplex; and destabilizing sugar modification, a 2′-deoxy modification, an acyclic nucleotide, an unlocked nucleic acids (UNA), and a glycerol nucleic acid (GNA) The double stranded region may be 19-30 nucleotide pairs in length; 19-25 nucleotide pairs in length; 19-23 nucleotide pairs in length; 23-27 nucleotide pairs in length; or 21-23 nucleotide pairs in length.


In one embodiment, each strand is independently no more than 30 nucleotides in length.


In one embodiment, the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.


The region of complementarity may be at least 17 nucleotides in length; between 19 and 23 nucleotides in length; or 19 nucleotides in length.


In one embodiment, at least one strand comprises a 3′ overhang of at least 1 nucleotide. In another embodiment, at least one strand comprises a 3′ overhang of at least 2 nucleotides.


In one embodiment, the dsRNA agent further comprises a ligand.


In one embodiment, the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent.


In one embodiment, the ligand is an N-acetylgalactosamine (GalNAc) derivative.


In one embodiment, the ligand is one or more GalNAc derivatives attached through a monovalent, bivalent, or trivalent branched linker.


In one embodiment the ligand is




embedded image


In one embodiment, the dsRNA agent is conjugated to the ligand as shown in the following schematic




embedded image


and, wherein X is O or S.


In one embodiment, the X is O.


In one embodiment, the dsRNA agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.


In one embodiment, the phosphorothioate or methylphosphonate internucleotide linkage is at the 3′-terminus of one strand, e.g., the antisense strand or the sense strand.


In another embodiment, the phosphorothioate or methylphosphonate internucleotide linkage is at the 5′-terminus of one strand, e.g., the antisense strand or the sense strand.


In one embodiment, the phosphorothioate or methylphosphonate internucleotide linkage is at the both the 5′- and 3′-terminus of one strand. In one embodiment, the strand is the antisense strand.


In one embodiment, the base pair at the 1 position of the 5′-end of the antisense strand of the duplex is an AU base pair.


The present invention also provides cells containing any of the dsRNA agents of the invention and pharmaceutical compositions comprising any of the dsRNA agents of the invention.


The pharmaceutical composition of the invention may include dsRNA agent in an unbuffered solution, e.g., saline or water, or the pharmaceutical composition of the invention may include the dsRNA agent is in a buffer solution, e.g., a buffer solution comprising acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof; or phosphate buffered saline (PBS).


In one aspect, the present invention provides a method of inhibiting expression of a complement component C3 gene in a cell. The method includes contacting the cell with any of the dsRNAs of the invention or any of the pharmaceutical compositions of the invention, thereby inhibiting expression of the complement component C3 gene in the cell.


In one embodiment, the cell is within a subject, e.g., a human subject, e.g., a subject having a complement component C3-associated disorder, such as a complement component C3-associated disorder selected from the group consisting of cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy.


In one embodiment, contacting the cell with the dsRNA agent inhibits the expression of complement component C3 by at least 50%, 60%, 70%, 80%, 90%, or 95%.


In one embodiment, inhibiting expression of complement component C3 decreases complement component C3 protein level in serum of the subject by at least 50%, 60%, 70%, 80%, 90%, or 95%.


In one aspect, the present invention provides a method of treating a subject having a disorder that would benefit from reduction in complement component C3 expression. The method includes administering to the subject a therapeutically effective amount of any of the dsRNAs of the invention or any of the pharmaceutical compositions of the invention, thereby treating the subject having the disorder that would benefit from reduction in complement component C3 expression.


In another aspect, the present invention provides a method of preventing at least one symptom in a subject having a disorder that would benefit from reduction in complement component C3 expression. The method includes administering to the subject a prophylactically effective amount of any of the dsRNAs of the invention or any of the pharmaceutical compositions of the invention, thereby preventing at least one symptom in the subject having the disorder that would benefit from reduction in complement component C3 expression.


In one embodiment, the disorder is a complement component C3-associated disorder, e.g., a complement component C3-associated disorder is selected from the group consisting of cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy.


In one embodiment, the complement component C3-associated disorder is cold agglutinin disease (CAD).


In one embodiment, the subject is human.


In one embodiment, the administration of the agent to the subject causes a decrease in hemolysis and/or a decrease in C3 protein accumulation.


In one embodiment, the dsRNA agent is administered to the subject at a dose of about 0.01 mg/kg to about 50 mg/kg.


In one embodiment, the dsRNA agent is administered to the subject subcutaneously.


In one embodiment, the methods of the invention include further determining the level of complement component C3 in a sample(s) from the subject.


In one embodiment, the level of complement component C3 in the subject sample(s) is a complement component C3 protein level in a blood or serum sample(s).


In one embodiment, the methods of the invention further include administering to the subject an additional therapeutic agent for treatment of hemolysis.


The present invention also provides kits comprising any of the dsRNAs of the invention or any of the pharmaceutical compositions of the invention, and optionally, instructions for use.





BRIEF DESCRIPTION OF THE DRAWINGS


FIG. 1 schematically depicts the three complement pathways: alternative, classical and lectin.



FIG. 2 is a graph showing C3 mRNA levels in mice (n=3 per group) subcutaneously administered a single 2 mg/kg dose of the indicated dsRNA duplexes, on day 14 post-dose. C3 mRNA levels are shown relative to control levels detected with PBS treatment.



FIG. 3 is a graph showing C3 mRNA levels in mice (n=3 per group) subcutaneously administered a single 2 mg/kg dose of the indicated dsRNA duplexes, on day 14 post-dose. C3 mRNA levels are shown relative to control levels detected with PBS treatment.



FIG. 4 is a graph showing C3 mRNA levels in mice (n=3 per group) subcutaneously administered a single 2 mg/kg dose of the indicated dsRNA duplexes, on day 14 post-dose. C3 mRNA levels are shown relative to control levels detected with PBS treatment.



FIG. 5 is a Table depicting the treatment groups of Cynomolgus monkeys subcutaneously administered a single 3 mg/kg or 25 mg/kg dose of the indicated dsRNA duplexes.



FIG. 6 is a graph showing the effect of subcutaneous administration of a single 3 mg/kg or 25 mg/kg dose of the indicated dsRNA duplexes on % C3 protein levels remaining normalized to average predose C3 protein level in the serum of Cynmologous. The baseline was adjusted to day 1 dosing for all groups.



FIG. 7 is a Table depicting the treatment groups of Cynomolgus monkeys subcutaneously administered a single 3 mg/kg dose of the indicated dsRNA duplexes.



FIG. 8 is a graph showing the effect of subcutaneous administration of a single 3 mg/kg or 25 mg/kg dose of the indicated dsRNA duplexes on % C3 protein levels remaining normalized to average predose C3 protein level in the serum of Cynmologous.



FIG. 9 is a Table depicting the treatment groups and timing of administration and biopsy of Cynomolgus monkeys subcutaneously administered a single 3 mg/kg, 9 mg/kg, or 25 mg/kg dose, or a multi-dose of 3 mg/kg (3×3) of the indicated dsRNA duplexes.



FIG. 10 is a graph showing the effect of subcutaneous administration of a single 3 mg/kg or 25 mg/kg dose of the indicated dsRNA duplexes on % C3 protein levels remaining normalized to average predose C3 protein level in the serum of Cynmologous. For Group 2, Day −6 on the graph corresponds to Day −27 and Day 1 is the day on which the duplex was administered.





DETAILED DESCRIPTION OF THE INVENTION

The present invention provides iRNA compositions which effect the RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of a complement component C3 gene. The gene may be within a cell, e.g., a cell within a subject, such as a human. The use of these iRNAs enables the targeted degradation of mRNAs of the corresponding gene (complement component C3 gene) in mammals.


The iRNAs of the invention have been designed to target the human complement component C3 gene, including portions of the gene that are conserved in the complement component C3 orthologs of other mammalian species. Without intending to be limited by theory, it is believed that a combination or sub-combination of the foregoing properties and the specific target sites or the specific modifications in these iRNAs confer to the iRNAs of the invention improved efficacy, stability, potency, durability, and safety.


Accordingly, the present invention provides methods for treating and preventing a complement component C3-associated disorder, e.g., cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy, using iRNA compositions which effect the RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of a complement component C3 gene.


The iRNAs of the invention include an RNA strand (the antisense strand) having a region which is up to about 30 nucleotides or less in length, e.g., 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length, which region is substantially complementary to at least part of an mRNA transcript of a complement component C3 gene.


In certain embodiments, one or both of the strands of the double stranded RNAi agents of the invention is up to 66 nucleotides in length, e.g., 36-66, 26-36, 25-36, 31-60, 22-43, 27-53 nucleotides in length, with a region of at least 19 contiguous nucleotides that is substantially complementary to at least a part of an mRNA transcript of a complement component C3 gene. In some embodiments, such iRNA agents having longer length antisense strands preferably may include a second RNA strand (the sense strand) of 20-60 nucleotides in length wherein the sense and antisense strands form a duplex of 18-30 contiguous nucleotides.


The use of iRNAs of the invention enables the targeted degradation of mRNAs of the corresponding gene (complement component C3 gene) in mammals. Using in vitro and in vivo assays, the present inventors have demonstrated that iRNAs targeting a C3 gene can potently mediate RNAi, resulting in significant inhibition of expression of a C3 gene. Thus, methods and compositions including these iRNAs are useful for treating a subject having a complement component C3-associated disorder, e.g., cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy.


Accordingly, the present invention provides methods and combination therapies for treating a subject having a disorder that would benefit from inhibiting or reducing the expression of a C3 gene, e.g., a complement component C3-associated disease, such as cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy, using iRNA compositions which effect the RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of a C3 gene.


The present invention also provides methods for preventing at least one symptom in a subject having a disorder that would benefit from inhibiting or reducing the expression of a C3 gene, e.g., cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy.


For example, in a subject having cold agglutinin disease (CAD), the methods of the present invention may prevent at least one symptom in the subject including, e.g., hemolysis, MAC deposition and tissue damage, inflammation (e.g., chronic inflammation); in a subject having warm autoimmune hemolytic anemia, the methods of the present invention may prevent at least one symptom in the subject including, e.g., hemolysis, inflammation (e.g., chronic inflammation), and MAC tissue damage; in a subject having paroxysmal nocturnal hemoglobinuria (PNH), the methods of the present invention may prevent at least one symptom in the subject including, e.g., hemolysis, inflammation (e.g., chronic inflammation), thrombosis, and deficient hematopoiesis; in a subject having lupis nephritis (LN), the methods of the present invention may prevent at least one symptom in the subject including, e.g., inflammation (e.g., chronic inflammation), hematuria, proteinuria, edema, hypertension, and renal failure; in a subject having bullous pemphigoid, the methods of the present invention may prevent at least one symptom in the subject including, e.g., blister formation, inflammation (e.g., chronic inflammation), C3 deposition, and MAC tissue damage; in a subject having pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), the methods of the present invention may prevent at least one symptom in the subject including, e.g., blister formation, inflammation (e.g., chronic inflammation), C3 deposition, and MAC tissue damage; and in a subject having C3 glomerulopathy, the methods of the present invention may prevent at least one symptom in the subject including, e.g., inflammation (e.g., chronic inflammation), hematuria, proteinuria, edema, hypertension, and renal failure.


The following detailed description discloses how to make and use compositions containing iRNAs to inhibit the expression of a complement component C3 gene as well as compositions, uses, and methods for treating subjects that would benefit from inhibition and/or reduction of the expression of a complement component C3 gene, e.g., subjects susceptible to or diagnosed with a complement component C3-associated disorder.


I. Definitions

In order that the present invention may be more readily understood, certain terms are first defined. In addition, it should be noted that whenever a value or range of values of a parameter are recited, it is intended that values and ranges intermediate to the recited values are also intended to be part of this invention.


The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element, e.g., a plurality of elements.


The term “including” is used herein to mean, and is used interchangeably with, the phrase “including but not limited to”.


The term “or” is used herein to mean, and is used interchangeably with, the term “and/or,” unless context clearly indicates otherwise. For example, “sense strand or antisense strand” is understood as “sense strand or antisense strand or sense strand and antisense strand.”


The term “about” is used herein to mean within the typical ranges of tolerances in the art. For example, “about” can be understood as about 2 standard deviations from the mean. In certain embodiments, about means±10%. In certain embodiments, about means±5%. When about is present before a series of numbers or a range, it is understood that “about” can modify each of the numbers in the series or range.


The term “at least” prior to a number or series of numbers is understood to include the number adjacent to the term “at least”, and all subsequent numbers or integers that could logically be included, as clear from context. For example, the number of nucleotides in a nucleic acid molecule must be an integer. For example, “at least 19 nucleotides of a 21 nucleotide nucleic acid molecule” means that 19, 20, or 21 nucleotides have the indicated property. When at least is present before a series of numbers or a range, it is understood that “at least” can modify each of the numbers in the series or range.


As used herein, “no more than” or “less than” is understood as the value adjacent to the phrase and logical lower values or integers, as logical from context, to zero. For example, a duplex with an overhang of “no more than 2 nucleotides” has a 2, 1, or 0 nucleotide overhang. When “no more than” is present before a series of numbers or a range, it is understood that “no more than” can modify each of the numbers in the series or range. As used herein, ranges include both the upper and lower limit.


In the event of a conflict between a sequence and its indicated site on a transcript or other sequence, the nucleotide sequence recited in the specification takes precedence.


As used herein, the term “Complement Component 3,” used interchangeably with the term “C3,” refers to the well-known gene and polypeptide, also known in the art as ARMD9, C3a Anaphylatoxin, ASP, Complement Component C3a, C3a, Complement Component C3b, C3b, prepro-C3, Acylation-Stimulating Protein Cleavage Product, CPAMD1, Complement C3, C3 And PZP-Like Alpha-2-Macroglobulin Domain-Containing Protein 1, Complement Component C3, and AHUS5.


The term “C3” includes human C3, the amino acid and nucleotide sequence of which may be found in, for example, GenBank Accession No. NM_000064.3 (GI:726965399; SEQ ID NO:1); mouse C3, the amino acid and nucleotide sequence of which may be found in, for example, GenBank Accession No. NM_009778.3 (GI:773669943; SEQ ID NO:2); and rat C3, the amino acid and nucleotide sequence of which may be found in, for example, GenBank Accession No. NM_016994.2 (GI:158138560; SEQ ID NO:3).


The term “C3” also includes Macaca fascicularis C3, the amino acid and nucleotide sequence of which may be found in, for example, GenBank Accession No. XM_005587719.2 (GI:982312947; SEQ ID NO:4) and in the entry for the gene, ENSP00000245907 (locus=chr19:6921416:6963034), in the Macaca genome project web site (http://macaque.genomics.org.cn/page/species/index.jsp).


Additional examples of C3 mRNA sequences are readily available using, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site.


Exemplary C3 nucleotide sequences may also be found in SEQ ID NOs:1-8. SEQ ID NOs:5-8 are the reverse complement sequences of SEQ ID NOs:1-4, respectively.


Further information on C3 is provided, for example in the NCBI Gene database at http://www.ncbi.nlm.nih.gov/gene/718.


The entire contents of each of the foregoing GenBank Accession numbers and the Gene database numbers are incorporated herein by reference as of the date of filing this application.


The terms “complement component C3” and “C3,” as used herein, also refers to naturally occurring DNA sequence variations of the C3 gene. Numerous seuqnce variations within the C3 gene have been identified and may be found at, for example, NCBI dbSNP and UniProt (see, e.g., http://www.ncbi.nlm.nih.gov/snp?LinkName=gene_snp&from_uid=718, the entire contents of which is incorporated herein by reference as of the date of filing this application.


As used herein, “target sequence” refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a complement component C3 gene, including mRNA that is a product of RNA processing of a primary transcription product. The target portion of the sequence will be at least long enough to serve as a substrate for iRNA-directed cleavage at or near that portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a complement component C3 gene. In one embodiment, the target sequence is within the protein coding region of complement component C3.


The target sequence may be from about 19-36 nucleotides in length, e.g., preferably about 19-30 nucleotides in length. For example, the target sequence can be about 19-30 nucleotides, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the invention.


As used herein, the term “strand comprising a sequence” refers to an oligonucleotide comprising a chain of nucleotides that is described by the sequence referred to using the standard nucleotide nomenclature.


“G,” “C,” “A,” “T,” and “U” each generally stand for a nucleotide that contains guanine, cytosine, adenine, thymidine, and uracil as a base, respectively. However, it will be understood that the term “ribonucleotide” or “nucleotide” can also refer to a modified nucleotide, as further detailed below, or a surrogate replacement moiety (see, e.g., Table 1). The skilled person is well aware that guanine, cytosine, adenine, and uracil can be replaced by other moieties without substantially altering the base pairing properties of an oligonucleotide comprising a nucleotide bearing such replacement moiety. For example, without limitation, a nucleotide comprising inosine as its base can base pair with nucleotides containing adenine, cytosine, or uracil. Hence, nucleotides containing uracil, guanine, or adenine can be replaced in the nucleotide sequences of dsRNA featured in the invention by a nucleotide containing, for example, inosine. In another example, adenine and cytosine anywhere in the oligonucleotide can be replaced with guanine and uracil, respectively to form G-U Wobble base pairing with the target mRNA. Sequences containing such replacement moieties are suitable for the compositions and methods featured in the invention.


The terms “iRNA”, “RNAi agent,” “iRNA agent,”, “RNA interference agent” as used interchangeably herein, refer to an agent that contains RNA as that term is defined herein, and which mediates the targeted cleavage of an RNA transcript via an RNA-induced silencing complex (RISC) pathway. iRNA directs the sequence-specific degradation of mRNA through a process known as RNA interference (RNAi). The iRNA modulates, e.g., inhibits, the expression of a complement component C3 gene in a cell, e.g., a cell within a subject, such as a mammalian subject.


In one embodiment, an RNAi agent of the invention includes a single stranded RNA that interacts with a target RNA sequence, e.g., a complement component C3 target mRNA sequence, to direct the cleavage of the target RNA. Without wishing to be bound by theory it is believed that long double stranded RNA introduced into cells is broken down into siRNA by a Type III endonuclease known as Dicer (Sharp et al. (2001) Genes Dev. 15:485). Dicer, a ribonuclease-III-like enzyme, processes the dsRNA into 19-23 base pair short interfering RNAs with characteristic two base 3′ overhangs (Bernstein, et al., (2001) Nature 409:363). The siRNAs are then incorporated into an RNA-induced silencing complex (RISC) where one or more helicases unwind the siRNA duplex, enabling the complementary antisense strand to guide target recognition (Nykanen, et al., (2001) Cell 107:309). Upon binding to the appropriate target mRNA, one or more endonucleases within the RISC cleave the target to induce silencing (Elbashir, et al., (2001) Genes Dev. 15:188). Thus, in one aspect the invention relates to a single stranded RNA (siRNA) generated within a cell and which promotes the formation of a RISC complex to effect silencing of the target gene, i.e., a complement component C3 gene. Accordingly, the term “siRNA” is also used herein to refer to an iRNA as described above.


In certain embodiments, the RNAi agent may be a single-stranded siRNA (ssRNAi) that is introduced into a cell or organism to inhibit a target mRNA. Single-stranded RNAi agents bind to the RISC endonuclease, Argonaute 2, which then cleaves the target mRNA. The single-stranded siRNAs are generally 15-30 nucleotides and are chemically modified. The design and testing of single-stranded siRNAs are described in U.S. Pat. No. 8,101,348 and in Lima et al., (2012) Cell 150:883-894, the entire contents of each of which are hereby incorporated herein by reference. Any of the antisense nucleotide sequences described herein may be used as a single-stranded siRNA as described herein or as chemically modified by the methods described in Lima et al., (2012) Cell 150:883-894.


In certain embodiments, an “iRNA” for use in the compositions, uses, and methods of the invention is a double stranded RNA and is referred to herein as a “double stranded RNA agent,” “double stranded RNA (dsRNA) molecule,” “dsRNA agent,” or “dsRNA”. The term “dsRNA”, refers to a complex of ribonucleic acid molecules, having a duplex structure comprising two anti-parallel and substantially complementary nucleic acid strands, referred to as having “sense” and “antisense” orientations with respect to a target RNA, i.e., a complement component C3 gene. In some embodiments of the invention, a double stranded RNA (dsRNA) triggers the degradation of a target RNA, e.g., an mRNA, through a post-transcriptional gene-silencing mechanism referred to herein as RNA interference or RNAi.


In general, the majority of nucleotides of each strand of a dsRNA molecule are ribonucleotides, but as described in detail herein, each or both strands can also include one or more non-ribonucleotides, e.g., a deoxyribonucleotide or a modified nucleotide. In addition, as used in this specification, an “iRNA” may include ribonucleotides with chemical modifications; an iRNA may include substantial modifications at multiple nucleotides. As used herein, the term “modified nucleotide” refers to a nucleotide having, independently, a modified sugar moiety, a modified internucleotide linkage, or modified nucleobase, or any combination thereof. Thus, the term modified nucleotide encompasses substitutions, additions or removal of, e.g., a functional group or atom, to internucleoside linkages, sugar moieties, or nucleobases. The modifications suitable for use in the agents of the invention include all types of modifications disclosed herein or known in the art. Any such modifications, as used in a siRNA type molecule, are encompassed by “iRNA” or “RNAi agent” for the purposes of this specification and claims.


The duplex region may be of any length that permits specific degradation of a desired target RNA through a RISC pathway, and may range from about 19 to 36 base pairs in length, e.g., about 19-30 base pairs in length, for example, about 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, or 36 base pairs in length, such as about 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs in length. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the invention.


The two strands forming the duplex structure may be different portions of one larger RNA molecule, or they may be separate RNA molecules. Where the two strands are part of one larger molecule, and therefore are connected by an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′-end of the respective other strand forming the duplex structure, the connecting RNA chain is referred to as a “hairpin loop.” A hairpin loop can comprise at least one unpaired nucleotide. In some embodiments, the hairpin loop can comprise at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 23 or more unpaired nucleotides. In some embodiments, the hairpin loop can be 10 or fewer nucleotides. In some embodiments, the hairpin loop can be 8 or fewer unpaired nucleotides. In some embodiments, the hairpin loop can be 4-10 unpaired nucleotides. In some embodiments, the hairpin loop can be 4-8 nucleotides.


Where the two substantially complementary strands of a dsRNA are comprised by separate RNA molecules, those molecules need not be, but can be covalently connected. Where the two strands are connected covalently by means other than an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′-end of the respective other strand forming the duplex structure, the connecting structure is referred to as a “linker.” The RNA strands may have the same or a different number of nucleotides. The maximum number of base pairs is the number of nucleotides in the shortest strand of the dsRNA minus any overhangs that are present in the duplex. In addition to the duplex structure, an RNAi may comprise one or more nucleotide overhangs.


In certain embodiments, an iRNA agent of the invention is a dsRNA, each strand of which comprises 19-23 nucleotides, that interacts with a target RNA sequence, e.g., a complement component C3 gene, to direct cleavage of the target RNA.


In some embodiments, an iRNA of the invention is a dsRNA of 24-30 nucleotides that interacts with a target RNA sequence, e.g., a complement component C3 target mRNA sequence, to direct the cleavage of the target RNA.


As used herein, the term “nucleotide overhang” refers to at least one unpaired nucleotide that protrudes from the duplex structure of a double stranded iRNA. For example, when a 3-end of one strand of a dsRNA extends beyond the 5-end of the other strand, or vice versa, there is a nucleotide overhang. A dsRNA can comprise an overhang of at least one nucleotide; alternatively the overhang can comprise at least two nucleotides, at least three nucleotides, at least four nucleotides, at least five nucleotides or more. A nucleotide overhang can comprise or consist of a nucleotide/nucleoside analog, including a deoxynucleotide/nucleoside. The overhang(s) can be on the sense strand, the antisense strand, or any combination thereof. Furthermore, the nucleotide(s) of an overhang can be present on the 5′-end, 3′-end, or both ends of either an antisense or sense strand of a dsRNA.


In certain embodiments, the antisense strand of a dsRNA has a 1-10 nucleotides, e.g., a 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotide, overhang at the 3′-end or the 5′-end. In certain embodiments, the overhang on the sense strand or the antisense strand, or both, can include extended lengths longer than 10 nucleotides, e.g., 1-30 nucleotides, 2-30 nucleotides, 10-30 nucleotides, 10-25 nucleotides, 10-20 nucleotides, or 10-15 nucleotides in length. In certain embodiments, an extended overhang is on the sense strand of the duplex. In certain embodiments, an extended overhang is present on the 3′ end of the sense strand of the duplex. In certain embodiments, an extended overhang is present on the 5′ end of the sense strand of the duplex. In certain embodiments, an extended overhang is on the antisense strand of the duplex. In certain embodiments, an extended overhang is present on the 3′ end of the antisense strand of the duplex. In certain embodiments, an extended overhang is present on the 5′ end of the antisense strand of the duplex. In certain embodiments, one or more of the nucleotides in the extended overhang is replaced with a nucleoside thiophosphate. In certain embodiments, the overhang includes a self-complementary portion such that the overhang is capable of forming a hairpin structure that is stable under physiological conditions.


“Blunt” or “blunt end” means that there are no unpaired nucleotides at that end of the double stranded RNA agent, i.e., no nucleotide overhang. A “blunt ended” double stranded RNA agent is double stranded over its entire length, i.e., no nucleotide overhang at either end of the molecule. The RNAi agents of the invention include RNAi agents with no nucleotide overhang at one end (i.e., agents with one overhang and one blunt end) or with no nucleotide overhangs at either end. Most often such a molecule will be double-stranded over its entire length.


The term “antisense strand” or “guide strand” refers to the strand of an iRNA, e.g., a dsRNA, which includes a region that is substantially complementary to a target sequence, e.g., a complement component C3 mRNA.


As used herein, the term “region of complementarity” refers to the region on the antisense strand that is substantially complementary to a sequence, for example a target sequence, e.g., a complement component C3 nucleotide sequence, as defined herein. Where the region of complementarity is not fully complementary to the target sequence, the mismatches can be in the internal or terminal regions of the molecule. Generally, the most tolerated mismatches are in the terminal regions, e.g., within 5, 4, or 3 nucleotides of the 5′- or 3′-end of the iRNA. In some embodiments, a double stranded RNA agent of the invention includes a nucleotide mismatch in the antisense strand. In some embodiments, the antisense strand of the double stranded RNA agent of the invention includes no more than 4 mismatches with the target mRNA, e.g., the antisense strand includes 4, 3, 2, 1, or 0 mismatches with the target mRNA. In some embodiments, the antisense strand double stranded RNA agent of the invention includes no more than 4 mismatches with the sense strand, e.g., the antisense strand includes 4, 3, 2, 1, or 0 mismatches with the sense strand. In some embodiments, a double stranded RNA agent of the invention includes a nucleotide mismatch in the sense strand. In some embodiments, the sense strand of the double stranded RNA agent of the invention includes no more than 4 mismatches with the antisense strand, e.g., the sense strand includes 4, 3, 2, 1, or 0 mismatches with the antisense strand. In some embodiments, the nucleotide mismatch is, for example, within 5, 4, 3 nucleotides from the 3′-end of the iRNA. In another embodiment, the nucleotide mismatch is, for example, in the 3′-terminal nucleotide of the iRNA agent. In some embodiments, the mismatch(s) is not in the seed region.


Thus, an RNAi agent as described herein can contain one or more mismatches to the target sequence. In one embodiment, a RNAi agent as described herein contains no more than 3 mismatches (i.e., 3, 2, 1, or 0 mismatches). In one embodiment, an RNAi agent as described herein contains no more than 2 mismatches. In one embodiment, an RNAi agent as described herein contains no more than 1 mismatch. In one embodiment, an RNAi agent as described herein contains 0 mismatches. In certain embodiments, if the antisense strand of the RNAi agent contains mismatches to the target sequence, the mismatch can optionally be restricted to be within the last 5 nucleotides from either the 5′- or 3′-end of the region of complementarity. For example, in such embodiments, for a 23 nucleotide RNAi agent, the strand which is complementary to a region of a C3 gene, generally does not contain any mismatch within the central 13 nucleotides. The methods described herein or methods known in the art can be used to determine whether an RNAi agent containing a mismatch to a target sequence is effective in inhibiting the expression of a C3 gene. Consideration of the efficacy of RNAi agents with mismatches in inhibiting expression of a C3 gene is important, especially if the particular region of complementarity in a C3 gene is known to have polymorphic sequence variation within the population.


The term “sense strand” or “passenger strand” as used herein, refers to the strand of an iRNA that includes a region that is substantially complementary to a region of the antisense strand as that term is defined herein.


As used herein, “substantially all of the nucleotides are modified” are largely but not wholly modified and can include not more than 5, 4, 3, 2, or 1 unmodified nucleotides.


As used herein, the term “cleavage region” refers to a region that is located immediately adjacent to the cleavage site. The cleavage site is the site on the target at which cleavage occurs. In some embodiments, the cleavage region comprises three bases on either end of, and immediately adjacent to, the cleavage site. In some embodiments, the cleavage region comprises two bases on either end of, and immediately adjacent to, the cleavage site. In some embodiments, the cleavage site specifically occurs at the site bound by nucleotides 10 and 11 of the antisense strand, and the cleavage region comprises nucleotides 11, 12 and 13.


As used herein, and unless otherwise indicated, the term “complementary,” when used to describe a first nucleotide sequence in relation to a second nucleotide sequence, refers to the ability of an oligonucleotide or polynucleotide comprising the first nucleotide sequence to hybridize and form a duplex structure under certain conditions with an oligonucleotide or polynucleotide comprising the second nucleotide sequence, as will be understood by the skilled person. Such conditions can, for example, be stringent conditions, where stringent conditions can include: 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA, 50° C. or 70° C. for 12-16 hours followed by washing (see, e.g., “Molecular Cloning: A Laboratory Manual, Sambrook, et al. (1989) Cold Spring Harbor Laboratory Press). Other conditions, such as physiologically relevant conditions as can be encountered inside an organism, can apply. The skilled person will be able to determine the set of conditions most appropriate for a test of complementarity of two sequences in accordance with the ultimate application of the hybridized nucleotides.


Complementary sequences within an iRNA, e.g., within a dsRNA as described herein, include base-pairing of the oligonucleotide or polynucleotide comprising a first nucleotide sequence to an oligonucleotide or polynucleotide comprising a second nucleotide sequence over the entire length of one or both nucleotide sequences. Such sequences can be referred to as “fully complementary” with respect to each other herein. However, where a first sequence is referred to as “substantially complementary” with respect to a second sequence herein, the two sequences can be fully complementary, or they can form one or more, but generally not more than 5, 4, 3, or 2 mismatched base pairs upon hybridization for a duplex up to 30 base pairs, while retaining the ability to hybridize under the conditions most relevant to their ultimate application, e.g., inhibition of gene expression via a RISC pathway. However, where two oligonucleotides are designed to form, upon hybridization, one or more single stranded overhangs, such overhangs shall not be regarded as mismatches with regard to the determination of complementarity. For example, a dsRNA comprising one oligonucleotide 21 nucleotides in length and another oligonucleotide 23 nucleotides in length, wherein the longer oligonucleotide comprises a sequence of 21 nucleotides that is fully complementary to the shorter oligonucleotide, can yet be referred to as “fully complementary” for the purposes described herein.


“Complementary” sequences, as used herein, can also include, or be formed entirely from, non-Watson-Crick base pairs or base pairs formed from non-natural and modified nucleotides, in so far as the above requirements with respect to their ability to hybridize are fulfilled. Such non-Watson-Crick base pairs include, but are not limited to, G:U Wobble or Hoogstein base pairing.


The terms “complementary,” “fully complementary” and “substantially complementary” herein can be used with respect to the base matching between the sense strand and the antisense strand of a dsRNA, or between the antisense strand of a double stranded RNA agent and a target sequence, as will be understood from the context of their use.


As used herein, a polynucleotide that is “substantially complementary to at least part of” a messenger RNA (mRNA) refers to a polynucleotide that is substantially complementary to a contiguous portion of the mRNA of interest (e.g., an mRNA encoding a complement component C3 gene). For example, a polynucleotide is complementary to at least a part of a complement component C3 mRNA if the sequence is substantially complementary to a non-interrupted portion of an mRNA encoding a complement component C3 gene.


Accordingly, in some embodiments, the antisense polynucleotides disclosed herein are fully complementary to the target complement component C3 sequence. In other embodiments, the antisense polynucleotides disclosed herein are substantially complementary to the target complement component C3 sequence and comprise a contiguous nucleotide sequence which is at least 80% complementary over its entire length to the equivalent region of the nucleotide sequence of any one of SEQ ID NOs:1-4, or a fragment of any one of SEQ ID NOs:1-4, such as about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary.


In some embodiments, the antisense polynucleotides disclosed herein are substantially complementary to a fragment of a target complement component C3 sequence and comprise a contiguous nucleotide sequence which is at least 80% complementary over its entire length to a fragment of SEQ ID NO: 1 selected from the group of nucleotides 475-497, 487-509, 490-512, 491-513, 705-727, 809-831, 813-835, 1147-1169, 1437-1459, 1439-1461, 1447-1469, 2596-2618, 2634-2656, 3012-3034, 3334-3356, 3611-3633, 3614-3636, 3622-3655, 3809-3831, 3846-3868, 3847-3869, 3920-3942, 4047-4069, 4061-4083, 4156-4178, 4157-4177, 4162-4184, 4178-4200, 4226-4248, 4369-4391, 4392-4414, 4521-4543, 4522-4544, 4523-4545, 5012-5034 of SEQ ID NO: 1, such as about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary.


In other embodiments, the antisense polynucleotides disclosed herein are substantially complementary to a fragment of a target complement component C3 sequence and comprise a contiguous nucleotide sequence which is at least 80% complementary over its entire length to a fragment of SEQ ID NO: 1 selected from the group of nucleotides 705-727, 809-831, or 2634-2656 of SEQ ID NO: 1, such as about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary. In one embodiment, the antisense polynucleotides disclosed herein are substantially complementary to a fragment of a target complement component C3 sequence and comprise a contiguous nucleotide sequence which is at least 80% complementary over its entire length to a fragment of SEQ ID NO: 1 from nucleotides 2634-2656, such as about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary.


In other embodiments, the antisense polynucleotides disclosed herein are substantially complementary to the target C3 sequence and comprise a contiguous nucleotide sequence which is at least about 80% complementary over its entire length to any one of the sense strand nucleotide sequences in any one of any one of Tables 2-7, 15, 18, 20-23, 30, and 31, or a fragment of any one of the sense strand nucleotide sequences in any one of Tables 2-7, 15, 18, 20-23, 30, and 31, such as about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% complementary.


In one embodiment, an RNAi agent of the disclosure includes a sense strand that is substantially complementary to an antisense polynucleotide which, in turn, is the same as a target C3 sequence, and wherein the sense strand polynucleotide comprises a contiguous nucleotide sequence which is at least about 80% complementary over its entire length to the equivalent region of the nucleotide sequence of SEQ ID NOs: 5-8, or a fragment of any one of SEQ ID NOs:5-8, such as about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% complementary.


In some embodiments, an iRNA of the invention includes a sense strand that is substantially complementary to an antisense polynucleotide which, in turn, is complementary to a target complement component C3 sequence, and wherein the sense strand polynucleotide comprises a contiguous nucleotide sequence which is at least about 80% complementary over its entire length to any one of the antisense strand nucleotide sequences in any one of any one of Tables 2-7, 15, 18, 20-23, 30, and 31, or a fragment of any one of the antisense strand nucleotide sequences in any one of Tables 2-7, 15, 18, 20-23, 30, and 31, such as about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% complementary In certain embodiments, the sense and antisense strands are selected from any one of duplexes AD-565541.2, AD-564742, AD-567304, AD-568978, AD-569164, AD-569272.2, AD-569765.2, AD-564730.2, AD-567315, AD-564745.2, AD-571715.2, AD-570714, AD-571826, AD-572041.2, AD-572039.2, AD-572387, AD-568586.2, AD-566837.2, AD-566444.2, AD-567700.2, AD-567814.2, AD-568003.2, AD-569164.2, AD-569763.2, AD-565281.2, AD-571539.2, AD-572389.2, AD-567315.2, AD-571752.2, AD-568026.2, AD-571298, AD-572110.2, AD-572062.2, AD-572388.2, AD-572040.2, AD-567713.2, AD-567521.2, AD-567066.2, AD-1181519, AD-569268, or AD-570714.


In some embodiments, the sense and antisense strands are selected from any one of duplexes AD-1181519, AD-569268, or AD-570714. In one embodiment, the dulex is AD-570714.


In general, an “iRNA” includes ribonucleotides with chemical modifications. Such modifications may include all types of modifications disclosed herein or known in the art. Any such modifications, as used in a dsRNA molecule, are encompassed by “iRNA” for the purposes of this specification and claims.


In an aspect of the invention, an agent for use in the methods and compositions of the invention is a single-stranded antisense oligonucleotide molecule that inhibits a target mRNA via an antisense inhibition mechanism. The single-stranded antisense oligonucleotide molecule is complementary to a sequence within the target mRNA. The single-stranded antisense oligonucleotides can inhibit translation in a stoichiometric manner by base pairing to the mRNA and physically obstructing the translation machinery, see Dias, N. et al., (2002) Mol Cancer Ther 1:347-355. The single-stranded antisense oligonucleotide molecule may be about 14 to about 30 nucleotides in length and have a sequence that is complementary to a target sequence. For example, the single-stranded antisense oligonucleotide molecule may comprise a sequence that is at least about 14, 15, 16, 17, 18, 19, 20, or more contiguous nucleotides from any one of the antisense sequences described herein.


The phrase “contacting a cell with an iRNA,” such as a dsRNA, as used herein, includes contacting a cell by any possible means. Contacting a cell with an iRNA includes contacting a cell in vitro with the iRNA or contacting a cell in vivo with the iRNA. The contacting may be done directly or indirectly. Thus, for example, the iRNA may be put into physical contact with the cell by the individual performing the method, or alternatively, the iRNA may be put into a situation that will permit or cause it to subsequently come into contact with the cell.


Contacting a cell in vitro may be done, for example, by incubating the cell with the iRNA. Contacting a cell in vivo may be done, for example, by injecting the iRNA into or near the tissue where the cell is located, or by injecting the iRNA into another area, e.g., the bloodstream or the subcutaneous space, such that the agent will subsequently reach the tissue where the cell to be contacted is located. For example, the iRNA may contain or be coupled to a ligand, e.g., GalNAc, that directs the iRNA to a site of interest, e.g., the liver. Combinations of in vitro and in vivo methods of contacting are also possible. For example, a cell may also be contacted in vitro with an iRNA and subsequently transplanted into a subject.


In certain embodiments, contacting a cell with an iRNA includes “introducing” or “delivering the iRNA into the cell” by facilitating or effecting uptake or absorption into the cell. Absorption or uptake of an iRNA can occur through unaided diffusion or active cellular processes, or by auxiliary agents or devices. Introducing an iRNA into a cell may be in vitro or in vivo. For example, for in vivo introduction, iRNA can be injected into a tissue site or administered systemically. In vitro introduction into a cell includes methods known in the art such as electroporation and lipofection. Further approaches are described herein below or are known in the art.


The term “lipid nanoparticle” or “LNP” is a vesicle comprising a lipid layer encapsulating a pharmaceutically active molecule, such as a nucleic acid molecule, e.g., an iRNA or a plasmid from which an iRNA is transcribed. LNPs are described in, for example, U.S. Pat. Nos. 6,858,225, 6,815,432, 8,158,601, and 8,058,069, the entire contents of which are hereby incorporated herein by reference.


As used herein, a “subject” is an animal, such as a mammal, including a primate (such as a human, a non-human primate, e.g., a monkey, and a chimpanzee), a non-primate (such as a cow, a pig, a horse, a goat, a rabbit, a sheep, a hamster, a guinea pig, a cat, a dog, a rat, or a mouse), or a bird that expresses the target gene, either endogenously or heterologously. In an embodiment, the subject is a human, such as a human being treated or assessed for a disease or disorder that would benefit from reduction in complement component C3 expression; a human at risk for a disease or disorder that would benefit from reduction in C3 expression; a human having a disease or disorder that would benefit from reduction in complement component C3 expression; or human being treated for a disease or disorder that would benefit from reduction in complement component C3 expression as described herein. In some embodiments, the subject is a female human. In other embodiments, the subject is a male human. In one embodiment, the subject is an adult subject. In another embodiment, the subject is a pediatric subject.


As used herein, the terms “treating” or “treatment” refer to a beneficial or desired result, such as reducing at least one sign or symptom of a complement component C3-associated disorder, e.g., hemolysis in a subject. Treatment also includes a reduction of one or more sign or symptoms associated with unwanted complement component C3 expression, e.g., hemolysis; diminishing the extent of unwanted complement component C3 activation or stabilization; amelioration or palliation of unwanted complement component C3 activation or stabilization. “Treatment” can also mean prolonging survival as compared to expected survival in the absence of treatment.


The term “lower” in the context of the level of complement component C3 gene expression or complement component C3 protein production in a subject, or a disease marker or symptom refers to a statistically significant decrease in such level. The decrease can be, for example, at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or below the level of detection for the detection method in a relevant cell or tissue, e.g., a liver cell, or other subject sample, e.g., blood or serum derived therefrom, urine.


As used herein, “prevention” or “preventing,” when used in reference to a disease or disorder, that would benefit from a reduction in expression of a complement component C3 gene or production of complement component C3 protein, e.g., in a subject susceptible to a complement component C3-associated disorder due to, e.g., aging, genetic factors, hormone changes, diet, and a sedentary lifestyle. In certain embodiments, the disease or disorder is e.g., a symptom of unwanted C3 activation or stabilization, such as a hemolysis. The likelihood of developing, e.g., hemolysis, is reduced, for example, when an individual having one or more risk factors for hemolysis either fails to develop hemolysis or develops hemolysis with less severity relative to a population having the same risk factors and not receiving treatment as described herein. The failure to develop a complement component C3-associated disorder, e.g., hemolysis, or a delay in the time to develop hemolysis by months or years is considered effective prevention. Prevention may require administration of more than one dose if the iRNA agent.


As used herein, the term “complement component C3-associated disease” or “C3-associated disease,” is a disease or disorder that would benefit from reduction in complement component C3 expression. Non-limiting examples of complement component C3-associated diseases include, cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy.


A “therapeutically-effective amount” or “prophylactically effective amount” also includes an amount of an RNAi agent that produces some desired effect at a reasonable benefit/risk ratio applicable to any treatment. The iRNA employed in the methods of the present invention may be administered in a sufficient amount to produce a reasonable benefit/risk ratio applicable to such treatment.


The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human subjects and animal subjects without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.


The phrase “pharmaceutically-acceptable carrier” as used herein means a pharmaceutically-acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc magnesium, calcium or zinc stearate, or steric acid), or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject being treated. Such carriers are known in the art. Pharmaceutically acceptable carriers include carriers for administration by injection.


The term “sample,” as used herein, includes a collection of similar fluids, cells, or tissues isolated from a subject, as well as fluids, cells, or tissues present within a subject. Examples of biological fluids include blood, serum and serosal fluids, plasma, cerebrospinal fluid, ocular fluids, lymph, urine, saliva, and the like. Tissue samples may include samples from tissues, organs, or localized regions. For example, samples may be derived from particular organs, parts of organs, or fluids or cells within those organs. In certain embodiments, samples may be derived from the liver (e.g., whole liver or certain segments of liver or certain types of cells in the liver, such as, e.g., hepatocytes). In some embodiments, a “sample derived from a subject” refers to urine obtained from the subject. A “sample derived from a subject” can refer to blood or blood derived serum or plasma from the subject.


II. iRNAs of the Invention

The present invention provides iRNAs which inhibit the expression of a complement component C3 gene. In preferred embodiments, the iRNA includes double stranded ribonucleic acid (dsRNA) molecules for inhibiting the expression of a complement component C3 gene in a cell, such as a cell within a subject, e.g., a mammal, such as a human susceptible to developing a complement component C3-associated disorder, e.g., hemolysis. The dsRNAi agent includes an antisense strand having a region of complementarity which is complementary to at least a part of an mRNA formed in the expression of a complement component C3 gene. The region of complementarity is about 19-30 nucleotides in length (e.g., about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, or 19 nucleotides in length). Upon contact with a cell expressing the complement component C3 gene, the iRNA inhibits the expression of the complement component C3 gene (e.g., a human, a primate, a non-primate, or a rat complement component C3 gene) by at least about 50% as assayed by, for example, a PCR or branched DNA (bDNA)-based method, or by a protein-based method, such as by immunofluorescence analysis, using, for example, western blotting or flow cytometric techniques. In preferred embodiments, inhibition of expression is determined by the qPCR method provided in the examples, especially in Example 2 with the siRNA at a 10 nM concentration in an appropriate organism cell line provided therein. In preferred embodiments, inhibition of expression in vivo is determined by knockdown of the human gene in a rodent expressing the human gene, e.g., a mouse or an AAV-infected mouse expressing the human target gene, e.g., when administered as single dose, e.g., at 3 mg/kg at the nadir of RNA expression. RNA expression in liver is determined using the PCR methods provided in Example 2.


A dsRNA includes two RNA strands that are complementary and hybridize to form a duplex structure under conditions in which the dsRNA will be used. One strand of a dsRNA (the antisense strand) includes a region of complementarity that is substantially complementary, and generally fully complementary, to a target sequence. The target sequence can be derived from the sequence of an mRNA formed during the expression of a complement component C3 gene. The other strand (the sense strand) includes a region that is complementary to the antisense strand, such that the two strands hybridize and form a duplex structure when combined under suitable conditions. As described elsewhere herein and as known in the art, the complementary sequences of a dsRNA can also be contained as self-complementary regions of a single nucleic acid molecule, as opposed to being on separate oligonucleotides.


Generally, the duplex structure is 19 to 30 base pairs in length. Similarly, the region of complementarity to the target sequence is 19 to 30 nucleotides in length.


In some embodiments, the dsRNA is about 19 to about 23 nucleotides in length, or about 25 to about 30 nucleotides in length. In general, the dsRNA is long enough to serve as a substrate for the Dicer enzyme. For example, it is well-known in the art that dsRNAs longer than about 21-23 nucleotides in length may serve as substrates for Dicer. As the ordinarily skilled person will also recognize, the region of an RNA targeted for cleavage will most often be part of a larger RNA molecule, often an mRNA molecule. Where relevant, a “part” of an mRNA target is a contiguous sequence of an mRNA target of sufficient length to allow it to be a substrate for RNAi-directed cleavage (i.e., cleavage through a RISC pathway).


One of skill in the art will also recognize that the duplex region is a primary functional portion of a dsRNA, e.g., a duplex region of about 19 to about 30 base pairs, e.g., about 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs. Thus, in one embodiment, to the extent that it becomes processed to a functional duplex, of e.g., 15-30 base pairs, that targets a desired RNA for cleavage, an RNA molecule or complex of RNA molecules having a duplex region greater than 30 base pairs is a dsRNA. Thus, an ordinarily skilled artisan will recognize that in one embodiment, a miRNA is a dsRNA. In another embodiment, a dsRNA is not a naturally occurring miRNA. In another embodiment, an iRNA agent useful to target complement component C3 gene expression is not generated in the target cell by cleavage of a larger dsRNA.


A dsRNA as described herein can further include one or more single-stranded nucleotide overhangs e.g., 1-4, 2-4, 1-3, 2-3, 1, 2, 3, or 4 nucleotides. dsRNAs having at least one nucleotide overhang can have superior inhibitory properties relative to their blunt-ended counterparts. A nucleotide overhang can comprise or consist of a nucleotide/nucleoside analog, including a deoxynucleotide/nucleoside. The overhang(s) can be on the sense strand, the antisense strand, or any combination thereof. Furthermore, the nucleotide(s) of an overhang can be present on the 5′-end, 3′-end, or both ends of an antisense or sense strand of a dsRNA.


A dsRNA can be synthesized by standard methods known in the art. Double stranded RNAi compounds of the invention may be prepared using a two-step procedure. First, the individual strands of the double stranded RNA molecule are prepared separately. Then, the component strands are annealed. The individual strands of the siRNA compound can be prepared using solution-phase or solid-phase organic synthesis or both. Organic synthesis offers the advantage that the oligonucleotide strands comprising unnatural or modified nucleotides can be easily prepared. Similarly, single-stranded oligonucleotides of the invention can be prepared using solution-phase or solid-phase organic synthesis or both.


In an aspect, a dsRNA of the invention includes at least two nucleotide sequences, a sense sequence and an anti-sense sequence. The sense strand is selected from the group of sequences provided in any one of Tables 2-7, 15, 18, 20-23, 30, or 31, and the corresponding antisense strand of the sense strand is selected from the group of sequences of any one of Tables 2-7, 15, 18, 20-23, 30, or 31. In this aspect, one of the two sequences is complementary to the other of the two sequences, with one of the sequences being substantially complementary to a sequence of an mRNA generated in the expression of a complement component C3 gene. As such, in this aspect, a dsRNA will include two oligonucleotides, where one oligonucleotide is described as the sense strand in any one of Tables 2-7, 15, 18, 20-23, 30, or 31, and the second oligonucleotide is described as the corresponding antisense strand of the sense strand in any one of Tables 2-7, 15, 18, 20-23, 30, or 31. In certain embodiments, the substantially complementary sequences of the dsRNA are contained on separate oligonucleotides. In other embodiments, the substantially complementary sequences of the dsRNA are contained on a single oligonucleotide. In certain embodiments, the sense or antisense strand is selected from the sense or antisense strand of any one of duplexes AD-565541.2, AD-564742, AD-567304, AD-568978, AD-569164, AD-569272.2, AD-569765.2, AD-564730.2, AD-567315, AD-564745.2, AD-571715.2, AD-570714, AD-571826, AD-572041.2, AD-572039.2, AD-572387, AD-568586.2, AD-566837.2, AD-566444.2, AD-567700.2, AD-567814.2, AD-568003.2, AD-569164.2, AD-569763.2, AD-565281.2, AD-571539.2, AD-572389.2, AD-567315.2, AD-571752.2, AD-568026.2, AD-571298, AD-572110.2, AD-572062.2, AD-572388.2, AD-572040.2, AD-567713.2, AD-567521.2, AD-567066.2, AD-1181519, AD-569268, or AD-570714. In other embodiment, the sense or antisense strand is selected from the sense or antisense strand of any one of duplexes AD-1181519, AD-569268, or AD-570714. In one embodiment, the duplex is AD-570714.


It will be understood that, although the sequences in Tables 2, 4, 6, 20, 22, and 30 are not described as modified or conjugated sequences, the RNA of the iRNA of the invention e.g., a dsRNA of the invention, may comprise any one of the sequences set forth in any one of Tables 3, 5, 7, 15, 18, 21, 23, or 31 that is un-modified, un-conjugated, or modified or conjugated differently than described therein. In other words, the invention encompasses dsRNA of Tables 2-7, 15, 18, 20-23, 30, or 31 which are un-modified, un-conjugated, modified, or conjugated, as described herein.


The skilled person is well aware that dsRNAs having a duplex structure of about 20 to 23 base pairs, e.g., 21, base pairs have been hailed as particularly effective in inducing RNA interference (Elbashir et al., EMBO 2001, 20:6877-6888). However, others have found that shorter or longer RNA duplex structures can also be effective (Chu and Rana (2007) RNA 14:1714-1719; Kim et al. (2005) Nat Biotech 23:222-226). In the embodiments described above, by virtue of the nature of the oligonucleotide sequences provided in any one of Tables 2-7, 15, 18, 20-23, 30, or 31, dsRNAs described herein can include at least one strand of a length of minimally 21 nucleotides. It can be reasonably expected that shorter duplexes having any one of the sequences in any one of Tables 2-7, 15, 18, 20-23, 30, or 31 minus only a few nucleotides on one or both ends can be similarly effective as compared to the dsRNAs described above. Hence, dsRNAs having a sequence of at least 19, 20, or more contiguous nucleotides derived from any one of the sequences of any one of Tables 2-7, 15, 18, 20-23, 30, or 31, and differing in their ability to inhibit the expression of a complement component C3 gene by not more than about 5, 10, 15, 20, 25, or 30% inhibition from a dsRNA comprising the full sequence, are contemplated to be within the scope of the present invention.


In addition, the RNAs provided in Tables 2-7, 15, 18, 20-23, 30, or 31 identify a site(s) in a complement component C3 transcript that is susceptible to RISC-mediated cleavage. As such, the present invention further features iRNAs that target within one of these sites. As used herein, an iRNA is said to target within a particular site of an RNA transcript if the iRNA promotes cleavage of the transcript anywhere within that particular site. Such an iRNA will generally include at least about 19 contiguous nucleotides from any one of the sequences provided in any one of Tables 2-7, 15, 18, 20-23, 30, or 31 coupled to additional nucleotide sequences taken from the region contiguous to the selected sequence in a complement component C3 gene.


III. Modified iRNAs of the Invention

In certain embodiments, the RNA of the iRNA of the invention e.g., a dsRNA, is un-modified, and does not comprise, e.g., chemical modifications or conjugations known in the art and described herein. In other embodiments, the RNA of an iRNA of the invention, e.g., a dsRNA, is chemically modified to enhance stability or other beneficial characteristics. In certain embodiments of the invention, substantially all of the nucleotides of an iRNA of the invention are modified. In other embodiments of the invention, all of the nucleotides of an iRNA or substantially all of the nucleotides of an iRNA are modified, i.e., not more than 5, 4, 3, 2, or lunmodified nucleotides are present in a strand of the iRNA.


The nucleic acids featured in the invention can be synthesized or modified by methods well established in the art, such as those described in “Current protocols in nucleic acid chemistry,” Beaucage, S. L. et al. (Edrs.), John Wiley & Sons, Inc., New York, NY, USA, which is hereby incorporated herein by reference. Modifications include, for example, end modifications, e.g., 5′-end modifications (phosphorylation, conjugation, inverted linkages) or 3′-end modifications (conjugation, DNA nucleotides, inverted linkages, etc.); base modifications, e.g., replacement with stabilizing bases, destabilizing bases, or bases that base pair with an expanded repertoire of partners, removal of bases (abasic nucleotides), or conjugated bases; sugar modifications (e.g., at the 2′-position or 4′-position) or replacement of the sugar; or backbone modifications, including modification or replacement of the phosphodiester linkages. Specific examples of iRNA compounds useful in the embodiments described herein include, but are not limited to RNAs containing modified backbones or no natural internucleoside linkages. RNAs having modified backbones include, among others, those that do not have a phosphorus atom in the backbone. For the purposes of this specification, and as sometimes referenced in the art, modified RNAs that do not have a phosphorus atom in their internucleoside backbone can also be considered to be oligonucleosides. In some embodiments, a modified iRNA will have a phosphorus atom in its internucleoside backbone.


Modified RNA backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates including 3-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates including 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′. Various salts, mixed salts and free acid forms are also included. In some embodiments of the invention, the dsRNA agents of the invention are in a free acid form. In other embodiments of the invention, the dsRNA agents of the invention are in a salt form. In one embodiment, the dsRNA agents of the invention are in a sodium salt form. In certain embodiments, when the dsRNA agents of the invention are in the sodium salt form, sodium ions are present in the agent as counterions for substantially all of the phosphodiester and/or phosphorothiotate groups present in the agent. Agents in which substantially all of the phosphodiester and/or phosphorothioate linkages have a sodium counterion include not more than 5, 4, 3, 2, or 1 phosphodiester and/or phosphorothioate linkages without a sodium counterion. In some embodiments, when the dsRNA agents of the invention are in the sodium salt form, sodium ions are present in the agent as counterions for all of the phosphodiester and/or phosphorothiotate groups present in the agent.


Representative U.S. patents that teach the preparation of the above phosphorus-containing linkages include, but are not limited to, U.S. Pat. Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243; 5,177,195; 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; 5,541,316; 5,550,111; 5,563,253; 5,571,799; 5,587,361; 5,625,050; 6,028,188; 6,124,445; 6,160,109; 6,169,170; 6,172,209; 6,239,265; 6,277,603; 6,326,199; 6,346,614; 6,444,423; 6,531,590; 6,534,639; 6,608,035; 6,683,167; 6,858,715; 6,867,294; 6,878,805; 7,015,315; 7,041,816; 7,273,933; 7,321,029; and U.S. Pat. RE39464, the entire contents of each of which are hereby incorporated herein by reference.


Modified RNA backbones that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatoms and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages. These include 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.


Representative U.S. patents that teach the preparation of the above oligonucleosides include, but are not limited to, U.S. Pat. Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033; 5,64,562; 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; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and 5,677,439, the entire contents of each of which are hereby incorporated herein by reference.


Suitable RNA mimetics are contemplated for use in iRNAs provided herein, in which both the sugar and the internucleoside linkage, i.e., 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 in which an RNA 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 RNA is replaced with an amide containing backbone, in particular 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 US 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, the entire contents of each of which are hereby incorporated herein by reference. Additional PNA compounds suitable for use in the iRNAs of the invention are described in, for example, in Nielsen et al., Science, 1991, 254, 1497-1500.


Some embodiments featured in the invention include RNAs with phosphorothioate backbones and oligonucleosides with heteroatom backbones, and in particular —CH2—NH—CH2—, —CH2—N(CH3)—O—CH2— [known as a methylene (methylimino) or MMI backbone], —CH2—O—N(CH3)—CH2—, —CH2—N(CH3)—N(CH3)—CH2— and —N(CH3)—CH2—CH2— [wherein the native phosphodiester backbone is represented as —O—P—O—CH2— ] of the above-referenced U.S. Pat. No. 5,489,677, and the amide backbones of the above-referenced U.S. Pat. No. 5,602,240. In some embodiments, the RNAs featured herein have morpholino backbone structures of the above-referenced U.S. Pat. No. 5,034,506.


Modified RNAs can also contain one or more substituted sugar moieties. The iRNAs, e.g., dsRNAs, featured herein can include one of the following at the 2′-position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynyl can be substituted or unsubstituted C1 to C10 alkyl or C2 to C10 alkenyl and alkynyl. Exemplary suitable modifications include O[(CH2)nO]mCH3, O(CH2nOCH3, O(CH2)nNH2, O(CH2)nCH3, O(CH2)nONH2, and O(CH2)nON[(CH2)nCH3)]2, where n and m are from 1 to about 10. In other embodiments, dsRNAs include one of the following at the 2′ position: C1 to C10 lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, 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 iRNA, or a group for improving the pharmacodynamic properties of an iRNA, and other substituents having similar properties. In some embodiments, the modification includes a 2′-methoxyethoxy (2′-O—CH2CH2OCH3, also known as 2′-O-(2-methoxyethyl) or 2′-MOE) (Martin et al., Helv. Chim. Acta, 1995, 78:486-504) i.e., an alkoxy-alkoxy group. Another exemplary modification is 2′-dimethylaminooxyethoxy, i.e., a O(CH2)2ON(CH3)2 group, also known as 2′-DMAOE, as described in examples herein below, and 2′-dimethylaminoethoxyethoxy (also known in the art as 2′-O-dimethylaminoethoxyethyl or 2′-DMAEOE), i.e., 2′-O—CH2—O—CH2—N(CH2)2. Further exemplary modifications include: 5′-Me-2′-F nucleotides, 5′-Me-2′-OMe nucleotides, 5′-Me-2′-deoxynucleotides, (both R and S isomers in these three families); 2′-alkoxyalkyl; and 2′-NMA (N-methylacetamide).


Other modifications include 2′-methoxy (2′-OCH3), 2′-aminopropoxy (2′-OCH2CH2CH2NH2) and 2′-fluoro (2′-F). Similar modifications can also be made at other positions on the RNA of an iRNA, particularly the 3′ position of the sugar on the 3′ terminal nucleotide or in 2′-5′ linked dsRNAs and the 5′ position of 5′ terminal nucleotide. iRNAs can also have sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar. Representative US patents that teach the preparation of such modified sugar structures include, but are not limited to, U.S. Pat. Nos. 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; and 5,700,920, certain of which are commonly owned with the instant application. The entire contents of each of the foregoing are hereby incorporated herein by reference.


An iRNA can also include nucleobase (often referred to in the art simply as “base”) modifications or substitutions. As used herein, “unmodified” or “natural” nucleobases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U). Modified nucleobases include other synthetic and natural nucleobases such as deoxy-thymine (dT), 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 (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl anal other 8-substituted adenines and guanines, 5-halo, particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-daazaadenine and 3-deazaguanine and 3-deazaadenine. Further nucleobases include those disclosed in U.S. Pat. No. 3,687,808, those disclosed in Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijn, P. ed. Wiley-VCH, 2008; those disclosed in The Concise Encyclopedia Of Polymer Science And Engineering, pages 858-859, Kroschwitz, J. L, ed. John Wiley & Sons, 1990, these disclosed by Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613, and those disclosed by Sanghvi, Y S., Chapter 15, dsRNA Research and Applications, pages 289-302, Crooke, S. T. and Lebleu, B., Ed., CRC Press, 1993. Certain of these nucleobases are particularly useful for increasing the binding affinity of the oligomeric compounds featured in the invention. These include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and 0-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine. 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2° C. (Sanghvi, Y. S., Crooke, S. T. and Lebleu, B., Eds., dsRNA Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and are exemplary base substitutions, even more particularly when combined with 2′-O-methoxyethyl sugar modifications.


Representative U.S. patents that teach the preparation of certain of the above noted modified nucleobases as well as other modified nucleobases include, but are not limited to, the above noted U.S. Pat. Nos. 3,687,808, 4,845,205; 5,130,30; 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; 5,594,121, 5,596,091; 5,614,617; 5,681,941; 5,750,692; 6,015,886; 6,147,200; 6,166,197; 6,222,025; 6,235,887; 6,380,368; 6,528,640; 6,639,062; 6,617,438; 7,045,610; 7,427,672; and 7,495,088, the entire contents of each of which are hereby incorporated herein by reference.


The RNA of an iRNA can also be modified to include one or more locked nucleic acids (LNA). A locked nucleic acid is a nucleotide having a modified ribose moiety in which the ribose moiety comprises an extra bridge connecting the 2′ and 4′ carbons. This structure effectively “locks” the ribose in the 3′-endo structural conformation. The addition of locked nucleic acids to siRNAs has been shown to increase siRNA stability in serum, and to reduce off-target effects (Elmen, J. et al., (2005) Nucleic Acids Research 33(1):439-447; Mook, O R. et al., (2007) Mol Canc Ther 6(3):833-843; Grunweller, A. et al., (2003) Nucleic Acids Research 31(12):3185-3193).


In some embodiments, the RNA of an iRNA can also be modified to include one or more bicyclic sugar moieties. A “bicyclic sugar” is a furanosyl ring modified by the bridging of two atoms. A “bicyclic nucleoside” (“BNA”) is a nucleoside having a sugar moiety comprising a bridge connecting two carbon atoms of the sugar ring, thereby forming a bicyclic ring system. In certain embodiments, the bridge connects the 4′-carbon and the 2′-carbon of the sugar ring. Thus, in some embodiments an agent of the invention may include one or more locked nucleic acids (LNA). A locked nucleic acid is a nucleotide having a modified ribose moiety in which the ribose moiety comprises an extra bridge connecting the 2′ and 4′ carbons. In other words, an LNA is a nucleotide comprising a bicyclic sugar moiety comprising a 4′-CH2—O-2′ bridge. This structure effectively “locks” the ribose in the 3′-endo structural conformation. The addition of locked nucleic acids to siRNAs has been shown to increase siRNA stability in serum, and to reduce off-target effects (Elmen, J. et al., (2005) Nucleic Acids Research 33(1):439-447; Mook, O R. et al., (2007) Mol Canc Ther 6(3):833-843; Grunweller, A. et al., (2003) Nucleic Acids Research 31(12):3185-3193). Examples of bicyclic nucleosides for use in the polynucleotides of the invention include without limitation nucleosides comprising a bridge between the 4′ and the 2′ ribosyl ring atoms. In certain embodiments, the antisense polynucleotide agents of the invention include one or more bicyclic nucleosides comprising a 4′ to 2′ bridge. Examples of such 4′ to 2′ bridged bicyclic nucleosides, include but are not limited to 4′-(CH2)—O-2′ (LNA); 4′-(CH2)—S-2′; 4′-(CH2)2—O-2′ (ENA); 4′-CH(CH3)—O-2′ (also referred to as “constrained ethyl” or “cEt”) and 4′-CH(CH2OCH3)—O-2′ (and analogs thereof; see, e.g., U.S. Pat. No. 7,399,845); 4′-C(CH3)(CH3)—O-2′ (and analogs thereof; see e.g., U.S. Pat. No. 8,278,283); 4′-CH2—N(OCH3)-2′ (and analogs thereof; see e.g., U.S. Pat. No. 8,278,425); 4′-CH2—O—N(CH3)-2′ (see, e.g., U.S. Patent Publication No. 2004/0171570); 4′-CH2—N(R)—O-2′, wherein R is H, C1-C12 alkyl, or a protecting group (see, e.g., U.S. Pat. No. 7,427,672); 4′-CH2—C(H)(CH3)-2′ (see, e.g., Chattopadhyaya et al., J. Org. Chem., 2009, 74, 118-134); and 4′-CH2—C(═CH2)-2′ (and analogs thereof; see, e.g., U.S. Pat. No. 8,278,426). The entire contents of each of the foregoing are hereby incorporated herein by reference.


Additional representative U.S. patents and U.S. patenttent Publications that teach the preparation of locked nucleic acid nucleotides include, but are not limited to, the following: U.S. Pat. Nos. 6,268,490; 6,525,191; 6,670,461; 6,770,748; 6,794,499; 6,998,484; 7,053,207; 7,034,133; 7,084,125; 7,399,845; 7,427,672; 7,569,686; 7,741,457; 8,022,193; 8,030,467; 8,278,425; 8,278,426; 8,278,283; US 2008/0039618; and US 2009/0012281, the entire contents of each of which are hereby incorporated herein by reference.


Any of the foregoing bicyclic nucleosides can be prepared having one or more stereochemical sugar configurations including for example α-L-ribofuranose and β-D-ribofuranose (see WO 99/14226).


The RNA of an iRNA can also be modified to include one or more constrained ethyl nucleotides. As used herein, a “constrained ethyl nucleotide” or “cEt” is a locked nucleic acid comprising a bicyclic sugar moiety comprising a 4′-CH(CH3)—O-2′ bridge. In one embodiment, a constrained ethyl nucleotide is in the S conformation referred to herein as “S-cEt.”


An iRNA of the invention may also include one or more “conformationally restricted nucleotides” (“CRN”). CRN are nucleotide analogs with a linker connecting the C2′ and C4′ carbons of ribose or the C3 and —C5′ carbons of ribose. CRN lock the ribose ring into a stable conformation and increase the hybridization affinity to mRNA. The linker is of sufficient length to place the oxygen in an optimal position for stability and affinity resulting in less ribose ring puckering.


Representative publications that teach the preparation of certain of the above noted CRN include, but are not limited to, U.S. Patent Publication No. 2013/0190383; and PCT publication WO 2013/036868, the entire contents of each of which are hereby incorporated herein by reference.


In some embodiments, an iRNA of the invention comprises one or more monomers that are UNA (unlocked nucleic acid) nucleotides. UNA is unlocked acyclic nucleic acid, wherein any of the bonds of the sugar has been removed, forming an unlocked “sugar” residue. In one example, UNA also encompasses monomer with bonds between C1′-C4′ have been removed (i.e. the covalent carbon-oxygen-carbon bond between the C1′ and C4′ carbons). In another example, the C2′-C3′ bond (i.e. the covalent carbon-carbon bond between the C2′ and C3′ carbons) of the sugar has been removed (see Nuc. Acids Symp. Series, 52, 133-134 (2008) and Fluiter et al., Mol. Biosyst., 2009, 10, 1039 hereby incorporated by reference).


Representative U.S. publications that teach the preparation of UNA include, but are not limited to, U.S. Pat. No. 8,314,227; and U.S. Patent Publication Nos. 2013/0096289; 2013/0011922; and 2011/0313020, the entire contents of each of which are hereby incorporated herein by reference.


Potentially stabilizing modifications to the ends of RNA molecules can include N-(acetylaminocaproyl)-4-hydroxyprolinol (Hyp-C6-NHAc), N-(caproyl-4-hydroxyprolinol (Hyp-C6), N-(acetyl-4-hydroxyprolinol (Hyp-NHAc), thymidine-2′-O-deoxythymidine (ether), N-(aminocaproyl)-4-hydroxyprolinol (Hyp-C6-amino), 2-docosanoyl-uridine-3″-phosphate, inverted base dT(idT) and others. Disclosure of this modification can be found in PCT Publication No. WO 2011/005861.


Other modifications of the nucleotides of an iRNA of the invention include a 5′ phosphate or 5′ phosphate mimic, e.g., a 5′-terminal phosphate or phosphate mimic on the antisense strand of an iRNA. Suitable phosphate mimics are disclosed in, for example U.S. Patent Publication No. 2012/0157511, the entire contents of which are incorporated herein by reference.


A. Modified iRNAs Comprising Motifs of the Invention


In certain aspects of the invention, the double stranded RNA agents of the invention include agents with chemical modifications as disclosed, for example, in WO2013/075035, the entire contents of each of which are incorporated herein by reference. WO2013/075035 provides motifs of three identical modifications on three consecutive nucleotides into a sense strand or antisense strand of a dsRNAi agent, particularly at or near the cleavage site. In some embodiments, the sense strand and antisense strand of the dsRNAi agent may otherwise be completely modified. The introduction of these motifs interrupts the modification pattern, if present, of the sense or antisense strand. The dsRNAi agent may be optionally conjugated with a GalNAc derivative ligand, for instance on the sense strand.


More specifically, when the sense strand and antisense strand of the double stranded RNA agent are completely modified to have one or more motifs of three identical modifications on three consecutive nucleotides at or near the cleavage site of at least one strand of a dsRNAi agent, the gene silencing activity of the dsRNAi agent was observed.


Accordingly, the invention provides double stranded RNA agents capable of inhibiting the expression of a target gene (i.e., complement component C3 gene) in vivo. The RNAi agent comprises a sense strand and an antisense strand. Each strand of the RNAi agent may be, for example, 17-30 nucleotides in length, 25-30 nucleotides in length, 27-30 nucleotides in length, 19-25 nucleotides in length, 19-23 nucleotides in length, 19-21 nucleotides in length, 21-25 nucleotides in length, or 21-23 nucleotides in length.


The sense strand and antisense strand typically form a duplex double stranded RNA (“dsRNA”), also referred to herein as “dsRNAi agent.” The duplex region of a dsRNAi agent may be, for example, the duplex region can be 27-30 nucleotide pairs in length, 19-25 nucleotide pairs in length, 19-23 nucleotide pairs in length, 19-21 nucleotide pairs in length, 21-25 nucleotide pairs in length, or 21-23 nucleotide pairs in length. In another example, the duplex region is selected from 19, 20, 21, 22, 23, 24, 25, 26, and 27 nucleotides in length.


In certain embodiments, the dsRNAi agent may contain one or more overhang regions or capping groups at the 3′-end, 5′-end, or both ends of one or both strands. The overhang can be, independently, 1-6 nucleotides in length, for instance 2-6 nucleotides in length, 1-5 nucleotides in length, 2-5 nucleotides in length, 1-4 nucleotides in length, 2-4 nucleotides in length, 1-3 nucleotides in length, 2-3 nucleotides in length, or 1-2 nucleotides in length. In certain embodiments, the overhang regions can include extended overhang regions as provided above. The overhangs can be the result of one strand being longer than the other, or the result of two strands of the same length being staggered. The overhang can form a mismatch with the target mRNA or it can be complementary to the gene sequences being targeted or can be another sequence. The first and second strands can also be joined, e.g., by additional bases to form a hairpin, or by other non-base linkers.


In certain embodiments, the nucleotides in the overhang region of the dsRNAi agent can each independently be a modified or unmodified nucleotide including, but no limited to 2′-sugar modified, such as, 2′-F, 2′-O-methyl, thymidine (T), 2′-O-methoxyethyl-5-methyluridine (Teo), 2′-O-methoxyethyladenosine (Aeo), 2′-O-methoxyethyl-5-methylcytidine (m5Ceo), and any combinations thereof. For example, TT can be an overhang sequence for either end on either strand. The overhang can form a mismatch with the target mRNA or it can be complementary to the gene sequences being targeted or can be another sequence.


The 5′- or 3′-overhangs at the sense strand, antisense strand, or both strands of the dsRNAi agent may be phosphorylated. In some embodiments, the overhang region(s) contains two nucleotides having a phosphorothioate between the two nucleotides, where the two nucleotides can be the same or different. In some embodiments, the overhang is present at the 3′-end of the sense strand, antisense strand, or both strands. In some embodiments, this 3′-overhang is present in the antisense strand. In some embodiments, this 3′-overhang is present in the sense strand.


The dsRNAi agent may contain only a single overhang, which can strengthen the interference activity of the RNAi, without affecting its overall stability. For example, the single-stranded overhang may be located at the 3′-end of the sense strand or, alternatively, at the 3-end of the antisense strand. The RNAi may also have a blunt end, located at the 5′-end of the antisense strand (or the 3′-end of the sense strand) or vice versa. Generally, the antisense strand of the dsRNAi agent has a nucleotide overhang at the 3′-end, and the 5′-end is blunt. While not wishing to be bound by theory, the asymmetric blunt end at the 5′-end of the antisense strand and 3′-end overhang of the antisense strand favor the guide strand loading into RISC process.


In certain embodiments, the dsRNAi agent is a double ended bluntmer of 19 nucleotides in length, wherein the sense strand contains at least one motif of three 2′-F modifications on three consecutive nucleotides at positions 7, 8, 9 from the 5′ end. The antisense strand contains at least one motif of three 2′-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5′ end.


In other embodiments, the dsRNAi agent is a double ended bluntmer of 20 nucleotides in length, wherein the sense strand contains at least one motif of three 2′-F modifications on three consecutive nucleotides at positions 8, 9, 10 from the 5′ end. The antisense strand contains at least one motif of three 2′-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5′ end.


In yet other embodiments, the dsRNAi agent is a double ended bluntmer of 21 nucleotides in length, wherein the sense strand contains at least one motif of three 2′-F modifications on three consecutive nucleotides at positions 9, 10, 11 from the 5′ end. The antisense strand contains at least one motif of three 2′-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5′ end.


In certain embodiments, the dsRNAi agent comprises a 21 nucleotide sense strand and a 23 nucleotide antisense strand, wherein the sense strand contains at least one motif of three 2′-F modifications on three consecutive nucleotides at positions 9, 10, 11 from the 5′ end; the antisense strand contains at least one motif of three 2′-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5′ end, wherein one end of the RNAi agent is blunt, while the other end comprises a 2 nucleotide overhang. Preferably, the 2 nucleotide overhang is at the 3′-end of the antisense strand.


When the 2 nucleotide overhang is at the 3′-end of the antisense strand, there may be two phosphorothioate internucleotide linkages between the terminal three nucleotides, wherein two of the three nucleotides are the overhang nucleotides, and the third nucleotide is a paired nucleotide next to the overhang nucleotide. In one embodiment, the RNAi agent additionally has two phosphorothioate internucleotide linkages between the terminal three nucleotides at both the 5′-end of the sense strand and at the 5′-end of the antisense strand. In certain embodiments, every nucleotide in the sense strand and the antisense strand of the dsRNAi agent, including the nucleotides that are part of the motifs are modified nucleotides. In certain embodiments each residue is independently modified with a 2′-O-methyl or 3′-fluoro, e.g., in an alternating motif. Optionally, the dsRNAi agent further comprises a ligand (preferably GalNAc3).


In certain embodiments, the dsRNAi agent comprises a sense and an antisense strand, wherein the sense strand is 25-30 nucleotide residues in length, wherein starting from the 5′ terminal nucleotide (position 1) positions 1 to 23 of the first strand comprise at least 8 ribonucleotides; the antisense strand is 36-66 nucleotide residues in length and, starting from the 3′ terminal nucleotide, comprises at least 8 ribonucleotides in the positions paired with positions 1-23 of sense strand to form a duplex; wherein at least the 3′ terminal nucleotide of antisense strand is unpaired with sense strand, and up to 6 consecutive 3′ terminal nucleotides are unpaired with sense strand, thereby forming a 3′ single stranded overhang of 1-6 nucleotides; wherein the 5′ terminus of antisense strand comprises from 10-30 consecutive nucleotides which are unpaired with sense strand, thereby forming a 10-30 nucleotide single stranded 5′ overhang; wherein at least the sense strand 5′ terminal and 3′ terminal nucleotides are base paired with nucleotides of antisense strand when sense and antisense strands are aligned for maximum complementarity, thereby forming a substantially duplexed region between sense and antisense strands; and antisense strand is sufficiently complementary to a target RNA along at least 19 ribonucleotides of antisense strand length to reduce target gene expression when the double stranded nucleic acid is introduced into a mammalian cell; and wherein the sense strand contains at least one motif of three 2′-F modifications on three consecutive nucleotides, where at least one of the motifs occurs at or near the cleavage site. The antisense strand contains at least one motif of three 2′-O-methyl modifications on three consecutive nucleotides at or near the cleavage site.


In certain embodiments, the dsRNAi agent comprises sense and antisense strands, wherein the dsRNAi agent comprises a first strand having a length which is at least 25 and at most 29 nucleotides and a second strand having a length which is at most 30 nucleotides with at least one motif of three 2′-O-methyl modifications on three consecutive nucleotides at position 11, 12, 13 from the 5′ end; wherein the 3′ end of the first strand and the 5′ end of the second strand form a blunt end and the second strand is 1-4 nucleotides longer at its 3′ end than the first strand, wherein the duplex region which is at least 25 nucleotides in length, and the second strand is sufficiently complementary to a target mRNA along at least 19 nucleotide of the second strand length to reduce target gene expression when the RNAi agent is introduced into a mammalian cell, and wherein Dicer cleavage of the dsRNAi agent preferentially results in an siRNA comprising the 3′-end of the second strand, thereby reducing expression of the target gene in the mammal. Optionally, the dsRNAi agent further comprises a ligand.


In certain embodiments, the sense strand of the dsRNAi agent contains at least one motif of three identical modifications on three consecutive nucleotides, where one of the motifs occurs at the cleavage site in the sense strand.


In certain embodiments, the antisense strand of the dsRNAi agent can also contain at least one motif of three identical modifications on three consecutive nucleotides, where one of the motifs occurs at or near the cleavage site in the antisense strand.


For a dsRNAi agent having a duplex region of 19-23 nucleotides in length, the cleavage site of the antisense strand is typically around the 10, 11, and 12 positions from the 5′-end. Thus the motifs of three identical modifications may occur at the 9, 10, 11 positions; the 10, 11, 12 positions; the 11, 12, 13 positions; the 12, 13, 14 positions; or the 13, 14, 15 positions of the antisense strand, the count starting from the first nucleotide from the 5′-end of the antisense strand, or, the count starting from the first paired nucleotide within the duplex region from the 5′-end of the antisense strand. The cleavage site in the antisense strand may also change according to the length of the duplex region of the dsRNAi agent from the 5′-end.


The sense strand of the dsRNAi agent may contain at least one motif of three identical modifications on three consecutive nucleotides at the cleavage site of the strand; and the antisense strand may have at least one motif of three identical modifications on three consecutive nucleotides at or near the cleavage site of the strand. When the sense strand and the antisense strand form a dsRNA duplex, the sense strand and the antisense strand can be so aligned that one motif of the three nucleotides on the sense strand and one motif of the three nucleotides on the antisense strand have at least one nucleotide overlap, i.e., at least one of the three nucleotides of the motif in the sense strand forms a base pair with at least one of the three nucleotides of the motif in the antisense strand. Alternatively, at least two nucleotides may overlap, or all three nucleotides may overlap.


In some embodiments, the sense strand of the dsRNAi agent may contain more than one motif of three identical modifications on three consecutive nucleotides. The first motif may occur at or near the cleavage site of the strand and the other motifs may be a wing modification. The term “wing modification” herein refers to a motif occurring at another portion of the strand that is separated from the motif at or near the cleavage site of the same strand. The wing modification is either adjacent to the first motif or is separated by at least one or more nucleotides. When the motifs are immediately adjacent to each other then the chemistries of the motifs are distinct from each other, and when the motifs are separated by one or more nucleotide than the chemistries can be the same or different. Two or more wing modifications may be present. For instance, when two wing modifications are present, each wing modification may occur at one end relative to the first motif which is at or near cleavage site or on either side of the lead motif.


Like the sense strand, the antisense strand of the dsRNAi agent may contain more than one motifs of three identical modifications on three consecutive nucleotides, with at least one of the motifs occurring at or near the cleavage site of the strand. This antisense strand may also contain one or more wing modifications in an alignment similar to the wing modifications that may be present on the sense strand.


In some embodiments, the wing modification on the sense strand or antisense strand of the dsRNAi agent typically does not include the first one or two terminal nucleotides at the 3′-end, 5′-end, or both ends of the strand.


In other embodiments, the wing modification on the sense strand or antisense strand of the dsRNAi agent typically does not include the first one or two paired nucleotides within the duplex region at the 3′-end, 5′-end, or both ends of the strand.


When the sense strand and the antisense strand of the dsRNAi agent each contain at least one wing modification, the wing modifications may fall on the same end of the duplex region, and have an overlap of one, two, or three nucleotides.


When the sense strand and the antisense strand of the dsRNAi agent each contain at least two wing modifications, the sense strand and the antisense strand can be so aligned that two modifications each from one strand fall on one end of the duplex region, having an overlap of one, two, or three nucleotides; two modifications each from one strand fall on the other end of the duplex region, having an overlap of one, two or three nucleotides; two modifications one strand fall on each side of the lead motif, having an overlap of one, two or three nucleotides in the duplex region.


In some embodiments, every nucleotide in the sense strand and antisense strand of the dsRNAi agent, including the nucleotides that are part of the motifs, may be modified. Each nucleotide may be modified with the same or different modification which can include one or more alteration of one or both of the non-linking phosphate oxygens or of one or more of the linking phosphate oxygens; alteration of a constituent of the ribose sugar, e.g., of the 2′-hydroxyl on the ribose sugar; wholesale replacement of the phosphate moiety with “dephospho” linkers; modification or replacement of a naturally occurring base; and replacement or modification of the ribose-phosphate backbone.


As nucleic acids are polymers of subunits, many of the modifications occur at a position which is repeated within a nucleic acid, e.g., a modification of a base, or a phosphate moiety, or a non-linking O of a phosphate moiety. In some cases the modification will occur at all of the subject positions in the nucleic acid but in many cases it will not. By way of example, a modification may only occur at a 3′- or 5′ terminal position, may only occur in a terminal region, e.g., at a position on a terminal nucleotide or in the last 2, 3, 4, 5, or 10 nucleotides of a strand. A modification may occur in a double strand region, a single strand region, or in both. A modification may occur only in the double strand region of an RNA or may only occur in a single strand region of a RNA. For example, a phosphorothioate modification at a non-linking O position may only occur at one or both termini, may only occur in a terminal region, e.g., at a position on a terminal nucleotide or in the last 2, 3, 4, 5, or 10 nucleotides of a strand, or may occur in double strand and single strand regions, particularly at termini. The 5′-end or ends can be phosphorylated.


It may be possible, e.g., to enhance stability, to include particular bases in overhangs, or to include modified nucleotides or nucleotide surrogates, in single strand overhangs, e.g., in a 5′- or 3′-overhang, or in both. For example, it can be desirable to include purine nucleotides in overhangs. In some embodiments all or some of the bases in a 3′- or 5′-overhang may be modified, e.g., with a modification described herein. Modifications can include, e.g., the use of modifications at the 2′ position of the ribose sugar with modifications that are known in the art, e.g., the use of deoxyribonucleotides, 2′-deoxy-2′-fluoro (2′-F) or 2′-O-methyl modified instead of the ribosugar of the nucleobase, and modifications in the phosphate group, e.g., phosphorothioate modifications. Overhangs need not be homologous with the target sequence.


In some embodiments, each residue of the sense strand and antisense strand is independently modified with LNA, CRN, cET, UNA, HNA, CeNA, 2′-methoxyethyl, 2′-O-methyl, 2′-O-allyl, 2′-C-allyl, 2′-deoxy, 2′-hydroxyl, or 2′-fluoro. The strands can contain more than one modification. In one embodiment, each residue of the sense strand and antisense strand is independently modified with 2′-O-methyl or 2′-fluoro.


At least two different modifications are typically present on the sense strand and antisense strand. Those two modifications may be the 2′-O-methyl or 2′-fluoro modifications, or others.


In certain embodiments, the Na or Nb comprise modifications of an alternating pattern. The term “alternating motif” as used herein refers to a motif having one or more modifications, each modification occurring on alternating nucleotides of one strand. The alternating nucleotide may refer to one per every other nucleotide or one per every three nucleotides, or a similar pattern. For example, if A, B and C each represent one type of modification to the nucleotide, the alternating motif can be “ABABABABABAB . . . ,” “AABBAABBAABB . . . ,” “AABAABAABAAB . . . ,” “AAABAAABAAAB . . . ,” “AAABBBAAABBB . . . ,” or “ABCABCABCABC . . . ,” etc.


The type of modifications contained in the alternating motif may be the same or different. For example, if A, B, C, D each represent one type of modification on the nucleotide, the alternating pattern, i.e., modifications on every other nucleotide, may be the same, but each of the sense strand or antisense strand can be selected from several possibilities of modifications within the alternating motif such as “ABABAB . . . ”, “ACACAC . . . ” “BDBDBD . . . ” or “CDCDCD . . . ,” etc.


In some embodiments, the dsRNAi agent of the invention comprises the modification pattern for the alternating motif on the sense strand relative to the modification pattern for the alternating motif on the antisense strand is shifted. The shift may be such that the modified group of nucleotides of the sense strand corresponds to a differently modified group of nucleotides of the antisense strand and vice versa. For example, the sense strand when paired with the antisense strand in the dsRNA duplex, the alternating motif in the sense strand may start with “ABABAB” from 5′ to 3′ of the strand and the alternating motif in the antisense strand may start with “BABABA” from 5′ to 3′ of the strand within the duplex region. As another example, the alternating motif in the sense strand may start with “AABBAABB” from 5′ to 3′ of the strand and the alternating motif in the antisense strand may start with “BBAABBAA” from 5′ to 3′ of the strand within the duplex region, so that there is a complete or partial shift of the modification patterns between the sense strand and the antisense strand.


In some embodiments, the dsRNAi agent comprises the pattern of the alternating motif of 2′-O-methyl modification and 2′-F modification on the sense strand initially has a shift relative to the pattern of the alternating motif of 2′-O-methyl modification and 2′-F modification on the antisense strand initially, i.e., the 2′-O-methyl modified nucleotide on the sense strand base pairs with a 2′-F modified nucleotide on the antisense strand and vice versa. The 1 position of the sense strand may start with the 2′-F modification, and the 1 position of the antisense strand may start with the 2′-O-methyl modification.


The introduction of one or more motifs of three identical modifications on three consecutive nucleotides to the sense strand or antisense strand interrupts the initial modification pattern present in the sense strand or antisense strand. This interruption of the modification pattern of the sense or antisense strand by introducing one or more motifs of three identical modifications on three consecutive nucleotides to the sense or antisense strand may enhance the gene silencing activity against the target gene.


In some embodiments, when the motif of three identical modifications on three consecutive nucleotides is introduced to any of the strands, the modification of the nucleotide next to the motif is a different modification than the modification of the motif. For example, the portion of the sequence containing the motif is “ . . . NaYYYNb . . . ,” where “Y” represents the modification of the motif of three identical modifications on three consecutive nucleotide, and “Na” and “Nb” represent a modification to the nucleotide next to the motif “YYY” that is different than the modification of Y, and where Na and Nb can be the same or different modifications. Alternatively, Na or Nb may be present or absent when there is a wing modification present.


The iRNA may further comprise at least one phosphorothioate or methylphosphonate internucleotide linkage. The phosphorothioate or methylphosphonate internucleotide linkage modification may occur on any nucleotide of the sense strand, antisense strand, or both strands in any position of the strand. For instance, the internucleotide linkage modification may occur on every nucleotide on the sense strand or antisense strand; each internucleotide linkage modification may occur in an alternating pattern on the sense strand or antisense strand; or the sense strand or antisense strand may contain both internucleotide linkage modifications in an alternating pattern. The alternating pattern of the internucleotide linkage modification on the sense strand may be the same or different from the antisense strand, and the alternating pattern of the internucleotide linkage modification on the sense strand may have a shift relative to the alternating pattern of the internucleotide linkage modification on the antisense strand. In one embodiment, a double-stranded RNAi agent comprises 6-8 phosphorothioate internucleotide linkages. In some embodiments, the antisense strand comprises two phosphorothioate internucleotide linkages at the 5′-end and two phosphorothioate internucleotide linkages at the 3′-end, and the sense strand comprises at least two phosphorothioate internucleotide linkages at either the 5′-end or the 3′-end.


In some embodiments, the dsRNAi agent comprises a phosphorothioate or methylphosphonate internucleotide linkage modification in the overhang region. For example, the overhang region may contain two nucleotides having a phosphorothioate or methylphosphonate internucleotide linkage between the two nucleotides. Internucleotide linkage modifications also may be made to link the overhang nucleotides with the terminal paired nucleotides within the duplex region. For example, at least 2, 3, 4, or all the overhang nucleotides may be linked through phosphorothioate or methylphosphonate internucleotide linkage, and optionally, there may be additional phosphorothioate or methylphosphonate internucleotide linkages linking the overhang nucleotide with a paired nucleotide that is next to the overhang nucleotide. For instance, there may be at least two phosphorothioate internucleotide linkages between the terminal three nucleotides, in which two of the three nucleotides are overhang nucleotides, and the third is a paired nucleotide next to the overhang nucleotide. These terminal three nucleotides may be at the 3′-end of the antisense strand, the 3′-end of the sense strand, the 5′-end of the antisense strand, or the 5′ end of the antisense strand.


In some embodiments, the 2-nucleotide overhang is at the 3′-end of the antisense strand, and there are two phosphorothioate internucleotide linkages between the terminal three nucleotides, wherein two of the three nucleotides are the overhang nucleotides, and the third nucleotide is a paired nucleotide next to the overhang nucleotide. Optionally, the dsRNAi agent may additionally have two phosphorothioate internucleotide linkages between the terminal three nucleotides at both the 5′-end of the sense strand and at the 5′-end of the antisense strand.


In one embodiment, the dsRNAi agent comprises mismatch(es) with the target, within the duplex, or combinations thereof. The mismatch may occur in the overhang region or the duplex region. The base pair may be ranked on the basis of their propensity to promote dissociation or melting (e.g., on the free energy of association or dissociation of a particular pairing, the simplest approach is to examine the pairs on an individual pair basis, though next neighbor or similar analysis can also be used). In terms of promoting dissociation: A:U is preferred over G:C; G:U is preferred over G:C; and I:C is preferred over G:C (I=inosine). Mismatches, e.g., non-canonical or other than canonical pairings (as described elsewhere herein) are preferred over canonical (A:T, A:U, G:C) pairings; and pairings which include a universal base are preferred over canonical pairings.


In certain embodiments, the dsRNAi agent comprises at least one of the first 1, 2, 3, 4, or 5 base pairs within the duplex regions from the 5′-end of the antisense strand independently selected from the group of: A:U, G:U, I:C, and mismatched pairs, e.g., non-canonical or other than canonical pairings or pairings which include a universal base, to promote the dissociation of the antisense strand at the 5′-end of the duplex.


In certain embodiments, the nucleotide at the 1 position within the duplex region from the 5′-end in the antisense strand is selected from A, dA, dU, U, and dT. Alternatively, at least one of the first 1, 2, or 3 base pair within the duplex region from the 5′-end of the antisense strand is an AU base pair. For example, the first base pair within the duplex region from the 5′-end of the antisense strand is an AU base pair.


In other embodiments, the nucleotide at the 3′-end of the sense strand is deoxy-thymine (dT) or the nucleotide at the 3′-end of the antisense strand is deoxy-thymine (dT). For example, there is a short sequence of deoxy-thymine nucleotides, for example, two dT nucleotides on the 3′-end of the sense, antisense strand, or both strands.


In certain embodiments, the sense strand sequence may be represented by formula (I):











5′ np-Na-(X X X )i-Nb-Y Y Y -Nb-(Z Z Z )j-Na-Nq






3′ (I)








    • wherein:

    • i and j are each independently 0 or 1;

    • p and q are each independently 0-6;

    • each Na independently represents an oligonucleotide sequence comprising 0-25 modified nucleotides, each sequence comprising at least two differently modified nucleotides;

    • each Nb independently represents an oligonucleotide sequence comprising 0-10 modified nucleotides;

    • each np and nq independently represent an overhang nucleotide;

    • wherein Nb and Y do not have the same modification; and

    • XXX, YYY, and ZZZ each independently represent one motif of three identical modifications on three consecutive nucleotides. Preferably YYY is all 2′-F modified nucleotides.





In some embodiments, the Na or Nb comprises modifications of alternating pattern.


In some embodiments, the YYY motif occurs at or near the cleavage site of the sense strand. For example, when the dsRNAi agent has a duplex region of 17-23 nucleotides in length, the YYY motif can occur at or the vicinity of the cleavage site (e.g.: can occur at positions 6, 7, 8; 7, 8, 9; 8, 9, 10; 9, 10, 11; 10, 11,12; or 11, 12, 13) of the sense strand, the count starting from the first nucleotide, from the 5′-end; or optionally, the count starting at the first paired nucleotide within the duplex region, from the 5′-end.


In one embodiment, i is 1 and j is 0, or i is 0 and j is 1, or both i and j are 1. The sense strand can therefore be represented by the following formulas:











5′ np-Na-YYY-Nb-ZZZ-Na-nq 3′ (Ib);






5′ np-Na-XXX-Nb-YYY-Na-nq 3′ (Ic);



or






5′ np-Na-XXX-Nb-YYY-Nb-ZZZ-Na-nq 3′ (Id).






When the sense strand is represented by formula (Ib), Nb represents an oligonucleotide sequence comprising 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Each Na independently can represent an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.


When the sense strand is represented as formula (Ic), Nb represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Each Na can independently represent an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.


When the sense strand is represented as formula (Id), each Nb independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Preferably, Nb is 0, 1, 2, 3, 4, 5, or 6 Each Na can independently represent an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.


Each of X, Y and Z may be the same or different from each other.


In other embodiments, i is 0 and j is 0, and the sense strand may be represented by the formula:











5′ np-Na-YYY - Na-nq 3′ (Ia).






When the sense strand is represented by formula (Ia), each Na independently can represent an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.


In one embodiment, the antisense strand sequence of the RNAi may be represented by formula (II):









5′ nq′-Na′-(Z′Z′Z)k-Nb′-Y′Y′Y′-Nb′-(X′X′X′)i-N′a-





np′ 3′ (II)








    • wherein:

    • k and 1 are each independently 0 or 1;

    • p′ and q′ are each independently 0-6;

    • each Na′ independently represents an oligonucleotide sequence comprising 0-25 modified nucleotides, each sequence comprising at least two differently modified nucleotides;

    • each Nb′ independently represents an oligonucleotide sequence comprising 0-10 modified nucleotides;

    • each np′ and nq′ independently represent an overhang nucleotide;

    • wherein Nb‘ and Y’ do not have the same modification; and

    • X′X′X′, Y′Y′Y′, and Z′Z′Z′ each independently represent one motif of three identical modifications on three consecutive nucleotides.





In some embodiments, the Na′ or Nb′ comprises modifications of alternating pattern.


The Y′Y′Y′ motif occurs at or near the cleavage site of the antisense strand. For example, when the dsRNAi agent has a duplex region of 17-23 nucleotides in length, the Y′Y′Y′ motif can occur at positions 9, 10, 11; 10, 11, 12; 11, 12, 13; 12, 13, 14; or 13, 14, 15 of the antisense strand, with the count starting from the first nucleotide, from the 5′-end; or optionally, the count starting at the first paired nucleotide within the duplex region, from the 5′-end. Preferably, the Y′Y′Y′ motif occurs at positions 11, 12, 13.


In certain embodiments, Y′Y′Y′ motif is all 2′-OMe modified nucleotides.


In certain embodiments, k is 1 and 1 is 0, or k is 0 and 1 is 1, or both k and 1 are 1.


The antisense strand can therefore be represented by the following formulas:









5′ nq′-Na′-Z′Z′Z′-Nb′-Y′Y′Y′-Na′-np′ 3′ (IIb);





5′ nq′-Na′-Y′Y′Y′-Nb′-X′X′X′-np′ 3′ (IIc);


or





5′ nq′-Na′- Z′Z′Z′-Nb′-Y′Y′Y′-Nb′- X′X′X′-Na′-np′





3′ (IId)






When the antisense strand is represented by formula (IIb), Nb′ represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Each Na′ independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.


When the antisense strand is represented as formula (IIc), Nb′ represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Each Na′ independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.


When the antisense strand is represented as formula (IId), each Nb′ independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Each Na′ independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. Preferably, Nb is 0, 1, 2, 3, 4, 5, or 6.


In other embodiments, k is 0 and 1 is 0 and the antisense strand may be represented by the formula:













5′ np′-Na′-Y′Y′Y′- Na′-nq′ 3′ (Ia).






When the antisense strand is represented as formula (IIa), each Na′ independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. Each of X′, Y′ and Z′ may be the same or different from each other.


Each nucleotide of the sense strand and antisense strand may be independently modified with LNA, CRN, UNA, cEt, HNA, CeNA, 2′-methoxyethyl, 2′-O-methyl, 2′-O-allyl, 2′-C-allyl, 2′-hydroxyl, or 2′-fluoro. For example, each nucleotide of the sense strand and antisense strand is independently modified with 2′-O-methyl or 2′-fluoro. Each X, Y, Z, X′, Y′, and Z′, in particular, may represent a 2′-O-methyl modification or a 2′-fluoro modification.


In some embodiments, the sense strand of the dsRNAi agent may contain YYY motif occurring at 9, 10, and 11 positions of the strand when the duplex region is 21 nt, the count starting from the first nucleotide from the 5′-end, or optionally, the count starting at the first paired nucleotide within the duplex region, from the 5′-end; and Y represents 2′-F modification. The sense strand may additionally contain XXX motif or ZZZ motifs as wing modifications at the opposite end of the duplex region; and XXX and ZZZ each independently represents a 2′-OMe modification or 2′-F modification.


In some embodiments the antisense strand may contain Y′Y′Y′ motif occurring at positions 11, 12, 13 of the strand, the count starting from the first nucleotide from the 5′-end, or optionally, the count starting at the first paired nucleotide within the duplex region, from the 5′-end; and Y′ represents 2′-O-methyl modification. The antisense strand may additionally contain X′X′X′ motif or Z′Z′Z′ motifs as wing modifications at the opposite end of the duplex region; and X′X′X′ and Z′Z′Z′ each independently represents a 2′-OMe modification or 2′-F modification.


The sense strand represented by any one of the above formulas (Ia), (Ib), (Ic), and (Id) forms a duplex with an antisense strand being represented by any one of formulas (IIa), (IIb), (IIc), and (IId), respectively.


Accordingly, the dsRNAi agents for use in the methods of the invention may comprise a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, the iRNA duplex represented by formula (III):









sense:


5′ np -Na-(X X X)i-Nb- Y Y Y -Nb -(Z Z Z)jNa-nq 3′





antisense:


3′ np′-Na′-(X′X′X′)k-Nb′-Y′Y′Y′-Nb′-(Z′Z′Z′)i-Na′-





nq′ 5′ (III)








    • wherein:

    • i, j, k, and l are each independently 0 or 1;

    • p, p′, q, and q′ are each independently 0-6;

    • each Na and Na′ independently represents an oligonucleotide sequence comprising 0-25 modified nucleotides, each sequence comprising at least two differently modified nucleotides;

    • each Nb and Nb′ independently represents an oligonucleotide sequence comprising 0-10 modified nucleotides;

    • wherein each np′, np, nq′, and nq, each of which may or may not be present, independently represents an overhang nucleotide; and

    • XXX, YYY, ZZZ, X′X′X′, Y′Y′Y′, and Z′Z′Z′ each independently represent one motif of three identical modifications on three consecutive nucleotides.





In one embodiment, i is 0 and j is 0; or i is 1 and j is 0; or i is 0 and j is 1; or both i and j are 0; or both i and j are 1. In another embodiment, k is 0 and l is 0; or k is 1 and l is 0; k is 0 and l is 1; or both k and l are 0; or both k and l are 1.


Exemplary combinations of the sense strand and antisense strand forming an iRNA duplex include the formulas below:









5′ np-Na-Y YY-Na-nq 3′





3′ np′-Na′ -Y′Y′Y′ -Na′nq′ 5′


(IIIa)





5′ np -Na -Y Y Y -Nb -Z Z Z -Na-nq 3′





3′ np′-Na′ -Y′Y′Y′-Nb′-Z′Z′Z′-Na′nq′ 5′


(IIIb)





5′ np-Na- X X X -Nb -Y Y Y - Na-nq 3′





3′ np′-Na′-X′X′X′-Nb′-Y′Y′Y′-Na′-nq′ 5′


(IIIc)





5′ np -Na -XXX -Nb-Y Y Y -Nb- Z Z Z -Na-nq 3′





3′ np′-Na′-X′X′X′-Nb′-Y′Y′Y′-Nb′-Z′Z′Z′-Na-nq′ 5′


(IIId)






When the dsRNAi agent is represented by formula (IIIa), each Na independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.


When the dsRNAi agent is represented by formula (IIIb), each Nb independently represents an oligonucleotide sequence comprising 1-10, 1-7, 1-5, or 1-4 modified nucleotides. Each Na independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.


When the dsRNAi agent is represented as formula (IIIc), each Nb, Nb′ independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Each Na independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.


When the dsRNAi agent is represented as formula (IIId), each Nb, Nb′ independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Each Na, Na′ independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. Each of Na, Na′, Nb, and Nb′ independently comprises modifications of alternating pattern.


Each of X, Y, and Z in formulas (III), (IIIa), (IIIb), (IIIc), and (IIId) may be the same or different from each other.


When the dsRNAi agent is represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId), at least one of the Y nucleotides may form a base pair with one of the Y′ nucleotides. Alternatively, at least two of the Y nucleotides form base pairs with the corresponding Y′ nucleotides; or all three of the Y nucleotides all form base pairs with the corresponding Y′ nucleotides.


When the dsRNAi agent is represented by formula (IIIb) or (IIId), at least one of the Z nucleotides may form a base pair with one of the Z′ nucleotides. Alternatively, at least two of the Z nucleotides form base pairs with the corresponding Z′ nucleotides; or all three of the Z nucleotides all form base pairs with the corresponding Z′ nucleotides.


When the dsRNAi agent is represented as formula (IIIc) or (IIId), at least one of the X nucleotides may form a base pair with one of the X′ nucleotides. Alternatively, at least two of the X nucleotides form base pairs with the corresponding X′ nucleotides; or all three of the X nucleotides all form base pairs with the corresponding X′ nucleotides.


In certain embodiments, the modification on the Y nucleotide is different than the modification on the Y′ nucleotide, the modification on the Z nucleotide is different than the modification on the Z′ nucleotide, or the modification on the X nucleotide is different than the modification on the X′ nucleotide.


In certain embodiments, when the dsRNAi agent is represented by formula (IIId), the Na modifications are 2′-O-methyl or 2′-fluoro modifications. In other embodiments, when the RNAi agent is represented by formula (IIId), the Na modifications are 2′-O-methyl or 2′-fluoro modifications and np′>0 and at least one np′ is linked to a neighboring nucleotide a via phosphorothioate linkage. In yet other embodiments, when the RNAi agent is represented by formula (IIId), the Na modifications are 2′-O-methyl or 2′-fluoro modifications, np′>0 and at least one np′ is linked to a neighboring nucleotide via phosphorothioate linkage, and the sense strand is conjugated to one or more GalNAc derivatives attached through a bivalent or trivalent branched linker (described below). In other embodiments, when the RNAi agent is represented by formula (IIId), the Na modifications are 2′-O-methyl or 2′-fluoro modifications, np′>0 and at least one np′ is linked to a neighboring nucleotide via phosphorothioate linkage, the sense strand comprises at least one phosphorothioate linkage, and the sense strand is conjugated to one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.


In some embodiments, when the dsRNAi agent is represented by formula (IIIa), the Na modifications are 2′-O-methyl or 2′-fluoro modifications, np′>0 and at least one np′ is linked to a neighboring nucleotide via phosphorothioate linkage, the sense strand comprises at least one phosphorothioate linkage, and the sense strand is conjugated to one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.


In some embodiments, the dsRNAi agent is a multimer containing at least two duplexes represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId), wherein the duplexes are connected by a linker. The linker can be cleavable or non-cleavable. Optionally, the multimer further comprises a ligand. Each of the duplexes can target the same gene or two different genes; or each of the duplexes can target same gene at two different target sites.


In some embodiments, the dsRNAi agent is a multimer containing three, four, five, six, or more duplexes represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId), wherein the duplexes are connected by a linker. The linker can be cleavable or non-cleavable. Optionally, the multimer further comprises a ligand. Each of the duplexes can target the same gene or two different genes; or each of the duplexes can target same gene at two different target sites.


In one embodiment, two dsRNAi agents represented by at least one of formulas (III), (IIIa), (IIIb), (IIIc), and (IIId) are linked to each other at the 5′ end, and one or both of the 3′ ends, and are optionally conjugated to a ligand. Each of the agents can target the same gene or two different genes; or each of the agents can target same gene at two different target sites.


In certain embodiments, an RNAi agent of the invention may contain a low number of nucleotides containing a 2′-fluoro modification, e.g., 10 or fewer nucleotides with 2′-fluoro modification. For example, the RNAi agent may contain 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or 0 nucleotides with a 2′-fluoro modification. In a specific embodiment, the RNAi agent of the invention contains 10 nucleotides with a 2′-fluoro modification, e.g., 4 nucleotides with a 2′-fluoro modification in the sense strand and 6 nucleotides with a 2′-fluoro modification in the antisense strand. In another specific embodiment, the RNAi agent of the invention contains 6 nucleotides with a 2′-fluoro modification, e.g., 4 nucleotides with a 2′-fluoro modification in the sense strand and 2 nucleotides with a 2′-fluoro modification in the antisense strand.


In other embodiments, an RNAi agent of the invention may contain an ultra low number of nucleotides containing a 2′-fluoro modification, e.g., 2 or fewer nucleotides containing a 2′-fluoro modification. For example, the RNAi agent may contain 2, 1 of 0 nucleotides with a 2′-fluoro modification. In a specific embodiment, the RNAi agent may contain 2 nucleotides with a 2′-fluoro modification, e.g., 0 nucleotides with a 2-fluoro modification in the sense strand and 2 nucleotides with a 2′-fluoro modification in the antisense strand.


Various publications describe multimeric iRNAs that can be used in the methods of the invention. Such publications include WO2007/091269, U.S. Pat. No. 7,858,769, WO2010/141511, WO2007/117686, WO2009/014887, and WO2011/031520 the entire contents of each of which are hereby incorporated herein by reference.


As described in more detail below, the iRNA that contains conjugations of one or more carbohydrate moieties to an iRNA can optimize one or more properties of the iRNA. In many cases, the carbohydrate moiety will be attached to a modified subunit of the iRNA. For example, the ribose sugar of one or more ribonucleotide subunits of a iRNA can be replaced with another moiety, e.g., a non-carbohydrate (preferably cyclic) carrier to which is attached a carbohydrate ligand. A ribonucleotide subunit in which the ribose sugar of the subunit has been so replaced is referred to herein as a ribose replacement modification subunit (RRMS). A cyclic carrier may be a carbocyclic ring system, i.e., all ring atoms are carbon atoms, or a heterocyclic ring system, i.e., one or more ring atoms may be a heteroatom, e.g., nitrogen, oxygen, sulfur. The cyclic carrier may be a monocyclic ring system, or may contain two or more rings, e.g. fused rings. The cyclic carrier may be a fully saturated ring system, or it may contain one or more double bonds.


The ligand may be attached to the polynucleotide via a carrier. The carriers include (i) at least one “backbone attachment point,” preferably two “backbone attachment points” and (ii) at least one “tethering attachment point.” A “backbone attachment point” as used herein refers to a functional group, e.g. a hydroxyl group, or generally, a bond available for, and that is suitable for incorporation of the carrier into the backbone, e.g., the phosphate, or modified phosphate, e.g., sulfur containing, backbone, of a ribonucleic acid. A “tethering attachment point” (TAP) in some embodiments refers to a constituent ring atom of the cyclic carrier, e.g., a carbon atom or a heteroatom (distinct from an atom which provides a backbone attachment point), that connects a selected moiety. The moiety can be, e.g., a carbohydrate, e.g. monosaccharide, disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, or polysaccharide. Optionally, the selected moiety is connected by an intervening tether to the cyclic carrier. Thus, the cyclic carrier will often include a functional group, e.g., an amino group, or generally, provide a bond, that is suitable for incorporation or tethering of another chemical entity, e.g., a ligand to the constituent ring.


The iRNA may be conjugated to a ligand via a carrier, wherein the carrier can be cyclic group or acyclic group; preferably, the cyclic group is selected from pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolane, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuryl, and decalin; preferably, the acyclic group is a serinol backbone or diethanolamine backbone.


In another embodiment of the invention, an iRNA agent comprises a sense strand and an antisense strand, each strand having 14 to 40 nucleotides. The RNAi agent may be represented by formula (L):




embedded image


In formula (L), B1, B2, B3, B1′, B2′, B3′, and B4′ each are independently a nucleotide containing a modification selected from the group consisting of 2′-O-alkyl, 2′-substituted alkoxy, 2′-substituted alkyl, 2′-halo, ENA, and BNA/LNA. In one embodiment, B1, B2, B3, B1′, B2′, B3′, and B4′ each contain 2′-OMe modifications. In one embodiment, B1, B2, B3, B1′, B2′, B3′, and B4′ each contain 2′-OMe or 2′-F modifications. In one embodiment, at least one of B1, B2, B3, B1′, B2′, B3′, and B4′ contain 2′-O—N-methylacetamido (2′-O-NMA) modification.


C1 is a thermally destabilizing nucleotide placed at a site opposite to the seed region of the antisense strand (i.e., at positions 2-8 of the 5′-end of the antisense strand). For example, C1 is at a position of the sense strand that pairs with a nucleotide at positions 2-8 of the 5′-end of the antisense strand. In one example, C1 is at position 15 from the 5′-end of the sense strand. C1 nucleotide bears the thermally destabilizing modification which can include abasic modification; mismatch with the opposing nucleotide in the duplex; and sugar modification such as 2′-deoxy modification or acyclic nucleotide e.g., unlocked nucleic acids (UNA) or glycerol nucleic acid (GNA). In one embodiment, C1 has thermally destabilizing modification selected from the group consisting of: i) mismatch with the opposing nucleotide in the antisense strand; ii) abasic modification selected from the group consisting of:




embedded image


and iii) sugar modification selected from the group consisting of:




embedded image


wherein B is a modified or unmodified nucleobase, R1 and R2 independently are H, halogen, OR3, or alkyl; and R3 is H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl or sugar. In one embodiment, the thermally destabilizing modification in C1 is a mismatch selected from the group consisting of G:G, G:A, G:U, G:T, A:A, A:C, C:C, C:U, C:T, U:U, T:T, and U:T; and optionally, at least one nucleobase in the mismatch pair is a 2′-deoxy nucleobase. In one example, the thermally destabilizing modification in C1 is GNA or




embedded image


T1, T1′, T2′, and T3′ each independently represent a nucleotide comprising a modification providing the nucleotide a steric bulk that is less or equal to the steric bulk of a 2′-OMe modification. A steric bulk refers to the sum of steric effects of a modification. Methods for determining steric effects of a modification of a nucleotide are known to one skilled in the art. The modification can be at the 2′ position of a ribose sugar of the nucleotide, or a modification to a non-ribose nucleotide, acyclic nucleotide, or the backbone of the nucleotide that is similar or equivalent to the 2′ position of the ribose sugar, and provides the nucleotide a steric bulk that is less than or equal to the steric bulk of a 2′-OMe modification. For example, T1, T1′, T2′, and T3′ are each independently selected from DNA, RNA, LNA, 2′-F, and 2′-F-5′-methyl. In one embodiment, T1 is DNA. In one embodiment, T1′ is DNA, RNA or LNA. In one embodiment, T2′ is DNA or RNA. In one embodiment, T3′ is DNA or RNA.

    • n1, n3, and q1 are independently 4 to 15 nucleotides in length.
    • n5, q3, and q7 are independently 1-6 nucleotide(s) in length.
    • n4, q2, and q6 are independently 1-3 nucleotide(s) in length; alternatively, n4 is 0.
    • q5 is independently 0-10 nucleotide(s) in length.
    • n2 and q4 are independently 0-3 nucleotide(s) in length.


Alternatively, n4 is 0-3 nucleotide(s) in length.


In one embodiment, n4 can be 0. In one example, n4 is 0, and q2 and q6 are 1. In another example, n4 is 0, and q2 and q6 are 1, with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand).


In one embodiment, n4, q2, and q6 are each 1.


In one embodiment, n2, n4, q2, q4, and q6 are each 1.


In one embodiment, C1 is at position 14-17 of the 5′-end of the sense strand, when the sense strand is 19-22 nucleotides in length, and n4 is 1. In one embodiment, C1 is at position 15 of the 5′-end of the sense strand


In one embodiment, T3′ starts at position 2 from the 5′ end of the antisense strand. In one example, T3′ is at position 2 from the 5′ end of the antisense strand and q6 is equal to 1.


In one embodiment, T1′ starts at position 14 from the 5′ end of the antisense strand. In one example, T1′ is at position 14 from the 5′ end of the antisense strand and q2 is equal to 1.


In an exemplary embodiment, T3′ starts from position 2 from the 5′ end of the antisense strand and T1′ starts from position 14 from the 5′ end of the antisense strand. In one example, T3′ starts from position 2 from the 5′ end of the antisense strand and q6 is equal to 1 and T1′ starts from position 14 from the 5′ end of the antisense strand and q2 is equal to 1.


In one embodiment, T1′ and T3′ are separated by 11 nucleotides in length (i.e. not counting the T1′ and T3′ nucleotides).


In one embodiment, T1′ is at position 14 from the 5′ end of the antisense strand. In one example, T1′ is at position 14 from the 5′ end of the antisense strand and q2 is equal to 1, and the modification at the 2′ position or positions in a non-ribose, acyclic or backbone that provide less steric bulk than a 2′-OMe ribose.


In one embodiment, T3′ is at position 2 from the 5′ end of the antisense strand. In one example, T3′ is at position 2 from the 5′ end of the antisense strand and q6 is equal to 1, and the modification at the 2′ position or positions in a non-ribose, acyclic or backbone that provide less than or equal to steric bulk than a 2′-OMe ribose.


In one embodiment, T1 is at the cleavage site of the sense strand. In one example, T1 is at position 11 from the 5′ end of the sense strand, when the sense strand is 19-22 nucleotides in length, and n2 is 1. In an exemplary embodiment, T1 is at the cleavage site of the sense strand at position 11 from the 5′ end of the sense strand, when the sense strand is 19-22 nucleotides in length, and n2 is 1, In one embodiment, T2′ starts at position 6 from the 5′ end of the antisense strand. In one example, T2′ is at positions 6-10 from the 5′ end of the antisense strand, and q4 is 1.


In an exemplary embodiment, T1 is at the cleavage site of the sense strand, for instance, at position 11 from the 5′ end of the sense strand, when the sense strand is 19-22 nucleotides in length, and n2 is 1; T1′ is at position 14 from the 5′ end of the antisense strand, and q2 is equal to 1, and the modification to T1′ is at the 2′ position of a ribose sugar or at positions in a non-ribose, acyclic or backbone that provide less steric bulk than a 2′-OMe ribose; T2′ is at positions 6-10 from the 5′ end of the antisense strand, and q4 is 1; and T3′ is at position 2 from the 5′ end of the antisense strand, and q6 is equal to 1, and the modification to T3′ is at the 2′ position or at positions in a non-ribose, acyclic or backbone that provide less than or equal to steric bulk than a 2′-OMe ribose.


In one embodiment, T2′ starts at position 8 from the 5′ end of the antisense strand. In one example, T2′ starts at position 8 from the 5′ end of the antisense strand, and q4 is 2.


In one embodiment, T2′ starts at position 9 from the 5′ end of the antisense strand. In one example, T2′ is at position 9 from the 5′ end of the antisense strand, and q4 is 1.


In one embodiment, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 1, B3′ is 2′-OMe or 2′-F, q5 is 6, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand).


In one embodiment, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 1, B3′ is 2′-OMe or 2′-F, q5 is 6, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand).


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand).


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 6, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 7, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 6, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 7, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand).


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 1, B3′ is 2′-OMe or 2′-F, q5 is 6, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 1, B3′ is 2′-OMe or 2′-F, q5 is 6, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand).


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 5, T2′ is 2′-F, q4 is 1, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; optionally with at least 2 additional TT at the 3′-end of the antisense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 5, T2′ is 2′-F, q4 is 1, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; optionally with at least 2 additional TT at the 3′-end of the antisense strand; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand).


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end).


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand).


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1. In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand).


The RNAi agent can comprise a phosphorus-containing group at the 5′-end of the sense strand or antisense strand. The 5′-end phosphorus-containing group can be 5′-end phosphate (5′-P), 5′-end phosphorothioate (5′-PS), 5′-end phosphorodithioate (5′-PS2), 5′-end vinylphosphonate (5′-VP), 5′-end methylphosphonate (MePhos), or 5′-deoxy-5′-C-malonyl




embedded image


When the 5′-end phosphorus-containing group is 5′-end vinylphosphonate (5′-VP), the 5′-VP can be either 5′-E-VP isomer (i.e., trans-vinylphosphate,




embedded image


5′-Z—VP isomer (i.e., cis-vinylphosphate,




embedded image


or mixtures thereof.


In one embodiment, the RNAi agent comprises a phosphorus-containing group at the 5′-end of the sense strand. In one embodiment, the RNAi agent comprises a phosphorus-containing group at the 5′-end of the antisense strand.


In one embodiment, the RNAi agent comprises a 5′-P. In one embodiment, the RNAi agent comprises a 5′-P in the antisense strand.


In one embodiment, the RNAi agent comprises a 5′-PS. In one embodiment, the RNAi agent comprises a 5′-PS in the antisense strand.


In one embodiment, the RNAi agent comprises a 5′-VP. In one embodiment, the RNAi agent comprises a 5′-VP in the antisense strand. In one embodiment, the RNAi agent comprises a 5′-E-VP in the antisense strand. In one embodiment, the RNAi agent comprises a 5′-Z—VP in the antisense strand.


In one embodiment, the RNAi agent comprises a 5′-PS2. In one embodiment, the RNAi agent comprises a 5′-PS2 in the antisense strand.


In one embodiment, the RNAi agent comprises a 5′-PS2. In one embodiment, the RNAi agent comprises a 5′-deoxy-5′-C-malonyl in the antisense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1. The RNAi agent also comprises a 5′-PS.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1. The RNAi agent also comprises a 5′-P.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1. The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z—VP, or combination thereof.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1. The RNAi agent also comprises a 5′-PS2.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1. The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-P.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z—VP, or combination thereof.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS2.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1. The RNAi agent also comprises a 5′-P.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1. The dsRNA agent also comprises a 5′-PS.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1. The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z—VP, or combination thereof. In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1. The RNAi agent also comprises a 5′-PS2.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1. The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). The RNAi agent also comprises a 5′-P.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). The RNAi agent also comprises a 5′-PS.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z—VP, or combination thereof.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). The RNAi agent also comprises a 5′-PS2.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1. The RNAi agent also comprises a 5′-P.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1. The RNAi agent also comprises a 5′-PS.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1. The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z—VP, or combination thereof.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1. The dsRNAi RNA agent also comprises a 5′-PS2.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1. The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-P.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z—VP, or combination thereof.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS2.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1. The RNAi agent also comprises a 5′-P.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1. The RNAi agent also comprises a 5′-PS.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1. The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z—VP, or combination thereof.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1. The RNAi agent also comprises a 5′-PS2.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1. The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-P.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z—VP, or combination thereof.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS2.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-P and a targeting ligand. In one embodiment, the 5′-P is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS and a targeting ligand. In one embodiment, the 5′-PS is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-VP (e.g., a 5′-E-VP, 5′-Z—VP, or combination thereof), and a targeting ligand.


In one embodiment, the 5′-VP is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS2 and a targeting ligand. In one embodiment, the 5′-PS2 is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl and a targeting ligand. In one embodiment, the 5′-deoxy-5′-C-malonyl is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). The RNAi agent also comprises a 5′-P and a targeting ligand. In one embodiment, the 5′-P is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). The RNAi agent also comprises a 5′-PS and a targeting ligand. In one embodiment, the 5′-PS is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). The RNAi agent also comprises a 5′-VP (e.g., a 5′-E-VP, 5′-Z—VP, or combination thereof) and a targeting ligand. In one embodiment, the 5′-VP is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). The RNAi agent also comprises a 5′-PS2 and a targeting ligand. In one embodiment, the 5′-PS2 is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl and a targeting ligand. In one embodiment, the 5′-deoxy-5′-C-malonyl is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-P and a targeting ligand. In one embodiment, the 5′-P is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS and a targeting ligand. In one embodiment, the 5′-PS is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-VP (e.g., a 5′-E-VP, 5′-Z—VP, or combination thereof) and a targeting ligand. In one embodiment, the 5′-VP is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS2 and a targeting ligand. In one embodiment, the 5′-PS2 is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, T2′ is 2′-F, q4 is 2, B3′ is 2′-OMe or 2′-F, q5 is 5, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl and a targeting ligand. In one embodiment, the 5′-deoxy-5′-C-malonyl is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-P and a targeting ligand. In one embodiment, the 5′-P is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS and a targeting ligand. In one embodiment, the 5′-PS is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-VP (e.g., a 5′-E-VP, 5′-Z—VP, or combination thereof) and a targeting ligand. In one embodiment, the 5′-VP is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-PS2 and a targeting ligand. In one embodiment, the 5′-PS2 is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In one embodiment, B1 is 2′-OMe or 2′-F, n1 is 8, T1 is 2′F, n2 is 3, B2 is 2′-OMe, n3 is 7, n4 is 0, B3 is 2′-OMe, n5 is 3, B1′ is 2′-OMe or 2′-F, q1 is 9, T1′ is 2′-F, q2 is 1, B2′ is 2′-OMe or 2′-F, q3 is 4, q4 is 0, B3′ is 2′-OMe or 2′-F, q5 is 7, T3′ is 2′-F, q6 is 1, B4′ is 2′-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl and a targeting ligand. In one embodiment, the 5′-deoxy-5′-C-malonyl is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand.


In a particular embodiment, an RNAi agent of the present invention comprises:

    • (a) a sense strand having:
      • (i) a length of 21 nucleotides;
      • (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker; and
      • (iii) 2′-F modifications at positions 1, 3, 5, 7, 9 to 11, 13, 17, 19, and 21, and 2′-OMe modifications at positions 2, 4, 6, 8, 12, 14 to 16, 18, and 20 (counting from the 5′ end);
    • and
    • (b) an antisense strand having:
      • (i) a length of 23 nucleotides;
      • (ii) 2′-OMe modifications at positions 1, 3, 5, 9, 11 to 13, 15, 17, 19, 21, and 23, and 2′F modifications at positions 2, 4, 6 to 8, 10, 14, 16, 18, 20, and 22 (counting from the 5′ end); and
      • (iii) phosphorothioate internucleotide linkages between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 (counting from the 5′ end);
    • wherein the dsRNA agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.


In another particular embodiment, an RNAi agent of the present invention comprises:

    • (a) a sense strand having:
      • (i) a length of 21 nucleotides;
      • (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;
      • (iii) 2′-F modifications at positions 1, 3, 5, 7, 9 to 11, 13, 15, 17, 19, and 21, and 2′-OMe modifications at positions 2, 4, 6, 8, 12, 14, 16, 18, and 20 (counting from the 5′ end); and
      • (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);
    • and
    • (b) an antisense strand having:
      • (i) a length of 23 nucleotides;
      • (ii) 2′-OMe modifications at positions 1, 3, 5, 7, 9, 11 to 13, 15, 17, 19, and 21 to 23, and 2′F modifications at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20 (counting from the 5′ end); and
      • (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 (counting from the 5′ end);


        wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.


In another particular embodiment, a RNAi agent of the present invention comprises:

    • (a) a sense strand having:
      • (i) a length of 21 nucleotides;
      • (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;
      • (iii) 2′-OMe modifications at positions 1 to 6, 8, 10, and 12 to 21, 2′-F modifications at positions 7, and 9, and a deoxy-nucleotide (e.g. dT) at position 11 (counting from the 5′ end); and
      • (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);
    • and
    • (b) an antisense strand having:
      • (i) a length of 23 nucleotides;
      • (ii) 2′-OMe modifications at positions 1, 3, 7, 9, 11, 13, 15, 17, and 19 to 23, and 2′-F modifications at positions 2, 4 to 6, 8, 10, 12, 14, 16, and 18 (counting from the 5′ end); and
      • (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 (counting from the 5′ end);


        wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.


In another particular embodiment, a RNAi agent of the present invention comprises:

    • (a) a sense strand having:
      • (i) a length of 21 nucleotides;
      • (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;
      • (iii) 2′-OMe modifications at positions 1 to 6, 8, 10, 12, 14, and 16 to 21, and 2′-F modifications at positions 7, 9, 11, 13, and 15; and
      • (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);
    • and
    • (b) an antisense strand having:
      • (i) a length of 23 nucleotides;
      • (ii) 2′-OMe modifications at positions 1, 5, 7, 9, 11, 13, 15, 17, 19, and 21 to 23, and 2′-F modifications at positions 2 to 4, 6, 8, 10, 12, 14, 16, 18, and 20 (counting from the 5′ end); and
      • (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 (counting from the 5′ end);


        wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.


In another particular embodiment, a RNAi agent of the present invention comprises:

    • (a) a sense strand having:
      • (i) a length of 21 nucleotides;
      • (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;
      • (iii) 2′-OMe modifications at positions 1 to 9, and 12 to 21, and 2′-F modifications at positions 10, and 11; and
      • (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);
    • and
    • (b) an antisense strand having:
      • (i) a length of 23 nucleotides;
      • (ii) 2′-OMe modifications at positions 1, 3, 5, 7, 9, 11 to 13, 15, 17, 19, and 21 to 23, and 2′-F modifications at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20 (counting from the 5′ end); and
      • (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 (counting from the 5′ end);


        wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.


In another particular embodiment, a RNAi agent of the present invention comprises:

    • (a) a sense strand having:
      • (i) a length of 21 nucleotides;
      • (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;
      • (iii) 2′-F modifications at positions 1, 3, 5, 7, 9 to 11, and 13, and 2′-OMe modifications at positions 2, 4, 6, 8, 12, and 14 to 21; and
      • (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);
    • and
    • (b) an antisense strand having:
      • (i) a length of 23 nucleotides;
      • (ii) 2′-OMe modifications at positions 1, 3, 5 to 7, 9, 11 to 13, 15, 17 to 19, and 21 to 23, and 2′-F modifications at positions 2, 4, 8, 10, 14, 16, and 20 (counting from the 5′ end); and (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 (counting from the 5′ end);


        wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.


In another particular embodiment, a RNAi agent of the present invention comprises:

    • (a) a sense strand having:
      • (i) a length of 21 nucleotides;
      • (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;
      • (iii) 2′-OMe modifications at positions 1, 2, 4, 6, 8, 12, 14, 15, 17, and 19 to 21, and 2′-F modifications at positions 3, 5, 7, 9 to 11, 13, 16, and 18; and
      • (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);
    • and
    • (b) an antisense strand having:
      • (i) a length of 25 nucleotides;
      • (ii) 2′-OMe modifications at positions 1, 4, 6, 7, 9, 11 to 13, 15, 17, and 19 to 23, 2′-F modifications at positions 2, 3, 5, 8, 10, 14, 16, and 18, and desoxy-nucleotides (e.g. dT) at positions 24 and 25 (counting from the 5′ end); and
      • (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 (counting from the 5′ end);


        wherein the RNAi agents have a four nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.


In another particular embodiment, a RNAi agent of the present invention comprises:

    • (a) a sense strand having:
      • (i) a length of 21 nucleotides;
      • (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;
      • (iii) 2′-OMe modifications at positions 1 to 6, 8, and 12 to 21, and 2′-F modifications at positions 7, and 9 to 11; and
      • (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);
    • and
    • (b) an antisense strand having:
      • (i) a length of 23 nucleotides;
      • (ii) 2′-OMe modifications at positions 1, 3 to 5, 7, 8, 10 to 13, 15, and 17 to 23, and 2′-F modifications at positions 2, 6, 9, 14, and 16 (counting from the 5′ end); and
      • (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 (counting from the 5′ end);


        wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.


In another particular embodiment, a RNAi agent of the present invention comprises:

    • (a) a sense strand having:
      • (i) a length of 21 nucleotides;
      • (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;
      • (iii) 2′-OMe modifications at positions 1 to 6, 8, and 12 to 21, and 2′-F modifications at positions 7, and 9 to 11; and
      • (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);
    • and
    • (b) an antisense strand having:
      • (i) a length of 23 nucleotides;
      • (ii) 2′-OMe modifications at positions 1, 3 to 5, 7, 10 to 13, 15, and 17 to 23, and 2′-F modifications at positions 2, 6, 8, 9, 14, and 16 (counting from the 5′ end); and
      • (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 (counting from the 5′ end);


        wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.


In another particular embodiment, a RNAi agent of the present invention comprises:

    • (a) a sense strand having:
      • (i) a length of 19 nucleotides;
      • (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;
      • (iii) 2′-OMe modifications at positions 1 to 4, 6, and 10 to 19, and 2′-F modifications at positions 5, and 7 to 9; and
      • (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);
    • and
    • (b) an antisense strand having:
      • (i) a length of 21 nucleotides;
      • (ii) 2′-OMe modifications at positions 1, 3 to 5, 7, 10 to 13, 15, and 17 to 21, and 2′-F modifications at positions 2, 6, 8, 9, 14, and 16 (counting from the 5′ end); and
      • (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 19 and 20, and between nucleotide positions 20 and 21 (counting from the 5′ end);


        wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.


In certain embodiments, the iRNA for use in the methods of the invention is an agent selected from agents listed in any one of Tables 2-7, 15, 18, 20-23, 30, and 31. These agents may further comprise a ligand.


III. iRNAs Conjugated to Ligands


Another modification of the RNA of an iRNA of the invention involves chemically linking to the iRNA one or more ligands, moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the iRNA e.g., into a cell. Such moieties include but are not limited to lipid moieties such as a cholesterol moiety (Letsinger et al., Proc. Natl. Acid. Sci. USA, 1989, 86: 6553-6556). In other embodiments, the ligand is cholic acid (Manoharan et al., Biorg. Med. Chem. Let., 1994, 4:1053-1060), a thioether, e.g., beryl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad. Sci., 1992, 660:306-309; Manoharan et al., Biorg. 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 (Saison-Behmoaras et al., EMBO J, 1991, 10:1111-1118; 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 triethyl-ammonium 1,2-di-O-hexadecyl-rac-glycero-3-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 (Manoharan 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-carbonyloxycholesterol moiety (Crooke et al., J. Pharmacol. Exp. Ther., 1996, 277:923-937).


In certain embodiments, a ligand alters the distribution, targeting, or lifetime of an iRNA agent into which it is incorporated. In preferred embodiments a ligand provides an enhanced affinity for a selected target, e.g., molecule, cell or cell type, compartment, e.g., a cellular or organ compartment, tissue, organ or region of the body, as, e.g., compared to a species absent such a ligand. Preferred ligands do not take part in duplex pairing in a duplexed nucleic acid.


Ligands can include a naturally occurring substance, such as a protein (e.g., human serum albumin (HSA), low-density lipoprotein (LDL), or globulin); carbohydrate (e.g., a dextran, pullulan, chitin, chitosan, inulin, cyclodextrin, N-acetylglucosamine, N-acetylgalactosamine, or hyaluronic acid); or a lipid. The ligand can also be a recombinant or synthetic molecule, such as a synthetic polymer, e.g., a synthetic polyamino acid. Examples of polyamino acids include polyamino acid is a polylysine (PLL), poly L-aspartic acid, poly L-glutamic acid, styrene-maleic acid anhydride copolymer, poly(L-lactide-co-glycolied) copolymer, divinyl ether-maleic anhydride copolymer, N-(2-hydroxypropyl)methacrylamide copolymer (HMPA), polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyurethane, poly(2-ethylacryllic acid), N-isopropylacrylamide polymers, or polyphosphazine. Example of polyamines include: polyethylenimine, polylysine (PLL), spermine, spermidine, polyamine, pseudopeptide-polyamine, peptidomimetic polyamine, dendrimer polyamine, arginine, amidine, protamine, cationic lipid, cationic porphyrin, quaternary salt of a polyamine, or an alpha helical peptide.


Ligands can also include targeting groups, e.g., a cell or tissue targeting agent, e.g., a lectin, glycoprotein, lipid or protein, e.g., an antibody, that binds to a specified cell type such as a kidney cell. A targeting group can be a thyrotropin, melanotropin, lectin, glycoprotein, surfactant protein A, Mucin carbohydrate, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, N-acetyl-glucosamine multivalent mannose, multivalent fucose, glycosylated polyaminoacids, multivalent galactose, transferrin, bisphosphonate, polyglutamate, polyaspartate, a lipid, cholesterol, a steroid, bile acid, folate, vitamin B12, vitamin A, biotin, or an RGD peptide or RGD peptide mimetic. In certain embodiments, the ligand is a multivalent galactose, e.g., an N-acetyl-galactosamine.


Other examples of ligands include dyes, intercalating agents (e.g. acridines), cross-linkers (e.g. psoralene, mitomycin C), porphyrins (TPPC4, texaphyrin, Sapphyrin), polycyclic aromatic hydrocarbons (e.g., phenazine, dihydrophenazine), artificial endonucleases (e.g. EDTA), lipophilic molecules, e.g., cholesterol, cholic acid, adamantane acetic acid, 1-pyrene butyric acid, dihydrotestosterone, 1,3-Bis-O(hexadecyl)glycerol, geranyloxyhexyl group, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, 03-(oleoyl)cholenic acid, dimethoxytrityl, or phenoxazine) and peptide conjugates (e.g., antennapedia peptide, Tat peptide), alkylating agents, phosphate, amino, mercapto, PEG (e.g., PEG-40K), MPEG, [MPEG]2, polyamino, alkyl, substituted alkyl, radiolabeled markers, enzymes, haptens (e.g. biotin), transport/absorption facilitators (e.g., aspirin, vitamin E, folic acid), synthetic ribonucleases (e.g., imidazole, bisimidazole, histamine, imidazole clusters, acridine-imidazole conjugates, Eu3+ complexes of tetraazamacrocycles), dinitrophenyl, HRP, or AP.


Ligands can be proteins, e.g., glycoproteins, or peptides, e.g., molecules having a specific affinity for a co-ligand, or antibodies e.g., an antibody, that binds to a specified cell type such as a hepatic cell. Ligands can also include hormones and hormone receptors. They can also include non-peptidic species, such as lipids, lectins, carbohydrates, vitamins, cofactors, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, N-acetyl-glucosamine multivalent mannose, or multivalent fucose. The ligand can be, for example, a lipopolysaccharide, an activator of p38 MAP kinase, or an activator of NF-κB.


The ligand can be a substance, e.g., a drug, which can increase the uptake of the iRNA agent into the cell, for example, by disrupting the cell's cytoskeleton, e.g., by disrupting the cell's microtubules, microfilaments, or intermediate filaments. The drug can be, for example, taxol, vincristine, vinblastine, cytochalasin, nocodazole, japlakinolide, latrunculin A, phalloidin, swinholide A, indanocine, or myoservin.


In some embodiments, a ligand attached to an iRNA as described herein acts as a pharmacokinetic modulator (PK modulator). PK modulators include lipophiles, bile acids, steroids, phospholipid analogues, peptides, protein binding agents, PEG, vitamins, etc. Exemplary PK modulators include, but are not limited to, cholesterol, fatty acids, cholic acid, lithocholic acid, dialkylglycerides, diacylglyceride, phospholipids, sphingolipids, naproxen, ibuprofen, vitamin E, biotin. Oligonucleotides that comprise a number of phosphorothioate linkages are also known to bind to serum protein, thus short oligonucleotides, e.g., oligonucleotides of about 5 bases, 10 bases, 15 bases, or 20 bases, comprising multiple of phosphorothioate linkages in the backbone are also amenable to the present invention as ligands (e.g. as PK modulating ligands). In addition, aptamers that bind serum components (e.g. serum proteins) are also suitable for use as PK modulating ligands in the embodiments described herein.


Ligand-conjugated iRNAs of the invention may be synthesized by the use of an oligonucleotide that bears a pendant reactive functionality, such as that derived from the attachment of a linking molecule onto the oligonucleotide (described below). This reactive oligonucleotide may be reacted directly with commercially-available ligands, ligands that are synthesized bearing any of a variety of protecting groups, or ligands that have a linking moiety attached thereto.


The oligonucleotides used in the conjugates of the present invention may be conveniently and routinely made through the well-known technique of solid-phase synthesis. Equipment for such synthesis is sold by several vendors including, for example, Applied Biosystems® (Foster City, Calif.). Any other methods for such synthesis known in the art may additionally or alternatively be employed. It is also known to use similar techniques to prepare other oligonucleotides, such as the phosphorothioates and alkylated derivatives.


In the ligand-conjugated iRNAs and ligand-molecule bearing sequence-specific linked nucleosides of the present invention, the oligonucleotides and oligonucleosides may be assembled on a suitable DNA synthesizer utilizing standard nucleotide or nucleoside precursors, or nucleotide or nucleoside conjugate precursors that already bear the linking moiety, ligand-nucleotide or nucleoside-conjugate precursors that already bear the ligand molecule, or non-nucleoside ligand-bearing building blocks.


When using nucleotide-conjugate precursors that already bear a linking moiety, the synthesis of the sequence-specific linked nucleosides is typically completed, and the ligand molecule is then reacted with the linking moiety to form the ligand-conjugated oligonucleotide. In some embodiments, the oligonucleotides or linked nucleosides of the present invention are synthesized by an automated synthesizer using phosphoramidites derived from ligand-nucleoside conjugates in addition to the standard phosphoramidites and non-standard phosphoramidites that are commercially available and routinely used in oligonucleotide synthesis.


A. Lipid Conjugates

In certain embodiments, the ligand or conjugate is a lipid or lipid-based molecule. Such a lipid or lipid-based molecule preferably binds a serum protein, e.g., human serum albumin (HSA). An HSA binding ligand allows for distribution of the conjugate to a target tissue, e.g., a non-kidney target tissue of the body. For example, the target tissue can be the liver, including parenchymal cells of the liver. Other molecules that can bind HSA can also be used as ligands. For example, naproxen or aspirin can be used. A lipid or lipid-based ligand can (a) increase resistance to degradation of the conjugate, (b) increase targeting or transport into a target cell or cell membrane, or (c) can be used to adjust binding to a serum protein, e.g., HSA.


A lipid based ligand can be used to inhibit, e.g., control the binding of the conjugate to a target tissue. For example, a lipid or lipid-based ligand that binds to HSA more strongly will be less likely to be targeted to the kidney and therefore less likely to be cleared from the body. A lipid or lipid-based ligand that binds to HSA less strongly can be used to target the conjugate to the kidney.


In certain embodiments, the lipid based ligand binds HSA. Preferably, it binds HSA with a sufficient affinity such that the conjugate will be preferably distributed to a non-kidney tissue. However, it is preferred that the affinity not be so strong that the HSA-ligand binding cannot be reversed.


In other embodiments, the lipid based ligand binds HSA weakly or not at all, such that the conjugate will be preferably distributed to the kidney. Other moieties that target to kidney cells can also be used in place of, or in addition to, the lipid based ligand.


In another aspect, the ligand is a moiety, e.g., a vitamin, which is taken up by a target cell, e.g., a proliferating cell. These are particularly useful for treating disorders characterized by unwanted cell proliferation, e.g., of the malignant or non-malignant type, e.g., cancer cells. Exemplary vitamins include vitamin A, E, and K. Other exemplary vitamins include are B vitamin, e.g., folic acid, B12, riboflavin, biotin, pyridoxal or other vitamins or nutrients taken up by target cells such as liver cells. Also included are HSA and low density lipoprotein (LDL).


B. Cell Permeation Agents

In another aspect, the ligand is a cell-permeation agent, preferably a helical cell-permeation agent. Preferably, the agent is amphipathic. An exemplary agent is a peptide such as tat or antennopedia. If the agent is a peptide, it can be modified, including a peptidylmimetic, invertomers, non-peptide or pseudo-peptide linkages, and use of D-amino acids. The helical agent is preferably an alpha-helical agent, which preferably has a lipophilic and a lipophobic phase.


The ligand can be a peptide or peptidomimetic. A peptidomimetic (also referred to herein as an oligopeptidomimetic) is a molecule capable of folding into a defined three-dimensional structure similar to a natural peptide. The attachment of peptide and peptidomimetics to iRNA agents can affect pharmacokinetic distribution of the iRNA, such as by enhancing cellular recognition and absorption. The peptide or peptidomimetic moiety can be about 5-50 amino acids long, e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids long.


A peptide or peptidomimetic can be, for example, a cell permeation peptide, cationic peptide, amphipathic peptide, or hydrophobic peptide (e.g., consisting primarily of Tyr, Trp, or Phe). The peptide moiety can be a dendrimer peptide, constrained peptide or crosslinked peptide. In another alternative, the peptide moiety can include a hydrophobic membrane translocation sequence (MTS). An exemplary hydrophobic MTS-containing peptide is RFGF having the amino acid sequence AAVALLPAVLLALLAP (SEQ ID NO: 9). An RFGF analogue (e.g., amino acid sequence AALLPVLLAAP (SEQ ID NO:10) containing a hydrophobic MTS can also be a targeting moiety. The peptide moiety can be a “delivery” peptide, which can carry large polar molecules including peptides, oligonucleotides, and protein across cell membranes. For example, sequences from the HIV Tat protein (GRKKRRQRRRPPQ (SEQ ID NO:11) and the Drosophila Antennapedia protein (RQIKIWFQNRRMKWKK (SEQ ID NO:12) have been found to be capable of functioning as delivery peptides. A peptide or peptidomimetic can be encoded by a random sequence of DNA, such as a peptide identified from a phage-display library, or one-bead-one-compound (OBOC) combinatorial library (Lam et al., Nature, 354:82-84, 1991). Examples of a peptide or peptidomimetic tethered to a dsRNA agent via an incorporated monomer unit for cell targeting purposes is an arginine-glycine-aspartic acid (RGD)-peptide, or RGD mimic. A peptide moiety can range in length from about 5 amino acids to about 40 amino acids. The peptide moieties can have a structural modification, such as to increase stability or direct conformational properties. Any of the structural modifications described below can be utilized.


An RGD peptide for use in the compositions and methods of the invention may be linear or cyclic, and may be modified, e.g., glycosylated or methylated, to facilitate targeting to a specific tissue(s). RGD-containing peptides and peptidiomimemtics may include D-amino acids, as well as synthetic RGD mimics. In addition to RGD, one can use other moieties that target the integrin ligand. Preferred conjugates of this ligand target PECAM-1 or VEGF.


A “cell permeation peptide” is capable of permeating a cell, e.g., a microbial cell, such as a bacterial or fungal cell, or a mammalian cell, such as a human cell. A microbial cell-permeating peptide can be, for example, an α-helical linear peptide (e.g., LL-37 or Ceropin P1), a disulfide bond-containing peptide (e.g., α-defensin, β-defensin or bactenecin), or a peptide containing only one or two dominating amino acids (e.g., PR-39 or indolicidin). A cell permeation peptide can also include a nuclear localization signal (NLS). For example, a cell permeation peptide can be a bipartite amphipathic peptide, such as MPG, which is derived from the fusion peptide domain of HIV-1 gp41 and the NLS of SV40 large T antigen (Simeoni et al., Nucl. Acids Res. 31:2717-2724, 2003).


C. Carbohydrate Conjugates

In some embodiments of the compositions and methods of the invention, an iRNA further comprises a carbohydrate. The carbohydrate conjugated iRNA is advantageous for the in vivo delivery of nucleic acids, as well as compositions suitable for in vivo therapeutic use, as described herein. As used herein, “carbohydrate” refers to a compound which is either a carbohydrate per se made up of one or more monosaccharide units having at least 6 carbon atoms (which can be linear, branched or cyclic) with an oxygen, nitrogen or sulfur atom bonded to each carbon atom; or a compound having as a part thereof a carbohydrate moiety made up of one or more monosaccharide units each having at least six carbon atoms (which can be linear, branched or cyclic), with an oxygen, nitrogen or sulfur atom bonded to each carbon atom. Representative carbohydrates include the sugars (mono-, di-, tri-, and oligosaccharides containing from about 4, 5, 6, 7, 8, or 9 monosaccharide units), and polysaccharides such as starches, glycogen, cellulose and polysaccharide gums. Specific monosaccharides include C5 and above (e.g., C5, C6, C7, or C8) sugars; di- and trisaccharides include sugars having two or three monosaccharide units (e.g., C5, C6, C7, or C8).


In certain embodiments, a carbohydrate conjugate for use in the compositions and methods of the invention is a monosaccharide.


In one embodiment, a carbohydrate conjugate for use in the compositions and methods of the invention is selected from the group consisting of:




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In another embodiment, a carbohydrate conjugate for use in the compositions and methods of the invention is a monosaccharide. In one embodiment, the monosaccharide is an N-acetylgalactosamine, such as




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Another representative carbohydrate conjugate for use in the embodiments described herein includes, but is not limited to,




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(Formula XXXVI), when one of X or Y is an oligonucleotide, the other is a hydrogen.


In certain embodiments of the invention, the GalNAc or GalNAc derivative is attached to an iRNA agent of the invention via a monovalent linker. In some embodiments, the GalNAc or GalNAc derivative is attached to an iRNA agent of the invention via a bivalent linker. In yet other embodiments of the invention, the GalNAc or GalNAc derivative is attached to an iRNA agent of the invention via a trivalent linker.


In one embodiment, the double stranded RNAi agents of the invention comprise one or more GalNAc or GalNAc derivative attached to the iRNA agent. The GalNAc may be attached to any nucleotide via a linker on the sense strand or antsisense strand. The GalNac may be attached to the 5′-end of the sense strand, the 3′ end of the sense strand, the 5′-end of the antisense strand, or the 3′-end of the antisense strand. In one embodiment, the GalNAc is attached to the 3′ end of the sense strand, e.g., via a trivalent linker.


In other embodiments, the double stranded RNAi agents of the invention comprise a plurality (e.g., 2, 3, 4, 5, or 6) GalNAc or GalNAc derivatives, each independently attached to a plurality of nucleotides of the double stranded RNAi agent through a plurality of linkers, e.g., monovalent linkers.


In some embodiments, for example, when the two strands of an iRNA agent of the invention is part of one larger molecule connected by an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′-end of the respective other strand forming a hairpin loop comprising, a plurality of unpaired nucleotides, each unpaired nucleotide within the hairpin loop may independently comprise a GalNAc or GalNAc derivative attached via a monovalent linker.


In some embodiments, the carbohydrate conjugate further comprises one or more additional ligands as described above, such as, but not limited to, a PK modulator or a cell permeation peptide.


Additional carbohydrate conjugates and linkers suitable for use in the present invention include those described in PCT Publication Nos. WO 2014/179620 and WO 2014/179627, the entire contents of each of which are incorporated herein by reference.


D. Linkers

In some embodiments, the conjugate or ligand described herein can be attached to an iRNA oligonucleotide with various linkers that can be cleavable or non-cleavable.


The term “linker” or “linking group” means an organic moiety that connects two parts of a compound, e.g., covalently attaches two parts of a compound. Linkers typically comprise a direct bond or an atom such as oxygen or sulfur, a unit such as NR8, C(O), C(O)NH, SO, SO2, SO2NH or a chain of atoms, such as, but not limited to, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl, alkylarylalkenyl, alkylarylalkynyl, alkenylarylalkyl, alkenylarylalkenyl, alkenylarylalkynyl, alkynylarylalkyl, alkynylarylalkenyl, alkynylarylalkynyl, alkylheteroarylalkyl, alkylheteroarylalkenyl, alkylheteroarylalkynyl, alkenylheteroarylalkyl, alkenylheteroarylalkenyl, alkenylheteroarylalkynyl, alkynylheteroarylalkyl, alkynylheteroarylalkenyl, alkynylheteroarylalkynyl, alkylheterocyclylalkyl, alkylheterocyclylalkenyl, alkylhererocyclylalkynyl, alkenylheterocyclylalkyl, alkenylheterocyclylalkenyl, alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl, alkynylheterocyclylalkenyl, alkynylheterocyclylalkynyl, alkylaryl, alkenylaryl, alkynylaryl, alkylheteroaryl, alkenylheteroaryl, alkynylhereroaryl, which one or more methylenes can be interrupted or terminated by 0, S, S(O), SO2, N(R8), C(O), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted heterocyclic; where R8 is hydrogen, acyl, aliphatic, or substituted aliphatic. In one embodiment, the linker is about 1-24 atoms, 2-24, 3-24, 4-24, 5-24, 6-24, 6-18, 7-18, 8-18, 7-17, 8-17, 6-16, 7-17, or 8-16 atoms.


A cleavable linking group is one which is sufficiently stable outside the cell, but which upon entry into a target cell is cleaved to release the two parts the linker is holding together. In a preferred embodiment, the cleavable linking group is cleaved at least about 10 times, 20, times, 30 times, 40 times, 50 times, 60 times, 70 times, 80 times, 90 times, or more, or at least 100 times faster in a target cell or under a first reference condition (which can, e.g., be selected to mimic or represent intracellular conditions) than in the blood of a subject, or under a second reference condition (which can, e.g., be selected to mimic or represent conditions found in the blood or serum).


Cleavable linking groups are susceptible to cleavage agents, e.g., pH, redox potential, or the presence of degradative molecules. Generally, cleavage agents are more prevalent or found at higher levels or activities inside cells than in serum or blood. Examples of such degradative agents include: redox agents which are selected for particular substrates or which have no substrate specificity, including, e.g., oxidative or reductive enzymes or reductive agents such as mercaptans, present in cells, that can degrade a redox cleavable linking group by reduction; esterases; endosomes or agents that can create an acidic environment, e.g., those that result in a pH of five or lower; enzymes that can hydrolyze or degrade an acid cleavable linking group by acting as a general acid, peptidases (which can be substrate specific), and phosphatases.


A cleavable linkage group, such as a disulfide bond can be susceptible to pH. The pH of human serum is 7.4, while the average intracellular pH is slightly lower, ranging from about 7.1-7.3. Endosomes have a more acidic pH, in the range of 5.5-6.0, and lysosomes have an even more acidic pH at around 5.0. Some linkers will have a cleavable linking group that is cleaved at a preferred pH, thereby releasing a cationic lipid from the ligand inside the cell, or into the desired compartment of the cell.


A linker can include a cleavable linking group that is cleavable by a particular enzyme. The type of cleavable linking group incorporated into a linker can depend on the cell to be targeted. For example, a liver-targeting ligand can be linked to a cationic lipid through a linker that includes an ester group. Liver cells are rich in esterases, and therefore the linker will be cleaved more efficiently in liver cells than in cell types that are not esterase-rich. Other cell-types rich in esterases include cells of the lung, renal cortex, and testis.


Linkers that contain peptide bonds can be used when targeting cell types rich in peptidases, such as liver cells and synoviocytes.


In general, the suitability of a candidate cleavable linking group can be evaluated by testing the ability of a degradative agent (or condition) to cleave the candidate linking group. It will also be desirable to also test the candidate cleavable linking group for the ability to resist cleavage in the blood or when in contact with other non-target tissue. Thus, one can determine the relative susceptibility to cleavage between a first and a second condition, where the first is selected to be indicative of cleavage in a target cell and the second is selected to be indicative of cleavage in other tissues or biological fluids, e.g., blood or serum. The evaluations can be carried out in cell free systems, in cells, in cell culture, in organ or tissue culture, or in whole animals. It can be useful to make initial evaluations in cell-free or culture conditions and to confirm by further evaluations in whole animals. In preferred embodiments, useful candidate compounds are cleaved at least about 2, 4, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 times faster in the cell (or under in vitro conditions selected to mimic intracellular conditions) as compared to blood or serum (or under in vitro conditions selected to mimic extracellular conditions).


i. Redox Cleavable Linking Groups


In certain embodiments, a cleavable linking group is a redox cleavable linking group that is cleaved upon reduction or oxidation. An example of reductively cleavable linking group is a disulphide linking group (—S—S—). To determine if a candidate cleavable linking group is a suitable “reductively cleavable linking group,” or for example is suitable for use with a particular iRNA moiety and particular targeting agent one can look to methods described herein. For example, a candidate can be evaluated by incubation with dithiothreitol (DTT), or other reducing agent using reagents know in the art, which mimic the rate of cleavage which would be observed in a cell, e.g., a target cell. The candidates can also be evaluated under conditions which are selected to mimic blood or serum conditions. In one, candidate compounds are cleaved by at most about 10% in the blood. In other embodiments, useful candidate compounds are degraded at least about 2, 4, 10, 20, 30, 40, 50, 60, 70, 80, 90, or about 100 times faster in the cell (or under in vitro conditions selected to mimic intracellular conditions) as compared to blood (or under in vitro conditions selected to mimic extracellular conditions). The rate of cleavage of candidate compounds can be determined using standard enzyme kinetics assays under conditions chosen to mimic intracellular media and compared to conditions chosen to mimic extracellular media.


ii. Phosphate-Based Cleavable Linking Groups


In other embodiments, a cleavable linker comprises a phosphate-based cleavable linking group. A phosphate-based cleavable linking group is cleaved by agents that degrade or hydrolyze the phosphate group. An example of an agent that cleaves phosphate groups in cells are enzymes such as phosphatases in cells. Examples of phosphate-based linking groups are —O—P(O)(ORk)-O—, —O—P(S)(ORk)-O—, —O—P(S)(SRk)-O—, —S—P(O)(ORk)-O—, —O—P(O)(ORk)-S—, —S—P(O)(ORk)-S—, —O—P(S)(ORk)-S—, —S—P(S)(ORk)-O—, —O—P(O)(Rk)-O—, —O—P(S)(Rk)-O—, —S—P(O)(Rk)-O—, —S—P(S)(Rk)-O—, —S—P(O)(Rk)-S—, —O—P(S)(Rk)-S—. Preferred embodiments are —O—P(O)(OH)—O—, —O—P(S)(OH)—O—, —O—P(S)(SH)—O—, —S—P(O)(OH)—O—, —O—P(O)(OH)—S—, —S—P(O)(OH)—S—, —O—P(S)(OH)—S—, —S—P(S)(OH)—O—, —O—P(O)(H)—O—, —O—P(S)(H)—O—, —S—P(O)(H)—O, —S—P(S)(H)—O—, —S—P(O)(H)—S—, and —O—P(S)(H)—S—. A preferred embodiment is —O—P(O)(OH)—O—. These candidates can be evaluated using methods analogous to those described above.


iii. Acid Cleavable Linking Groups


In other embodiments, a cleavable linker comprises an acid cleavable linking group. An acid cleavable linking group is a linking group that is cleaved under acidic conditions. In preferred embodiments acid cleavable linking groups are cleaved in an acidic environment with a pH of about 6.5 or lower (e.g., about 6.0, 5.5, 5.0, or lower), or by agents such as enzymes that can act as a general acid. In a cell, specific low pH organelles, such as endosomes and lysosomes can provide a cleaving environment for acid cleavable linking groups. Examples of acid cleavable linking groups include but are not limited to hydrazones, esters, and esters of amino acids. Acid cleavable groups can have the general formula —C═NN—, C(O)O, or —OC(O). A preferred embodiment is when the carbon attached to the oxygen of the ester (the alkoxy group) is an aryl group, substituted alkyl group, or tertiary alkyl group such as dimethyl pentyl or t-butyl. These candidates can be evaluated using methods analogous to those described above.


iv. Ester-Based Linking Groups


In other embodiments, a cleavable linker comprises an ester-based cleavable linking group. An ester-based cleavable linking group is cleaved by enzymes such as esterases and amidases in cells. Examples of ester-based cleavable linking groups include, but are not limited to, esters of alkylene, alkenylene and alkynylene groups. Ester cleavable linking groups have the general formula —C(O)O—, or —OC(O)—. These candidates can be evaluated using methods analogous to those described above.


v. Peptide-Based Cleaving Groups


In yet other embodiments, a cleavable linker comprises a peptide-based cleavable linking group. A peptide-based cleavable linking group is cleaved by enzymes such as peptidases and proteases in cells. Peptide-based cleavable linking groups are peptide bonds formed between amino acids to yield oligopeptides (e.g., dipeptides, tripeptides etc.) and polypeptides. Peptide-based cleavable groups do not include the amide group (—C(O)NH—). The amide group can be formed between any alkylene, alkenylene or alkynelene. A peptide bond is a special type of amide bond formed between amino acids to yield peptides and proteins. The peptide based cleavage group is generally limited to the peptide bond (i.e., the amide bond) formed between amino acids yielding peptides and proteins and does not include the entire amide functional group. Peptide-based cleavable linking groups have the general formula —NHCHRAC(O)NHCHRBC(O)—, where RA and RB are the R groups of the two adjacent amino acids. These candidates can be evaluated using methods analogous to those described above.


In some embodiments, an iRNA of the invention is conjugated to a carbohydrate through a linker. Non-limiting examples of iRNA carbohydrate conjugates with linkers of the compositions and methods of the invention include, but are not limited to,




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when one of X or Y is an oligonucleotide, the other is a hydrogen.


In certain embodiments of the compositions and methods of the invention, a ligand is one or more “GalNAc” (N-acetylgalactosamine) derivatives attached through a bivalent or trivalent branched linker.


In one embodiment, a dsRNA of the invention is conjugated to a bivalent or trivalent branched linker selected from the group of structures shown in any of formula (XLV)-(XLVI):




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wherein:

    • q2A, q2B, q3A, q3B, q4A, q4B, q5A, q5B and q5C represent independently for each occurrence 0-20 and wherein the repeating unit can be the same or different;
    • P2A, P2B, P3A, P3B, P4A, P4B, P5A, P5B, P5C, T2A, T2B, T3A, T3B, T4A, T4B, T4A, T5B, T5C are each independently for each occurrence absent, CO, NH, O, S, OC(O), NHC(O), CH2, CH2NH or CH2O;
    • Q2A, Q2B, Q3A, Q3B, Q4A, Q4B, Q5A, Q5B, Q5C are independently for each occurrence absent, alkylene, substituted alkylene wherein one or more methylenes can be interrupted or terminated by one or more of O, S, S(O), SO2, N(RN), C(R′)═C(R″), C≡C or C(O);
    • R2A, R2B, R3A, R3B, R4A, R4B, R5A, R5B, R5C are each independently for each occurrence absent, NH, O, S, CH2, C(O)O, C(O)NH, NHCH(Ra)C(O), —C(O)—CH(Ra)—NH—, CO, CH═N—O, N




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or heterocyclyl;

    • L2A, L2B, L3A, L3B, L4A, L4B L5A, L5B and L5C represent the ligand; i.e. each independently for each occurrence a monosaccharide (such as GalNAc), disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, or polysaccharide; and Ra is H or amino acid side chain. Trivalent conjugating GalNAc derivatives are particularly useful for use with RNAi agents for inhibiting the expression of a target gene, such as those of formula (XLIX):




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wherein L5A, L5B and L5c represent a monosaccharide, such as GalNAc derivative.


Examples of suitable bivalent and trivalent branched linker groups conjugating GalNAc derivatives include, but are not limited to, the structures recited above as formulas II, VII, XI, X, and XIII.


Representative U.S. patents that teach the preparation of RNA conjugates include, but are not limited to, 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; 5,591,584; 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; 5,688,941; 6,294,664; 6,320,017; 6,576,752; 6,783,931; 6,900,297; 7,037,646; and 8,106,022, the entire contents of each of which are hereby incorporated herein by reference.


It is not necessary for all positions in a given compound to be uniformly modified, and in fact more than one of the aforementioned modifications can be incorporated in a single compound or even at a single nucleoside within an iRNA. The present invention also includes iRNA compounds that are chimeric compounds.


“Chimeric” iRNA compounds or “chimeras,” in the context of this invention, are iRNA compounds, preferably dsRNAi agents, that contain two or more chemically distinct regions, each made up of at least one monomer unit, i.e., a nucleotide in the case of a dsRNA compound. These iRNAs typically contain at least one region wherein the RNA is modified so as to confer upon the iRNA increased resistance to nuclease degradation, increased cellular uptake, or increased binding affinity for the target nucleic acid. An additional region of the iRNA can serve as a substrate for enzymes capable of cleaving RNA:DNA or RNA:RNA hybrids. By way of example, RNase H is a cellular endonuclease which cleaves the RNA strand of an RNA:DNA duplex. Activation of RNase H, therefore, results in cleavage of the RNA target, thereby greatly enhancing the efficiency of iRNA inhibition of gene expression. Consequently, comparable results can often be obtained with shorter iRNAs when chimeric dsRNAs are used, compared to phosphorothioate deoxy dsRNAs hybridizing to the same target region. Cleavage of the RNA target can be routinely detected by gel electrophoresis and, if necessary, associated nucleic acid hybridization techniques known in the art.


In certain instances, the RNA of an iRNA can be modified by a non-ligand group. A number of non-ligand molecules have been conjugated to iRNAs in order to enhance the activity, cellular distribution or cellular uptake of the iRNA, and procedures for performing such conjugations are available in the scientific literature. Such non-ligand moieties have included lipid moieties, such as cholesterol (Kubo, T. et al., Biochem. Biophys. Res. Comm., 2007, 365(1):54-61; Letsinger et al., Proc. Natl. Acad. Sci. USA, 1989, 86:6553), cholic acid (Manoharan et al., Bioorg. Med. Chem. Lett., 1994, 4:1053), a thioether, e.g., hexyl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad. Sci., 1992, 660:306; Manoharan et al., Bioorg. Med. Chem. Let., 1993, 3:2765), a thiocholesterol (Oberhauser et al., Nucl. Acids Res., 1992, 20:533), an aliphatic chain, e.g., dodecandiol or undecyl residues (Saison-Behmoaras et al., EMBO J., 1991, 10:111; Kabanov et al., FEBS Lett., 1990, 259:327; Svinarchuk et al., Biochimie, 1993, 75:49), 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; Shea et al., Nucl. Acids Res., 1990, 18:3777), a polyamine or a polyethylene glycol chain (Manoharan et al., Nucleosides & Nucleotides, 1995, 14:969), or adamantane acetic acid (Manoharan et al., Tetrahedron Lett., 1995, 36:3651), a palmityl moiety (Mishra et al., Biochim. Biophys. Acta, 1995, 1264:229), or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke et al., J. Pharmacol. Exp. Ther., 1996, 277:923). Representative United States patents that teach the preparation of such RNA conjugates have been listed above. Typical conjugation protocols involve the synthesis of RNAs bearing an aminolinker at one or more positions of the sequence. The amino group is then reacted with the molecule being conjugated using appropriate coupling or activating reagents. The conjugation reaction can be performed either with the RNA still bound to the solid support or following cleavage of the RNA, in solution phase. Purification of the RNA conjugate by HPLC typically affords the pure conjugate.


IV. Delivery of an iRNA of the Invention


The delivery of an iRNA of the invention to a cell e.g., a cell within a subject, such as a human subject (e.g., a subject in need thereof, such as a subject susceptible to or diagnosed with a complement component C3-associated disorder, e.g., hemolysis) can be achieved in a number of different ways. For example, delivery may be performed by contacting a cell with an iRNA of the invention either in vitro or in vivo. In vivo delivery may also be performed directly by administering a composition comprising an iRNA, e.g., a dsRNA, to a subject. Alternatively, in vivo delivery may be performed indirectly by administering one or more vectors that encode and direct the expression of the iRNA. These alternatives are discussed further below.


In general, any method of delivering a nucleic acid molecule (in vitro or in vivo) can be adapted for use with an iRNA of the invention (see e.g., Akhtar S. and Julian R L. (1992) Trends Cell. Biol. 2(5):139-144 and WO94/02595, which are incorporated herein by reference in their entireties). For in vivo delivery, factors to consider in order to deliver an iRNA molecule include, for example, biological stability of the delivered molecule, prevention of non-specific effects, and accumulation of the delivered molecule in the target tissue. RNA interference has also shown success with local delivery to the CNS by direct injection (Dorn, G., et al. (2004) Nucleic Acids 32:e49; Tan, P H., et al (2005) Gene Ther. 12:59-66; Makimura, H., et al (2002) BMC Neurosci. 3:18; Shishkina, G T., et al (2004) Neuroscience 129:521-528; Thakker, E R., et al (2004) Proc. Natl. Acad. Sci. U.S.A. 101:17270-17275; Akaneya, Y., et al (2005) J. Neurophysiol. 93:594-602). Modification of the RNA or the pharmaceutical carrier can also permit targeting of the iRNA to the target tissue and avoid undesirable off-target effects. iRNA molecules can be modified by chemical conjugation to lipophilic groups such as cholesterol to enhance cellular uptake and prevent degradation. For example, an iRNA directed against ApoB conjugated to a lipophilic cholesterol moiety was injected systemically into mice and resulted in knockdown of apoB mRNA in both the liver and jejunum (Soutschek, J., et al (2004) Nature 432:173-178).


In an alternative embodiment, the iRNA can be delivered using drug delivery systems such as a nanoparticle, a dendrimer, a polymer, liposomes, or a cationic delivery system. Positively charged cationic delivery systems facilitate binding of an iRNA molecule (negatively charged) and also enhance interactions at the negatively charged cell membrane to permit efficient uptake of an iRNA by the cell. Cationic lipids, dendrimers, or polymers can either be bound to an iRNA, or induced to form a vesicle or micelle (see e.g., Kim S H, et al (2008) Journal of Controlled Release 129(2):107-116) that encases an iRNA. The formation of vesicles or micelles further prevents degradation of the iRNA when administered systemically. Methods for making and administering cationic-iRNA complexes are well within the abilities of one skilled in the art (see e.g., Sorensen, D R, et al (2003) J. Mol. Biol 327:761-766; Verma, U N, et al (2003) Clin. Cancer Res. 9:1291-1300; Arnold, A S et al (2007) J. Hypertens. 25:197-205, which are incorporated herein by reference in their entirety). Some non-limiting examples of drug delivery systems useful for systemic delivery of iRNAs include DOTAP (Sorensen, D R., et al (2003), supra; Verma, U N, et al (2003), supra), “solid nucleic acid lipid particles” (Zimmermann, T S, et al (2006) Nature 441:111-114), cardiolipin (Chien, P Y, et al (2005) Cancer Gene Ther. 12:321-328; Pal, A, et al (2005) Int J. Oncol. 26:1087-1091), polyethyleneimine (Bonnet M E, et al (2008) Pharm. Res. August 16 Epub ahead of print; Aigner, A. (2006) J. Biomed. Biotechnol. 71659), Arg-Gly-Asp (RGD) peptides (Liu, S. (2006) Mol. Pharm. 3:472-487), and polyamidoamines (Tomalia, D A, et al (2007) Biochem. Soc. Trans. 35:61-67; Yoo, H., et al (1999) Pharm. Res. 16:1799-1804). In some embodiments, an iRNA forms a complex with cyclodextrin for systemic administration. Methods for administration and pharmaceutical compositions of iRNAs and cyclodextrins can be found in U.S. Pat. No. 7,427,605, which is herein incorporated by reference in its entirety.


A. Vector Encoded iRNAs of the Invention


iRNA targeting the complement component C3 gene can be expressed from transcription units inserted into DNA or RNA vectors (see, e.g., Couture, A, et al., TIG. (1996), 12:5-10; Skillern, A, et al., International PCT Publication No. WO 00/22113, Conrad, International PCT Publication No. WO 00/22114, and Conrad, U.S. Pat. No. 6,054,299). Expression can be transient (on the order of hours to weeks) or sustained (weeks to months or longer), depending upon the specific construct used and the target tissue or cell type. These transgenes can be introduced as a linear construct, a circular plasmid, or a viral vector, which can be an integrating or non-integrating vector. The transgene can also be constructed to permit it to be inherited as an extrachromosomal plasmid (Gassmann, et al., Proc. Natl. Acad. Sci. USA (1995) 92:1292).


Viral vector systems which can be utilized with the methods and compositions described herein include, but are not limited to, (a) adenovirus vectors; (b) retrovirus vectors, including but not limited to lentiviral vectors, moloney murine leukemia virus, etc.; (c) adeno-associated virus vectors; (d) herpes simplex virus vectors; (e) SV 40 vectors; (f) polyoma virus vectors; (g) papilloma virus vectors; (h) picornavirus vectors; (i) pox virus vectors such as an orthopox, e.g., vaccinia virus vectors or avipox, e.g. canary pox or fowl pox; and (j) a helper-dependent or gutless adenovirus. Replication-defective viruses can also be advantageous. Different vectors will or will not become incorporated into the cells' genome. The constructs can include viral sequences for transfection, if desired. Alternatively, the construct can be incorporated into vectors capable of episomal replication, e.g. EPV and EBV vectors. Constructs for the recombinant expression of an iRNA will generally require regulatory elements, e.g., promoters, enhancers, etc., to ensure the expression of the iRNA in target cells. Other aspects to consider for vectors and constructs are known in the art.


V. Pharmaceutical Compositions of the Invention

The present invention also includes pharmaceutical compositions and formulations which include the iRNAs of the invention. In one embodiment, provided herein are pharmaceutical compositions containing an iRNA, as described herein, and a pharmaceutically acceptable carrier. The pharmaceutical compositions containing the iRNA are useful for preventing or treating a complement component C3-associated disorder, e.g., hemolysis. Such pharmaceutical compositions are formulated based on the mode of delivery. One example is compositions that are formulated for systemic administration via parenteral delivery, e.g., by subcutaneous (SC), intramuscular (IM), or intravenous (IV) delivery. The pharmaceutical compositions of the invention may be administered in dosages sufficient to inhibit expression of a complement component C3 gene.


In some embodiments, the pharmaceutical compositions of the invention are sterile. In another embodiment, the pharmaceutical compositions of the invention are pyrogen free.


The pharmaceutical compositions of the invention may be administered in dosages sufficient to inhibit expression of a complement component C3 gene. In general, a suitable dose of an iRNA of the invention will be in the range of about 0.001 to about 200.0 milligrams per kilogram body weight of the recipient per day, generally in the range of about 1 to 50 mg per kilogram body weight per day. Typically, a suitable dose of an iRNA of the invention will be in the range of about 0.1 mg/kg to about 5.0 mg/kg, preferably about 0.3 mg/kg and about 3.0 mg/kg. A repeat-dose regimen may include administration of a therapeutic amount of iRNA on a regular basis, such as every month, once every 3-6 months, or once a year. In certain embodiments, the iRNA is administered about once per month to about once per six months.


After an initial treatment regimen, the treatments can be administered on a less frequent basis. Duration of treatment can be determined based on the severity of disease.


In other embodiments, a single dose of the pharmaceutical compositions can be long lasting, such that doses are administered at not more than 1, 2, 3, or 4 month intervals. In some embodiments of the invention, a single dose of the pharmaceutical compositions of the invention is administered about once per month. In other embodiments of the invention, a single dose of the pharmaceutical compositions of the invention is administered quarterly (i.e., about every three months). In other embodiments of the invention, a single dose of the pharmaceutical compositions of the invention is administered twice per year (i.e., about once every six months).


The skilled artisan will appreciate that certain factors can influence the dosage and timing required to effectively treat a subject, including but not limited to mutations present in the subject, previous treatments, the general health or age of the subject, and other diseases present. Moreover, treatment of a subject with a prophylactically or therapeutically effective amount, as appropriate, of a composition can include a single treatment or a series of treatments.


The iRNA can be delivered in a manner to target a particular tissue (e.g., hepatocytes).


Pharmaceutical compositions of the present invention include, but are not limited to, solutions, emulsions, and liposome-containing formulations. These compositions can be generated from a variety of components that include, but are not limited to, preformed liquids, self-emulsifying solids, and self-emulsifying semisolids. Formulations include those that target the liver.


The pharmaceutical formulations of the present invention, which can conveniently be presented in unit dosage form, can be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing into association the active ingredients with the pharmaceutical carrier(s) or excipient(s). In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers.


A. Additional Formulations

i. Emulsions


The compositions of the present invention can be prepared and formulated as emulsions. Emulsions are typically heterogeneous systems of one liquid dispersed in another in the form of droplets usually exceeding 0.1 μm in diameter (see e.g., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, LV., Popovich N G., and Ansel H C., 2004, Lippincott Williams & Wilkins (8th ed.), New York, NY; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., Volume 1, p. 245; Block in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 2, p. 335; Higuchi et al., in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1985, p. 301). Emulsions are often biphasic systems comprising two immiscible liquid phases intimately mixed and dispersed with each other. In general, emulsions can be of either the water-in-oil (w/o) or the oil-in-water (o/w) variety. When an aqueous phase is finely divided into and dispersed as minute droplets into a bulk oily phase, the resulting composition is called a water-in-oil (w/o) emulsion. Alternatively, when an oily phase is finely divided into and dispersed as minute droplets into a bulk aqueous phase, the resulting composition is called an oil-in-water (o/w) emulsion. Emulsions can contain additional components in addition to the dispersed phases, and the active drug which can be present as a solution either in the aqueous phase, oily phase or itself as a separate phase. Pharmaceutical excipients such as emulsifiers, stabilizers, dyes, and anti-oxidants can also be present in emulsions as needed. Pharmaceutical emulsions can also be multiple emulsions that are comprised of more than two phases such as, for example, in the case of oil-in-water-in-oil (o/w/o) and water-in-oil-in-water (w/o/w) emulsions. Such complex formulations often provide certain advantages that simple binary emulsions do not. Multiple emulsions in which individual oil droplets of an o/w emulsion enclose small water droplets constitute a w/o/w emulsion. Likewise a system of oil droplets enclosed in globules of water stabilized in an oily continuous phase provides an o/w/o emulsion.


Emulsions are characterized by little or no thermodynamic stability. Often, the dispersed or discontinuous phase of the emulsion is well dispersed into the external or continuous phase and maintained in this form through the means of emulsifiers or the viscosity of the formulation. Other means of stabilizing emulsions entail the use of emulsifiers that can be incorporated into either phase of the emulsion. Emulsifiers can broadly be classified into four categories: synthetic surfactants, naturally occurring emulsifiers, absorption bases, and finely dispersed solids (see e.g., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, LV., Popovich N G., and Ansel H C., 2004, Lippincott Williams & Wilkins (8th ed.), New York, NY; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199).


Synthetic surfactants, also known as surface active agents, have found wide applicability in the formulation of emulsions and have been reviewed in the literature (see e.g., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, LV., Popovich N G., and Ansel H C., 2004, Lippincott Williams & Wilkins (8th ed.), New York, NY; Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 285; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), Marcel Dekker, Inc., New York, N.Y., 1988, volume 1, p. 199). Surfactants are typically amphiphilic and comprise a hydrophilic and a hydrophobic portion. The ratio of the hydrophilic to the hydrophobic nature of the surfactant has been termed the hydrophile/lipophile balance (HLB) and is a valuable tool in categorizing and selecting surfactants in the preparation of formulations. Surfactants can be classified into different classes based on the nature of the hydrophilic group: nonionic, anionic, cationic, and amphoteric (see e.g., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, LV., Popovich N G., and Ansel H C., 2004, Lippincott Williams & Wilkins (8th ed.), New York, NY Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 285).


A large variety of non-emulsifying materials are also included in emulsion formulations and contribute to the properties of emulsions. These include fats, oils, waxes, fatty acids, fatty alcohols, fatty esters, humectants, hydrophilic colloids, preservatives, and antioxidants (Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 335; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199).


The application of emulsion formulations via dermatological, oral, and parenteral routes, and methods for their manufacture have been reviewed in the literature (see e.g., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, LV., Popovich N G., and Ansel H C., 2004, Lippincott Williams & Wilkins (8th ed.), New York, NY; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199).


ii. Microemulsions


In one embodiment of the present invention, the compositions of iRNAs and nucleic acids are formulated as microemulsions. A microemulsion can be defined as a system of water, oil, and amphiphile which is a single optically isotropic and thermodynamically stable liquid solution (see e.g., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, LV., Popovich N G., and Ansel H C., 2004, Lippincott Williams & Wilkins (8th ed.), New York, NY; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245). Typically microemulsions are systems that are prepared by first dispersing an oil in an aqueous surfactant solution and then adding a sufficient amount of a fourth component, generally an intermediate chain-length alcohol to form a transparent system. Therefore, microemulsions have also been described as thermodynamically stable, isotropically clear dispersions of two immiscible liquids that are stabilized by interfacial films of surface-active molecules (Leung and Shah, in: Controlled Release of Drugs: Polymers and Aggregate Systems, Rosoff, M., Ed., 1989, VCH Publishers, New York, pages 185-215).


iii. Microparticles


An iRNA of the invention may be incorporated into a particle, e.g., a microparticle. Microparticles can be produced by spray-drying, but may also be produced by other methods including lyophilization, evaporation, fluid bed drying, vacuum drying, or a combination of these techniques.


iv. Penetration Enhancers


In one embodiment, the present invention employs various penetration enhancers to effect the efficient delivery of nucleic acids, particularly iRNAs, to the skin of animals. Most drugs are present in solution in both ionized and nonionized forms. However, usually only lipid soluble or lipophilic drugs readily cross cell membranes. It has been discovered that even non-lipophilic drugs can cross cell membranes if the membrane to be crossed is treated with a penetration enhancer. In addition to aiding the diffusion of non-lipophilic drugs across cell membranes, penetration enhancers also enhance the permeability of lipophilic drugs.


Penetration enhancers can be classified as belonging to one of five broad categories, i.e., surfactants, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants (see e.g., Malmsten, M. Surfactants and polymers in drug delivery, Informa Health Care, New York, NY, 2002; Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p. 92). Each of the above mentioned classes of penetration enhancers and their use in manufacture of pharmaceutical compositions and delivery of pharmaceutical agents are well known in the art.


v. Excipients


In contrast to a carrier compound, a “pharmaceutical carrier” or “excipient” is a pharmaceutically acceptable solvent, suspending agent, or any other pharmacologically inert vehicle for delivering one or more nucleic acids to an animal. The excipient can be liquid or solid and is selected, with the planned manner of administration in mind, so as to provide for the desired bulk, consistency, etc., when combined with a nucleic acid and the other components of a given pharmaceutical composition. Such agent are well known in the art.


vi. Other Components


The compositions of the present invention can additionally contain other adjunct components conventionally found in pharmaceutical compositions, at their art-established usage levels. Thus, for example, the compositions can contain additional, compatible, pharmaceutically-active materials such as, for example, antipruritics, astringents, local anesthetics or anti-inflammatory agents, or can contain additional materials useful in physically formulating various dosage forms of the compositions of the present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers. However, such materials, when added, should not unduly interfere with the biological activities of the components of the compositions of the present invention. The formulations can be sterilized and, if desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings, or aromatic substances, and the like which do not deleteriously interact with the nucleic acid(s) of the formulation.


Aqueous suspensions can contain substances which increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol, or dextran. The suspension can also contain stabilizers.


In some embodiments, pharmaceutical compositions featured in the invention include (a) one or more iRNA and (b) one or more agents which function by a non-iRNA mechanism and which are useful in treating a complement component C3-associated disorder, e.g., hemolysis.


Toxicity and prophylactic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose prophylactically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50/ED50. Compounds that exhibit high therapeutic indices are preferred.


The data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of compositions featured herein in the invention lies generally within a range of circulating concentrations that include the ED50, preferably an ED80 or ED90, with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the methods featured in the invention, the prophylactically effective dose can be estimated initially from cell culture assays. A dose can be formulated in animal models to achieve a circulating plasma concentration range of the compound or, when appropriate, of the polypeptide product of a target sequence (e.g., achieving a decreased concentration of the polypeptide) that includes the IC50 (i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) or higher levels of inhibition as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma can be measured, for example, by high performance liquid chromatography.


In addition to their administration, as discussed above, the iRNAs featured in the invention can be administered in combination with other known agents used for the prevention or treatment of a complement component C3-associated disorder, e.g., hemolysis. In any event, the administering physician can adjust the amount and timing of iRNA administration on the basis of results observed using standard measures of efficacy known in the art or described herein.


VI. Methods For Inhibiting Complement Component C3 Expression

The present invention also provides methods of inhibiting expression of a C3 gene in a cell. The methods include contacting a cell with an RNAi agent, e.g., double stranded RNA agent, in an amount effective to inhibit expression of complement component C3 in the cell, thereby inhibiting expression of complement component C3 in the cell.


Contacting of a cell with an iRNA, e.g., a double stranded RNA agent, may be done in vitro or in vivo. Contacting a cell in vivo with the iRNA includes contacting a cell or group of cells within a subject, e.g., a human subject, with the iRNA. Combinations of in vitro and in vivo methods of contacting a cell are also possible. Contacting a cell may be direct or indirect, as discussed above. Furthermore, contacting a cell may be accomplished via a targeting ligand, including any ligand described herein or known in the art. In preferred embodiments, the targeting ligand is a carbohydrate moiety, e.g., a GalNAc3 ligand, or any other ligand that directs the RNAi agent to a site of interest.


The term “inhibiting,” as used herein, is used interchangeably with “reducing,” “silencing,” “downregulating”, “suppressing”, and other similar terms, and includes any level of inhibition.


The phrase “inhibiting expression of a complement component C3” is intended to refer to inhibition of expression of any complement component C3 gene (such as, e.g., a mouse complement component C3 gene, a rat complement component C3 gene, a monkey complement component C3 gene, or a human complement component C3 gene) as well as variants or mutants of a complement component C3 gene. Thus, the complement component C3 gene may be a wild-type complement component C3 gene, a mutant complement component C3 gene, or a transgenic complement component C3 gene in the context of a genetically manipulated cell, group of cells, or organism.


“Inhibiting expression of a complement component C3 gene” includes any level of inhibition of a complement component C3 gene, e.g., at least partial suppression of the expression of a complement component C3 gene. The expression of the complement component C3 gene may be assessed based on the level, or the change in the level, of any variable associated with complement component C3 gene expression, e.g., complement component C3 mRNA level or complement component C3 protein level. This level may be assessed in an individual cell or in a group of cells, including, for example, a sample derived from a subject. It is understood that complement component C3 is expressed predominantly in the liver, but also in the brain, gall bladder, heart, and kidney, and is present in circulation.


Inhibition may be assessed by a decrease in an absolute or relative level of one or more variables that are associated with complement component C3 expression compared with a control level. The control level may be any type of control level that is utilized in the art, e.g., a pre-dose baseline level, or a level determined from a similar subject, cell, or sample that is untreated or treated with a control (such as, e.g., buffer only control or inactive agent control).


In some embodiments of the methods of the invention, expression of a complement component C3 gene is inhibited by at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or to below the level of detection of the assay. In preferred embodiments, expression of a complement component C3 gene is inhibited by at least 70%. It is further understood that inhibition of complement component C3 expression in certain tissues, e.g., in liver, without a significant inhibition of expression in other tissues, e.g., brain, may be desirable. In preferred embodiments, expression level is determined using the assay method provided in Example 2 with a 10 nM siRNA concentration in the appropriate species matched cell line.


In certain embodiments, inhibition of expression in vivo is determined by knockdown of the human gene in a rodent expressing the human gene, e.g., an AAV-infected mouse expressing the human target gene (i.e., complement component C3), e.g., when administered as a single dose, e.g., at 3 mg/kg at the nadir of RNA expression. Knockdown of expression of an endogenous gene in a model animal system can also be determined, e.g., after administration of a single dose at, e.g., 3 mg/kg at the nadir of RNA expression. Such systems are useful when the nucleic acid sequence of the human gene and the model animal gene are sufficiently close such that the human iRNA provides effective knockdown of the model animal gene. RNA expression in liver is determined using the PCR methods provided in Example 2.


Inhibition of the expression of a complement component C3 gene may be manifested by a reduction of the amount of mRNA expressed by a first cell or group of cells (such cells may be present, for example, in a sample derived from a subject) in which a complement component C3 gene is transcribed and which has or have been treated (e.g., by contacting the cell or cells with an iRNA of the invention, or by administering an iRNA of the invention to a subject in which the cells are or were present) such that the expression of a complement component C3 gene is inhibited, as compared to a second cell or group of cells substantially identical to the first cell or group of cells but which has not or have not been so treated (control cell(s) not treated with an iRNA or not treated with an iRNA targeted to the gene of interest). In preferred embodiments, the inhibition is assessed by the method provided in Example 2 using a 10 nM siRNA concentration in the species matched cell line and expressing the level of mRNA in treated cells as a percentage of the level of mRNA in control cells, using the following formula:










(

mRNA


in


control


cells

)

-

(

mRNA


in


treated


cells

)



(

mRNA


in


control


cells

)


·
100


%




In other embodiments, inhibition of the expression of a complement component C3 gene may be assessed in terms of a reduction of a parameter that is functionally linked to complement component C3 gene expression, e.g., complement component C3 protein level in blood or serum from a subject. Complement component C3 gene silencing may be determined in any cell expressing complement component C3, either endogenous or heterologous from an expression construct, and by any assay known in the art.


Inhibition of the expression of a complement component C3 protein may be manifested by a reduction in the level of the complement component C3 protein that is expressed by a cell or group of cells or in a subject sample (e.g., the level of protein in a blood sample derived from a subject). As explained above, for the assessment of mRNA suppression, the inhibition of protein expression levels in a treated cell or group of cells may similarly be expressed as a percentage of the level of protein in a control cell or group of cells, or the change in the level of protein in a subject sample, e.g., blood or serum derived therefrom.


A control cell, a group of cells, or subject sample that may be used to assess the inhibition of the expression of a complement component C3 gene includes a cell, group of cells, or subject sample that has not yet been contacted with an RNAi agent of the invention. For example, the control cell, group of cells, or subject sample may be derived from an individual subject (e.g., a human or animal subject) prior to treatment of the subject with an RNAi agent or an appropriately matched population control.


The level of complement component C3 mRNA that is expressed by a cell or group of cells may be determined using any method known in the art for assessing mRNA expression. In one embodiment, the level of expression of complement component C3 in a sample is determined by detecting a transcribed polynucleotide, or portion thereof, e.g., mRNA of the complement component C3 gene. RNA may be extracted from cells using RNA extraction techniques including, for example, using acid phenol/guanidine isothiocyanate extraction (RNAzol B; Biogenesis), RNeasy™ RNA preparation kits (Qiagen®) or PAXgene™ (PreAnalytix™, Switzerland). Typical assay formats utilizing ribonucleic acid hybridization include nuclear run-on assays, RT-PCR, RNase protection assays, northern blotting, in situ hybridization, and microarray analysis.


In some embodiments, the level of expression of complement component C3 is determined using a nucleic acid probe. The term “probe”, as used herein, refers to any molecule that is capable of selectively binding to a specific complement component C3. Probes can be synthesized by one of skill in the art, or derived from appropriate biological preparations. Probes may be specifically designed to be labeled. Examples of molecules that can be utilized as probes include, but are not limited to, RNA, DNA, proteins, antibodies, and organic molecules.


Isolated mRNA can be used in hybridization or amplification assays that include, but are not limited to, Southern or northern analyses, polymerase chain reaction (PCR) analyses and probe arrays. One method for the determination of mRNA levels involves contacting the isolated mRNA with a nucleic acid molecule (probe) that can hybridize to complement component C3 mRNA. In one embodiment, the mRNA is immobilized on a solid surface and contacted with a probe, for example by running the isolated mRNA on an agarose gel and transferring the mRNA from the gel to a membrane, such as nitrocellulose. In an alternative embodiment, the probe(s) are immobilized on a solid surface and the mRNA is contacted with the probe(s), for example, in an Affymetrix® gene chip array. A skilled artisan can readily adapt known mRNA detection methods for use in determining the level of complement component C3 mRNA.


An alternative method for determining the level of expression of complement component C3 in a sample involves the process of nucleic acid amplification or reverse transcriptase (to prepare cDNA) of for example mRNA in the sample, e.g., by RT-PCR (the experimental embodiment set forth in Mullis, 1987, U.S. Pat. No. 4,683,202), ligase chain reaction (Barany (1991) Proc. Natl. Acad. Sci. USA 88:189-193), self sustained sequence replication (Guatelli et al. (1990) Proc. Natl. Acad. Sci. USA 87:1874-1878), transcriptional amplification system (Kwoh et al. (1989) Proc. Natl. Acad. Sci. USA 86:1173-1177), Q-Beta Replicase (Lizardi et al. (1988) Bio/Technology 6:1197), rolling circle replication (Lizardi et al., U.S. Pat. No. 5,854,033) or any other nucleic acid amplification method, followed by the detection of the amplified molecules using techniques well known to those of skill in the art. These detection schemes are especially useful for the detection of nucleic acid molecules if such molecules are present in very low numbers. In particular aspects of the invention, the level of expression of C3 is determined by quantitative fluorogenic RT-PCR (i.e., the TaqMan™ System). In preferred embodiments, expression level is determined by the method provided in Example 2 using, e.g., a 10 nM siRNA concentration, in the species matched cell line.


The expression levels of complement component C3 mRNA may be monitored using a membrane blot (such as used in hybridization analysis such as northern, Southern, dot, and the like), or microwells, sample tubes, gels, beads or fibers (or any solid support comprising bound nucleic acids). See U.S. Pat. Nos. 5,770,722, 5,874,219, 5,744,305, 5,677,195 and 5,445,934, which are incorporated herein by reference. The determination of complement component C3 expression level may also comprise using nucleic acid probes in solution.


In preferred embodiments, the level of mRNA expression is assessed using branched DNA (bDNA) assays or real time PCR (qPCR). The use of these methods is described and exemplified in the Examples presented herein. In preferred embodiments, expression level is determined by the method provided in Example 2 using a 10 nM siRNA concentration in the species matched cell line.


The level of C3 protein expression may be determined using any method known in the art for the measurement of protein levels. Such methods include, for example, electrophoresis, capillary electrophoresis, high performance liquid chromatography (HPLC), thin layer chromatography (TLC), hyperdiffusion chromatography, fluid or gel precipitin reactions, absorption spectroscopy, a colorimetric assays, spectrophotometric assays, flow cytometry, immunodiffusion (single or double), immunoelectrophoresis, western blotting, radioimmunoassay (RIA), enzyme-linked immunosorbent assays (ELISAs), immunofluorescent assays, electrochemiluminescence assays, and the like.


In some embodiments, the efficacy of the methods of the invention are assessed by a decrease in C3 mRNA or protein level (e.g., in a liver biopsy).


In some embodiments of the methods of the invention, the iRNA is administered to a subject such that the iRNA is delivered to a specific site within the subject. The inhibition of expression of complement component C3 may be assessed using measurements of the level or change in the level of complement component C3 mRNA or complement component C3 protein in a sample derived from fluid or tissue from the specific site within the subject (e.g., liver or blood).


As used herein, the terms detecting or determining a level of an analyte are understood to mean performing the steps to determine if a material, e.g., protein, RNA, is present. As used herein, methods of detecting or determining include detection or determination of an analyte level that is below the level of detection for the method used.


VII. Prophylactic and Treatment Methods of the Invention

The present invention also provides methods of using an iRNA of the invention or a composition containing an iRNA of the invention to inhibit expression of complement component C3, thereby preventing or treating a complement component C3-associated disorder, e.g., cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy.


In the methods of the invention the cell may be contacted with the siRNA in vitro or in vivo, i.e., the cell may be within a subject.


A cell suitable for treatment using the methods of the invention may be any cell that expresses a complement component C3 gene, e.g., a liver cell, a brain cell, a gall bladder cell, a heart cell, or a kidney cell, but preferably a liver cell. A cell suitable for use in the methods of the invention may be a mammalian cell, e.g., a primate cell (such as a human cell, including human cell in a chimeric non-human animal, or a non-human primate cell, e.g., a monkey cell or a chimpanzee cell), or a non-primate cell. In certain embodiments, the cell is a human cell, e.g., a human liver cell. In the methods of the invention, complement component C3 expression is inhibited in the cell by at least 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95, or to a level below the level of detection of the assay.


The in vivo methods of the invention may include administering to a subject a composition containing an iRNA, where the iRNA includes a nucleotide sequence that is complementary to at least a part of an RNA transcript of the complement component C3 gene of the mammal to which the RNAi agent is to be administered. The composition can be administered by any means known in the art including, but not limited to oral, intraperitoneal, or parenteral routes, including intracranial (e.g., intraventricular, intraparenchymal, and intrathecal), intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), nasal, rectal, and topical (including buccal and sublingual) administration. In certain embodiments, the compositions are administered by intravenous infusion or injection. In certain embodiments, the compositions are administered by subcutaneous injection. In certain embodiments, the compositions are administered by intramuscular injection.


In one aspect, the present invention also provides methods for inhibiting the expression of a complement component C3 gene in a mammal. The methods include administering to the mammal a composition comprising a dsRNA that targets a complement component C3 gene in a cell of the mammal and maintaining the mammal for a time sufficient to obtain degradation of the mRNA transcript of the complement component C3 gene, thereby inhibiting expression of the complement component C3 gene in the cell. Reduction in gene expression can be assessed by any methods known in the art and by methods, e.g. qRT-PCR, described herein, e.g., in Example 2. Reduction in protein production can be assessed by any methods known it the art, e.g. ELISA. In certain embodiments, a puncture liver biopsy sample serves as the tissue material for monitoring the reduction in the complement component C3 gene or protein expression. In other embodiments, a blood sample serves as the subject sample for monitoring the reduction in the complement component C3 protein expression.


The present invention further provides methods of treatment in a subject in need thereof, e.g., a subject diagnosed with a complement component C3-associated disorder, such as, cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), or C3 glomerulopathy.


The present invention further provides methods of prophylaxis in a subject in need thereof. The treatment methods of the invention include administering an iRNA of the invention to a subject, e.g., a subject that would benefit from a reduction of complement component C3 expression, in a prophylactically effective amount of an iRNA targeting a complement component C3 gene or a pharmaceutical composition comprising an iRNA targeting a complement component C3 gene.


In one embodiment, a complement component C3-associated disease is selected from the group consisting of cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy.


In one embodiment, a complement component C3-associated disease is cold agglutinin disease (CAD). CAD is an autoimmune complement component C3-induced hemolytic anemia in which cold exposure causes clinical symptoms related to agglutination of red blood cells (RBCs) in cold parts of the body (e.g., livedo reticularis or acrocyanosis) and hemolytic anemia. Cold agglutinins are IgM antibodies that recognize antigens on red blood cells (RBCs) at temperatures below normal core body temperature. They can cause agglutination of the RBCs, complement activation and extravascular hemolysis, resulting in anemia, typically without hemoglobinuria. The CAD may be primary CAD (also called idiopathic CAD) or secondary CAD. In subjects having primary CAD, cold agglutinins cause RBC agglutination and extravascular hemolysis in the absence of an underlying disorder. In subjects having secondary CAD (also referred to as cold agglutinin syndrome, or CAS), cold agglutinins arise in the setting of an underlying disorder such as a viral infection, autoimmune disorder, or lymphoid malignancy (see, e.g., Berentsen (2015) Transfus Med Hemother 42:303-310).


In one embodiment, a complement component C3-associated disease is warm autoimmune hemolytic anemia. Warm autoimmune hemolytic anemia is an autoimmune complement component C3-induced hemolytic anemia in which red blood cells (RBCs) agglutinate in parts of the body at temperatures equal to or greater than normal body temperature and hemolytic anemia as a result of IgG antibodies directed against blood group antigens which activate the complement system. Warm autoimmune hemolytic anemia is the most common type of autoimmune hemolytic anemia, comprising ˜70% to 80% of all adult cases and ˜50% of the pediatric cases. About half of the warm autoimmune hemolytic anemia cases are primary because no specific etiology can be found, whereas the rest are recognized as secondary to lymphoproliferative syndromes; malignant diseases including chronic lymphoblastic leukemia (CLL), non-Hodgkin's lymphoma, and solid tumors; rheumatologic diseases, especially systemic lupus erythematosus; infections (mostly viral); drugs; frequent cephalosporins and piperacillin; or a previous transfusion or transplantation (see, e.g., Berentsen (2015) Transfus Med Hemother 42:303-310).


In one embodiment, a complement component C3-associated disease is paroxysmal nocturnal hemoglobinuria (PNH). The PNH may be classical PNH or PNH in the setting of another bone marrow failure syndrome and/or myelodysplastic syndromes (MDS), e.g., cytopenias. PNH is an acquired autoimmune disorder that leads to the premature death and impaired production of blood cells, characterized by complement-mediated hemolytic anemia, thrombophilia, and bone marrow failure (see, e.g., Risitano (2013) Adv Exp Med Biol 735:155).


In one embodiment, a complement component C3-associated disease is lupis nephritis (LN), i.e., any one of Class I-Class VI lupus nephritis). LN is a type of glomerulonephritis caused by systemic lupus erythematosus (SLE). Lupus nephritis occurs due to immune complex deposition in any or all renal compartments, including the glomeruli, tubules, and interstitium. IgG is the most prevalent antibody found but IgM, and IgA can be seen as well. These auto-antibodies cause activation of both the classic and alternative complement pathways and so C1, C3 and properdin may be found on biopsy.


In one embodiment, a complement component C3-associated disease is bullous pemphigoid. Bullous pemphigoid an autoimmune blistering disease induced by autoantibodies against type XVII collagen (COL17) that activates complement and subsequently recruits inflammatory cells at the dermal/epidermal junction. Bullous pemphigoid is the most common autoimmune blistering disorder characterized by tense blisters with itchy urticarial erythema and plaques that develop on the entire body.


In one embodiment, a complement component C3-associated disease is pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF). Pemphigus is a group of rare chronic blistering diseases characterized by IgG-autoantibodies directed against a variety of desmosomal transmembrane glycoproteins and intracellular deposition of IgG and C3c. Patients with pemphigus vulgaris typically present with lesions of the oral mucosa followed by skin-involvement and autoantibodies are directed against epithelial adhesion protein desmoglein 3 and/or desmoglein 1. In pemphigus foliaceus the lesions are localized on the skin, without involvement of the mucous membranes, and autoantibodies are directed against desmoglein 1. In one embodiment, the pemphigus is pemphigus vulgaris (PV). In another embodiment, the pemphigus is pemphigus foliaceus (PF).


In one embodiment, a complement component C3-associated disease is C3 glomerulopathy. C3 glomerulopathy is characterized by activation of the alternative complement cascade and deposition of complement component C3 without any immunoglobulin deposits in the glomeruli of the kidney.


An iRNA of the invention may be administered as a “free iRNA.” A free iRNA is administered in the absence of a pharmaceutical composition. The naked iRNA may be in a suitable buffer solution. The buffer solution may comprise acetate, citrate, prolamine, carbonate, or phosphate, or any combination thereof. In one embodiment, the buffer solution is phosphate buffered saline (PBS). The pH and osmolarity of the buffer solution containing the iRNA can be adjusted such that it is suitable for administering to a subject.


Alternatively, an iRNA of the invention may be administered as a pharmaceutical composition, such as a dsRNA liposomal formulation.


Subjects that would benefit from an inhibition of complement component C3 gene expression are subjects susceptible to or diagnosed with a complement component C3-associated disorder, such as cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy.


In an embodiment, the method includes administering a composition featured herein such that expression of the target complement component C3 gene is decreased, such as for about 1, 2, 3, 4, 5, 6, 1-6, 1-3, or 3-6 months per dose. In certain embodiments, the composition is administered once every 3-6 months.


Preferably, the iRNAs useful for the methods and compositions featured herein specifically target RNAs (primary or processed) of the target complement component C3 gene. Compositions and methods for inhibiting the expression of these genes using iRNAs can be prepared and performed as described herein.


Administration of the iRNA according to the methods of the invention may result prevention or treatment of a complement component C3-associated disorder, e.g., cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, pemphigus, e.g., pemphigus vulgaris (PV) and pemphigus foliaceus (PF), and C3 glomerulopathy.


Subjects can be administered a therapeutic amount of iRNA, such as about 0.01 mg/kg to about 200 mg/kg.


The iRNA is preferably administered subcutaneously, i.e., by subcutaneous injection. One or more injections may be used to deliver the desired dose of iRNA to a subject. The injections may be repeated over a period of time.


The administration may be repeated on a regular basis. In certain embodiments, after an initial treatment regimen, the treatments can be administered on a less frequent basis. A repeat-dose regimen may include administration of a therapeutic amount of iRNA on a regular basis, such as once per month to once a year. In certain embodiments, the iRNA is administered about once per month to about once every three months, or about once every three months to about once every six months.


The invention further provides methods and uses of an iRNA agent or a pharmaceutical composition thereof for treating a subject that would benefit from reduction and/or inhibition of C3 gene expression, e.g., a subject having a C3-associated disease, in combination with other pharmaceuticals and/or other therapeutic methods, e.g., with known pharmaceuticals and/or known therapeutic methods, such as, for example, those which are currently employed for treating these disorders.


Accordingly, in some aspects of the invention, the methods which include either a single iRNA agent of the invention, further include administering to the subject one or more additional therapeutic agents.


The iRNA agent and an additional therapeutic agent and/or treatment may be administered at the same time and/or in the same combination, e.g., parenterally, or the additional therapeutic agent can be administered as part of a separate composition or at separate times and/or by another method known in the art or described herein.


For example, additional therapeutics and therapeutic methods suitable for treating a subject that would benefit from reducton in C3 expression, e.g., a subject having a complement component C3-associated disease, include plasmaphoresis, thrombolytic therapy (e.g., streptokinase), antiplatelet agents, folic acid, corticosteroids; immunosuppressive agents; estrogens, methotrexate, 6-MP, azathioprine sulphasalazine, mesalazine, olsalazine, chloroquinine/hydroxychloroquine, pencillamine, aurothiomalate (intramuscular and oral), azathioprine, cochicine, corticosteroids (oral, inhaled and local injection), beta-2 adrenoreceptor agonists (salbutamol, terbutaline, salmeteral), xanthines (theophylline, aminophylline), cromoglycate, nedocromil, ketotifen, ipratropium and oxitropium, cyclosporin, FK506, rapamycin, mycophenolate mofetil, leflunomide, NSAIDs, for example, ibuprofen, corticosteroids such as prednisolone, phosphodiesterase inhibitors, adensosine agonists, antithrombotic agents, complement inhibitors, adrenergic agents, agents which interfere with signalling by proinflammatory cytokines, such as TNF-α or IL-1 (e.g., IRAK, NIK, IKK, p38 or MAP kinase inhibitors), IL-1β converting enzyme inhibitors, TNFαconverting enzyme (TACE) inhibitors, T-cell signalling inhibitors, such as kinase inhibitors, metalloproteinase inhibitors, sulfasalazine, azathioprine, 6-mercaptopurines, angiotensin converting enzyme inhibitors, soluble cytokine receptors and derivatives thereof (e.g., soluble p55 or p75 TNF receptors and the derivatives p75TNFRIgG (Enbrel™ and p55TNFRIgG (Lenercept)), sIL-1RI, sIL-1RII, and sIL-6R), antiinflammatory cytokines (e.g., IL-4, IL-10, IL-11, IL-13 and TGFβ), celecoxib, folic acid, hydroxychloroquine sulfate, rofecoxib, etanercept, infliximonoclonal antibody, naproxen, valdecoxib, sulfasalazine, methylprednisolone, meloxicam, methylprednisolone acetate, gold sodium thiomalate, aspirin, triamcinolone acetonide, propoxyphene napsylate/apap, folate, nabumetone, diclofenac, piroxicam, etodolac, diclofenac sodium, oxaprozin, oxycodone hydrochloride, hydrocodone bitartrate/apap, diclofenac sodium/misoprostol, fentanyl, anakinra, human recombinant, tramadol hydrochloride, salsalate, sulindac, cyanocobalamin/folic acid/pyridoxine, acetaminophen, alendronate sodium, prednisolone, morphine sulfate, lidocaine hydrochloride, indomethacin, glucosamine sulf/chondroitin, amitriptyline hydrochloride, sulfadiazine, oxycodone hydrochloride/acetaminophen, olopatadine hydrochloride, misoprostol, naproxen sodium, omeprazole, cyclophosphamide, rituximonoclonal antibody, IL-1 TRAP, MRA, CTLA4-IG, IL-18 BP, anti-IL-18, Anti-IL15, BIRB-796, SCIO-469, VX-702, AMG-548, VX-740, Roflumilast, IC-485, CDC-801, Mesopram, cyclosporine, cytokine suppressive anti-inflammatory drug(s) (CSAIDs); CDP-571/BAY-10-3356 (humanized anti-TNFα antibody; Celltech/Bayer); cA2/infliximonoclonal antibody (chimeric anti-TNFα antibody; Centocor); 75 kdTNFR-IgG/etanercept (75 kD TNF receptor-IgG fusion protein; Immunex; see e.g., (1994) Arthr. Rheum. 37: S295; (1996) J. Invest. Med. 44: 235A); 55 kdTNF-IgG (55 kD TNF receptor-IgG fusion protein; Hoffmann-LaRoche); IDEC-CE9.1/SB 210396 (non-depleting primatized anti-CD4 antibody; IDEC/SmithKline; see e.g., (1995) Arthr. Rheum. 38: S185); DAB 486-IL-2 and/or DAB 389-IL-2 (IL-2 fusion proteins; Seragen; see e.g., (1993) Arthrit. Rheum. 36: 1223); Anti-Tac (humanized anti-IL-2Rα; Protein Design Labs/Roche); IL-4 (anti-inflammatory cytokine; DNAX/Schering); IL-10 (SCH 52000; recombinant IL-10, anti-inflammatory cytokine; DNAX/Schering); IL-4; IL-10 and/or IL-4 agonists (e.g., agonist antibodies); IL-IRA (IL-1 receptor antagonist; Synergen/Amgen); anakinra (Kineret®/Amgen); TNF-bp/s-TNF (soluble TNF binding protein; see e.g., (1996) Arthr. Rheum. 39(9 (supplement)): S284; (1995) Amer. J. Physiol. —Heart and Circ. Physiol. 268: 37-42); R973401 (phosphodiesterase Type IV inhibitor; see e.g., (1996) Arthr. Rheum. 39(9 (supplement): 5282); MK-966 (COX-2 Inhibitor; see e.g., (1996) Arthr. Rheum. 39(9 (supplement): S81); Iloprost (see e.g., (1996) Arthr. Rheum. 39(9 (supplement): S82); methotrexate; thalidomide (see e.g., (1996) Arthr. Rheum. 39(9 (supplement): 5282) and thalidomide-related drugs (e.g., Celgen); leflunomide (anti-inflammatory and cytokine inhibitor; see e.g., (1996) Arthr. Rheum. 39(9 (supplement): S131; (1996) Inflamm. Res. 45: 103-107); tranexamic acid (inhibitor of plasminogen activation; see e.g., (1996) Arthr. Rheum. 39(9 (supplement): S284); T-614 (cytokine inhibitor; see e.g., (1996) Arthr. Rheum. 39(9 (supplement): S282); prostaglandin E1 (see e.g., (1996) Arthr. Rheum. 39(9 (supplement): S282); Tenidap (non-steroidal anti-inflammatory drug; see e.g., (1996) Arthr. Rheum. 39(9 (supplement): S280); Naproxen (non-steroidal anti-inflammatory drug; see e.g., (1996) Neuro. Report 7: 1209-1213); Meloxicam (non-steroidal anti-inflammatory drug); Ibuprofen (non-steroidal anti-inflammatory drug); Piroxicam (non-steroidal anti-inflammatory drug); Diclofenac (non-steroidal anti-inflammatory drug); Indomethacin (non-steroidal anti-inflammatory drug); Sulfasalazine (see e.g., (1996) Arthr. Rheum. 39(9 (supplement): S281); Azathioprine (see e.g., (1996) Arthr. Rheum. 39(9 (supplement): 5281); ICE inhibitor (inhibitor of the enzyme interleukin-1 □ converting enzyme); zap-70 and/or lck inhibitor (inhibitor of the tyrosine kinase zap-70 or lck); VEGF inhibitor and/or VEGF-R inhibitor (inhibitors of vascular endothelial cell growth factor or vascular endothelial cell growth factor receptor; inhibitors of angiogenesis); corticosteroid anti-inflammatory drugs (e.g., SB203580); TNF-convertase inhibitors; anti-IL-12 antibodies; anti-IL-18 antibodies; interleukin-11 (see e.g., (1996) Arthr. Rheum. 39(9 (supplement): S296); interleukin-13 (see e.g., (1996) Arthr. Rheum. 39(9 (supplement): S308); interleukin-17 inhibitors (see e.g., (1996) Arthr. Rheum. 39(9 (supplement): S120); gold; penicillamine; chloroquine; chlorambucil; hydroxychloroquine; cyclosporine; cyclophosphamide; total lymphoid irradiation; anti-thymocyte globulin; anti-CD4 antibodies; CD5-toxins; orally-administered peptides and collagen; lobenzarit disodium; Cytokine Regulating Agents (CRAs) HP228 and HP466 (Houghten Pharmaceuticals, Inc.); ICAM-1 antisense phosphorothioate oligo-deoxynucleotides (ISIS 2302; Isis Pharmaceuticals, Inc.); soluble complement receptor 1 (TP10; T Cell Sciences, Inc.); prednisone; orgotein; glycosaminoglycan polysulphate; minocycline; anti-IL2R antibodies; marine and botanical lipids (fish and plant seed fatty acids; see e.g., DeLuca et al. (1995) Rheum. Dis. Clin. North Am. 21: 759-777); auranofin; phenylbutazone; meclofenamic acid; flufenamic acid; intravenous immune globulin; zileuton; azaribine; mycophenolic acid (RS-61443); tacrolimus (FK-506); sirolimus (rapamycin); amiprilose (therafectin); cladribine (2-chlorodeoxyadenosine); methotrexate; bcl-2 inhibitors (see Bruncko, M. et al. (2007) J. Med. Chem. 50(4): 641-662); antivirals and immune-modulating agents, small molecule inhibitor of KDR, small molecule inhibitor of Tie-2; methotrexate; prednisone; celecoxib; folic acid; hydroxychloroquine sulfate; rofecoxib; etanercept; infliximonoclonal antibody; leflunomide; naproxen; valdecoxib; sulfasalazine; methylprednisolone; ibuprofen; meloxicam; methylprednisolone acetate; gold sodium thiomalate; aspirin; azathioprine; triamcinolone acetonide; propxyphene napsylate/apap; folate; nabumetone; diclofenac; piroxicam; etodolac; diclofenac sodium; oxaprozin; oxycodone hcl; hydrocodone bitartrate/apap; diclofenac sodium/misoprostol; fentanyl; anakinra, human recombinant; tramadol hcl; salsalate; sulindac; cyanocobalamin/fa/pyridoxine; acetaminophen; alendronate sodium; prednisolone; morphine sulfate; lidocaine hydrochloride; indomethacin; glucosamine sulfate/chondroitin; cyclosporine; amitriptyline hydrochloride; sulfadiazine; oxycodone hcl/acetaminophen; olopatadine hcl; misoprostol; naproxen sodium; omeprazole; mycophenolate mofetil; cyclophosphamide; rituximonoclonal antibody; IL-1 TRAP; MRA; CTLA4-IG; IL-18 BP; IL-12/23; anti-IL 18; anti-IL 15; BIRB-796; SCIO-469; VX-702; AMG-548; VX-740; Roflumilast; IC-485; CDC-801; mesopram, albuterol, salmeterol/fluticasone, montelukast sodium, fluticasone propionate, budesonide, prednisone, salmeterol xinafoate, levalbuterol hcl, albuterol sulfate/ipratropium, prednisolone sodium phosphate, triamcinolone acetonide, beclomethasone dipropionate, ipratropium bromide, azithromycin, pirbuterol acetate, prednisolone, theophylline anhydrous, methylprednisolone sodium succinate, clarithromycin, zafirlukast, formoterol fumarate, influenza virus vaccine, methylprednisolone, amoxicillin trihydrate, flunisolide, allergy injection, cromolyn sodium, fexofenadine hydrochloride, flunisolide/menthol, amoxicillin/clavulanate, levofloxacin, inhaler assist device, guaifenesin, dexamethasone sodium phosphate, moxifloxacin hcl, doxycycline hyclate, guaifenesin/d-methorphan, p-ephedrine/cod/chlorphenir, gatifloxacin, cetirizine hydrochloride, mometasone furoate, salmeterol xinafoate, benzonatate, cephalexin, pe/hydrocodone/chlorphenir, cetirizine hcl/pseudoephed, phenylephrine/cod/promethazine, codeine/promethazine, cefprozil, dexamethasone, guaifenesin/pseudoephedrine, chlorpheniramine/hydrocodone, nedocromil sodium, terbutaline sulfate, epinephrine, methylprednisolone, metaproterenol sulfate, aspirin, nitroglycerin, metoprolol tartrate, enoxaparin sodium, heparin sodium, clopidogrel bisulfate, carvedilol, atenolol, morphine sulfate, metoprolol succinate, warfarin sodium, lisinopril, isosorbide mononitrate, digoxin, furosemide, simvastatin, ramipril, tenecteplase, enalapril maleate, torsemide, retavase, losartan potassium, quinapril hcl/mag carb, bumetanide, alteplase, enalaprilat, amiodarone hydrochloride, tirofiban hcl m-hydrate, diltiazem hydrochloride, captopril, irbesartan, valsartan, propranolol hydrochloride, fosinopril sodium, lidocaine hydrochloride, eptifibatide, cefazolin sodium, atropine sulfate, aminocaproic acid, spironolactone, interferon, sotalol hydrochloride, potassium chloride, docusate sodium, dobutamine hcl, alprazolam, pravastatin sodium, atorvastatin calcium, midazolam hydrochloride, meperidine hydrochloride, isosorbide dinitrate, epinephrine, dopamine hydrochloride, bivalirudin, rosuvastatin, ezetimibe/simvastatin, avasimibe, and cariporide.


In some aspects, the additional therapeutic agent is an iRNA agent targeting a C5 gene, such as described in U.S. Pat. No. 9,249,415, U.S. Provisional Patent Application Nos. 62/174,933, filed on Jun. 12, 2015, 62/263,066, filed on Dec. 4, 2015, the entire contents of each of which are hereby incorporated herein by reference.


In other aspects, the additional therapeutic agent is an anti-complement component C5 antibody, or antigen-binding fragment thereof (e.g., eculizumab). Eculizumab is a humanized monoclonal IgG2/4, kappa light chain antibody that specifically binds complement component C5 with high affinity and inhibits cleavage of C5 to C5a and C5b, thereby inhibiting the generation of the terminal complement complex C5b-9. Eculizumab is described in U.S. Pat. No. 6,355,245, the entire contents of which are incorporated herein by reference.


In yet other aspects, the additional therapeutic is a C3 peptide inhibitor, or analog thereof. In one embodiment, the C3 peptide inhibitor is compstatin. Compstatin is a cyclic tridecapeptide with potent and selective C3 inhibitory activity. Compstatin, and its analogs, are described in U.S. Pat. Nos. 7,888,323, 7,989,589, and 8,442,776, in U.S. Patent Publication No. 2012/0178694 and 2013/0053302, and in PCT Publication Nos. WO 2012/174055, WO 2012/2178083, WO 2013/036778, the entire contents of each of which are incorporated herein by reference.


VIII. Kits

The present invention also provides kits for performing any of the methods of the invention. Such kits include one or more dsRNA agent(s) and instructions for use, e.g., instructions for administering a prophylactically or therapeutically effective amount of a dsRNA agent(s). The dsRNA agent may be in a vial or a pre-filled syringe. The kits may optionally further comprise means for administering the dsRNA agent (e.g., an injection device, such as a pre-filled syringe), or means for measuring the inhibition of C3 (e.g., means for measuring the inhibition of C3 mRNA, C3 protein, and/or C3 activity). Such means for measuring the inhibition of C3 may comprise a means for obtaining a sample from a subject, such as, e.g., a plasma sample. The kits of the invention may optionally further comprise means for determining the therapeutically effective or prophylactically effective amount.


This invention is further illustrated by the following examples which should not be construed as limiting. The entire contents of all references, patents and published patent applications cited throughout this application, as well as the informal Sequence Listing and Figures, are hereby incorporated herein by reference.


EXAMPLES
Example 1. iRNA Synthesis
Source of Reagents

Where the source of a reagent is not specifically given herein, such reagent can be obtained from any supplier of reagents for molecular biology at a quality/purity standard for application in molecular biology.


siRNA Design


siRNAs targeting the human complement component C3 (C3) gene (human: NCBI refseqID NM_000064.3; NCBI GeneID: 718) were designed using custom R and Python scripts. The human NM_000064.3 REFSEQ mRNA, has a length of 5148 bases.


Detailed lists of the unmodified complement component sense and antisense strand nucleotide sequences are shown in Tables 2, 4, and 6. Detailed lists of the modified complement component C3 sense and antisense strand nucleotide sequences are shown in Tables 3, 5, and 7.


It is to be understood that, throughout the application, a duplex name without a decimal is equivalent to a duplex name with a decimal which merely references the batch number of the duplex.


For example, AD-564727 is equivalent to AD-564727.1.


siRNA Synthesis


siRNAs were synthesized and annealed using routine methods known in the art.


Briefly, siRNA sequences were synthesized at 1 μmol scale on a Mermade 192 synthesizer (BioAutomation) using the solid support mediated phosphoramidite chemistry. The solid support was controlled pore glass (500 A) loaded with custom GalNAc ligand or universal solid support (AM biochemical). Ancillary synthesis reagents, 2′-F and 2′-O-Methyl RNA and deoxy phosphoramidites were obtained from Thermo-Fisher (Milwaukee, WI) and Hongene (China). 2′F 2′-O-Methyl, GNA (glycol nucleic acids), 5′ phosphate and other modifications were introduced using the corresponding phosphoramidites. Synthesis of 3′ GalNAc conjugated single strands was performed on a GalNAc modified CPG support. Custom CPG universal solid support was used for the synthesis of antisense single strands. Coupling time for all phosphoramidites (100 mM in acetonitrile) was 5 min employing 5-Ethylthio-1H-tetrazole (ETT) as activator (0.6 M in acetonitrile). Phosphorothioate linkages were generated using a 50 mM solution of 3-((Dimethylamino-methylidene)amino)-3H-1,2,4-dithiazole-3-thione (DDTT, obtained from Chemgenes (Wilmington, MA, USA)) in anhydrous acetonitrile/pyridine (1:1 v/v). Oxidation time was 3 minutes. All sequences were synthesized with final removal of the DMT group (“DMT off”).


Upon completion of the solid phase synthesis, oligoribonucleotides were cleaved from the solid support and deprotected in sealed 96 deep well plates using 200 μL Aqueous Methylamine reagents at 60° C. for 20 minutes. For sequences containing 2′ ribo residues (2′-OH) that are protected with a tert-butyl dimethyl silyl (TBDMS) group, a second step deprotection was performed using TEA.3HF (triethylamine trihydro fluoride) reagent. To the methylamine deprotection solution, 200 uL of dimethyl sulfoxide (DMSO) and 300 ul TEA.3HF reagent was added and the solution was incubated for additional 20 min at 60° C. At the end of cleavage and deprotection step, the synthesis plate was allowed to come to room temperature and was precipitated by addition of 1 mL of acetontile: ethanol mixture (9:1). The plates were cooled at −80 C for 2 hrs, superanatant decanted carefully with the aid of a multi channel pipette. The oligonucleotide pellet was re-suspended in 20 mM NaOAc buffer and were desalted using a 5 mL HiTrap size exclusion column (GE Healthcare) on an AKTA Purifier System equipped with an A905 autosampler and a Frac 950 fraction collector. Desalted samples were collected in 96-well plates. Samples from each sequence were analyzed by LC-MS to confirm the identity, UV (260 nm) for quantification and a selected set of samples by IEX chromatography to determine purity.


Annealing of single strands was performed on a Tecan liquid handling robot. Equimolar mixture of sense and antisense single strands were combined and annealed in 96 well plates. After combining the complementary single strands, the 96-well plate was sealed tightly and heated in an oven at 100° C. for 10 minutes and allowed to come slowly to room temperature over a period 2-3 hours. The concentration of each duplex was normalized to 10 M in 1×PBS and then submitted for in vitro screening assays.


Example 2. In Vitro Screening Methods
Cell Culture and 384-Well Transfections

Hep3b cells (ATCC, Manassas, VA) were grown to near confluence at 37° C. in an atmosphere of 5% CO2 in Eagle's Minimum Essential Medium (Gibco) supplemented with 10% FBS (ATCC) before being released from the plate by trypsinization. For mouse cross reactive duplexes, primary mouse hepatocytes (PMH) were freshly isolated less than 1 hour prior to transfections and grown in primary hepatocyte media. For both Hep3B and PMH, transfection was carried out by adding 14.8 μl of Opti-MEM plus 0.2 μl of Lipofectamine RNAiMax per well (Invitrogen, Carlsbad CA. cat #13778-150) to 5 μl of each siRNA duplex to an individual well in a 96-well plate. The mixture was then incubated at room temperature for 15 minutes. Eighty μl of complete growth media without antibiotic containing ˜2×104 Hep3B cells or PMH were then added to the siRNA mixture. Cells were incubated for 24 hours prior to RNA purification. Single dose experiments were performed at 10 nM and 0.1 nM final duplex concentration and dose response experiments were done using 8× 5-fold serial dilutions over the range of 10 nM to 128 pM.


Total RNA Isolation Using DYNABEADS mRNA Isolation Kit (Invitrogen™, Part #: 610-12)


Cells were lysed in 75 μl of Lysis/Binding Buffer containing 3 μL of beads per well and mixed for 10 minutes on an electrostatic shaker. The washing steps were automated on a Biotek EL406, using a magnetic plate support. Beads were washed (in 90 μL) once in Buffer A, once in Buffer B, and twice in Buffer E, with aspiration steps in between. Following a final aspiration, complete 10 μL RT mixture was added to each well, as described below.


cDNA Synthesis Using ABI High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, Cat #4368813)


A master mix of 1 μl 10× Buffer, 0.4 μl 25× dNTPs, 1 μl Random primers, 0.5 μl Reverse Transcriptase, 0.5 μl RNase inhibitor and 6.6 μl of H2O per reaction were added per well. Plates were sealed, agitated for 10 minutes on an electrostatic shaker, and then incubated at 37 degrees C. for 2 hours. Following this, the plates were agitated at 80 degrees C. for 8 minutes.


Real Time PCR

Two microlitre (μl) of cDNA were added to a master mix containing 0.5 μl of human GAPDH TaqMan Probe (4326317E), 0.5 μl human C3, 2 μl nuclease-free water and 5 μl Lightcycler 480 probe master mix (Roche Cat #04887301001) per well in a 384 well plates (Roche cat #04887301001). Real time PCR was done in a LightCycler480 Real Time PCR system (Roche).


To calculate relative fold change, data were analyzed using the ΔΔCt method and normalized to assays performed with cells transfected with 10 nM AD-1955, or mock transfected cells. IC50s were calculated using a 4 parameter fit model using XLFit and normalized to cells transfected with AD-1955 or mock-transfected. The sense and antisense sequences of AD-1955 are: sense: cuuAcGcuGAGuAcuucGAdTsdT (SEQ ID NO: 13) and antisense UCGAAGuACUcAGCGuAAGdTsdT (SEQ ID NO:14).


The results of the screening of the dsRNA agents listed in Tables 2 and 3 in Hep3B cells are shown in Table 8. The results of the screening of the dsRNA agents listed in Tables 2 and 3 in PMH cells are shown in Table 9. The results of the screening of the dsRNA agents listed in Tables 4 and 5 in Hep3B cells are shown in Table 10. The results of the screening of the dsRNA agents listed in Tables 4 and 5 in PMH cells are shown in Table 11. The results of the screening of the dsRNA agents listed in Tables 6 and 7 in Hep3B cells are shown in Table 12. The results of the screening of the dsRNA agents listed in Tables 6 and 7 in PMH cells are shown in Table 13.









TABLE 1







Abbreviations of nucleotide monomers used in nucleic


acid sequence representation. It will be understood


that these monomers, when present in an oligonucleotide,


are mutually linked by 5′-3′-phosphodiester bonds.








Abbreviation
Nucleotide(s)





A
Adenosine-3′-phosphate


Ab
beta-L-adenosine-3′-phosphate


Abs
beta-L-adenosine-3′-phosphorothioate


Af
2′-fluoroadenosine-3′-phosphate


Afs
2′-fluoroadenosine-3′-phosphorothioate


As
adenosine-3′-phosphorothioate


C
cytidine-3′-phosphate


Cb
beta-L-cytidine-3′-phosphate


Cbs
beta-L-cytidine-3′-phosphorothioate


Cf
2′-fluorocytidine-3′-phosphate


Cfs
2′-fluorocytidine-3′-phosphorothioate


Cs
cytidine-3′-phosphorothioate


G
guanosine-3′-phosphate


Gb
beta-L-guanosine-3′-phosphate


Gbs
beta-L-guanosine-3′-phosphorothioate


Gf
2′-fluoroguanosine-3′-phosphate


Gfs
2′-fluoroguanosine-3′-phosphorothioate


Gs
guanosine-3′-phosphorothioate


T
5′-methyluridine-3′-phosphate


Tf
2′-fluoro-5-methyluridine-3′-phosphate


Tfs
2′-fluoro-5-methyluridine-3′-phosphorothioate


Ts
5-methyluridine-3′-phosphorothioate


U
Uridine-3′-phosphate


Uf
2′-fluorouridine-3′-phosphate


Ufs
2′-fluorouridine-3′-phosphorothioate


Us
uridine-3′-phosphorothioate


N
any nucleotide, modified or unmodified


a
2′-O-methyladenosine-3′-phosphate


as
2′-O-methyladenosine-3′-phosphorothioate


c
2′-O-methylcytidine-3′-phosphate


cs
2′-O-methylcytidine-3′-phosphorothioate


g
2′-O-methylguanosine-3′-phosphate


gs
2′-O-methylguanosine-3′-phosphorothioate


t
2′-O-methyl-5-methyluridine-3′-phosphate


ts
2′-O-methyl-5-methyluridine-3′-phosphorothioate


u
2′-O-methyluridine-3′-phosphate


us
2′-O-methyluridine-3′-phosphorothioate


s
phosphorothioate linkage


L10
N-(cholesterylcarboxamidocaproyl)-4-hydroxyprolinol



(Hyp-C6-Chol)


L96
N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol



(Hyp-(GalNAc-alkyl)3)


Y34
2-hydroxymethyl-tetrahydrofurane-4-methoxy-3-phosphate



(abasic 2′-OMe furanose)


Y44
inverted abasic DNA (2-hydroxymethyl-tetrahydrofurane-5-



phosphate)


(Agn)
Adenosine-glycol nucleic acid (GNA)


(Cgn)
Cytidine-glycol nucleic acid (GNA)


(Ggn)
Guanosine-glycol nucleic acid (GNA)


(Tgn)
Thymidine-glycol nucleic acid (GNA) S-Isomer


P
Phosphate


VP
Vinyl-phosphonate


dA
2′-deoxyadenosine-3′-phosphate


dAs
2′-deoxyadenosine-3′-phosphorothioate


dC
2′-deoxycytidine-3′-phosphate


dCs
2′-deoxycytidine-3′-phosphorothioate


dG
2′-deoxyguanosine-3′-phosphate


dGs
2′-deoxyguanosine-3′-phosphorothioate


dT
2′-deoxythymidine-3′-phosphate


dTs
2′-deoxythymidine-3′-phosphorothioate


dU
2′-deoxyuridine


dUs
2′-deoxyuridine-3′-phosphorothioate


(C2p)
cytidine-2′-phosphate


(G2p)
guanosine-2′-phosphate


(U2p)
uridine-2′-phosphate


(A2p)
adenosine-2′-phosphate


(Ahd)
2′-O-hexadecyl-adenosine-3′-phosphate


(Ahds)
2′-O-hexadecyl-adenosine-3′-phosphorothioate


(Chd)
2′-O-hexadecyl-cytidine-3′-phosphate


(Chds)
2′-O-hexadecyl-cytidine-3′-phosphorothioate


(Ghd)
2′-O-hexadecyl-guanosine-3′-phosphate


(Ghds)
2′-O-hexadecyl-guanosine-3′-phosphorothioate


(Uhd)
2′-O-hexadecyl-uiidinc-3′-phosphate


(Uhds)
2′-O-hexadecyl-uridine-3′-phosphorothioate
















TABLE 2







Unmodified Sense and Antisense Strand Sequences of Complement Component C3 dsRNA Agents


















SEQ





SEQ ID
Range in

ID
Range in


Duplex Name
Sense Sequence 5′ to 3′
NO:
NM_000064.3
Antisense Sequence 5′ to 3′
NO:
NM_000064.3
















AD-564727.1
CGGGUACCUCUUCAUCCAGAU
15
474-494
AUCUGGAUGAAGAGGUACCCGCU
103
472-494





AD-564730.1
GUACCUCUUCAUCCAGACAGU
16
477-497
ACUGUCTGGAUGAAGAGGUACCC
104
475-497





AD-564731.1
UACCUCUUCAUCCAGACAGAU
17
478-498
AUCUGUCUGGAUGAAGAGGUACC
105
476-498





AD-564739.1
CAUCCAGACAGACAAGACCAU
18
486-506
AUGGUCTUGUCUGUCUGGAUGAA
106
484-506





AD-564742.1
CCAGACAGACAAGACCAUCUU
19
489-509
AAGAUGGUCUUGUCUGUCUGGAU
107
487-509





AD-564744.1
AGACAGACAAGACCAUCUACU
20
491-511
AGUAGATGGUCUUGUCUGUCUGG
108
489-511





AD-564745.1
GACAGACAAGACCAUCUACAU
21
492-512
AUGUAGAUGGUCUUGUCUGUCUG
109
490-512





AD-564901.1
AUUCCGGAACUCGUCAACAUU
22
676-696
AAUGUUGACGAGUUCCGGAAUGU
110
674-696





AD-564975.1
CACUGAGUUUGAGGUGAAGGU
23
750-770
ACCUUCACCUCAAACUCAGUGGA
ill
748-770





AD-564976.1
ACUGAGUUUGAGGUGAAGGAU
24
751-771
AUCCUUCACCUCAAACUCAGUGG
112
749-771





AD-565005.1
GCCCAGUUUCGAGGUCAUAGU
25
780-800
ACUAUGACCUCGAAACUGGGCAG
113
778-800





AD-565040.1
AAUUCUACUACAUCUAUAACU
26
815-835
AGUUAUAGAUGUAGUAGAAUUUC
114
813-835





AD-565278.1
UCCCUACCAGAUCCACUUCAU
27
1146-1166
AUGAAGTGGAUCUGGUAGGGAGA
115
1144-1166





AD-565279.1
CCCUACCAGAUCCACUUCACU
28
1147-1167
AGUGAAGUGGAUCUGGUAGGGAG
116
1145-1167





AD-565281.1
CUACCAGAUCCACUUCACCAU
29
1149-1169
AUGGUGAAGUGGAUCUGGUAGGG
117
1147-1169





AD-565282.1
UACCAGAUCCACUUCACCAAU
30
1150-1170
AUUGGUGAAGUGGAUCUGGUAGG
118
1148-1170





AD-565284.1
CCAGAUCCACUUCACCAAGAU
31
1152-1172
AUCUUGGUGAAGUGGAUCUGGUA
119
1150-1172





AD-565532.1
GGGCAACUCCAACAAUUACCU
32
1440-1460
AGGUAATUGUUGGAGUUGCCCAC
120
1438-1460





AD-565534.1
GCAACUCCAACAAUUACCUGU
33
1442-1462
ACAGGUAAUUGUUGGAGUUGCCC
121
1440-1462





AD-565535.1
CAACUCCAACAAUUACCUGCU
34
1443-1463
AGCAGGTAAUUGUUGGAGUUGCC
122
1441-1463





AD-565541.1
CAACAAUUACCUGCAUCUCUU
35
1449-1469
AAGAGATGCAGGUAAUUGUUGGA
123
1447-1469





AD-565616.1
CAAGAUCCGCUACUACACCUU
36
1548-1568
AAGGUGTAGUAGCGGAUCUUGGC
124
1546-1568





AD-565904.1
CGUGCUGAAUAAGAAGAACAU
37
1902-1922
AUGUUCTUCUUAUUCAGCACGAA
125
1900-1922





AD-565905.1
GUGCUGAAUAAGAAGAACAAU
38
1903-1923
AUUGUUCUUCUUAUUCAGCACGA
126
1901-1923





AD-565925.1
ACUGACGCAGAGUAAGAUCUU
39
1923-1943
AAGAUCTUACUCUGCGUCAGUUU
127
1921-1943





AD-566234.1
UGCAGAAGAGAACAUCGUUUU
40
2361-2381
AAAACGAUGUUCUCUUCUGCAAU
128
2359-2381





AD-566383.1
CAUGUCGGACAAGAAAGGGAU
41
2517-2537
AUCCCUTUCUUGUCCGACAUGCU
129
2515-2537





AD-566384.1
AUGUCGGACAAGAAAGGGAUU
42
2518-2538
AAUCCCTUUCUUGUCCGACAUGC
130
2516-2538





AD-566386.1
GUCGGACAAGAAAGGGAUCUU
43
2520-2540
AAGAUCCCUUUCUUGUCCGACAU
131
2518-2540





AD-566388.1
CGGACAAGAAAGGGAUCUGUU
44
2522-2542
AACAGATCCCUUUCUUGUCCGAC
132
2520-2542





AD-566409.1
ACAGUAAUGCAGGACUUCUUU
45
2563-2583
AAAGAAGUCCUGCAUUACUGUGA
133
2561-2583





AD-566411.1
AGUAAUGCAGGACUUCUUCAU
46
2565-2585
AUGAAGAAGUCCUGCAUUACUGU
134
2563-2585





AD-566412.1
GUAAUGCAGGACUUCUUCAUU
47
2566-2586
AAUGAAGAAGUCCUGCAUUACUG
135
2564-2586





AD-566442.1
CUACCCUACUCUGUUGUUCGU
48
2596-2616
ACGAACAACAGAGUAGGGUAGCC
136
2594-2616





AD-566443.1
UACCCUACUCUGUUGUUCGAU
49
2597-2617
AUCGAACAACAGAGUAGGGUAGC
137
2595-2617





AD-566444.1
ACCCUACUCUGUUGUUCGAAU
50
2598-2618
AUUCGAACAACAGAGUAGGGUAG
138
2596-2618





AD-566445.1
CCCUACUCUGUUGUUCGAAAU
51
2599-2619
AUUUCGAACAACAGAGUAGGGUA
139
2597-2619





AD-566446.1
CCUACUCUGUUGUUCGAAACU
52
2600-2620
AGUUUCGAACAACAGAGUAGGGU
140
2598-2620





AD-566447.1
CUACUCUGUUGUUCGAAACGU
53
2601-2621
ACGUUUCGAACAACAGAGUAGGG
141
2599-2621





AD-566448.1
UACUCUGUUGUUCGAAACGAU
54
2602-2622
AUCGUUTCGAACAACAGAGUAGG
142
2600-2622





AD-566449.1
ACUCUGUUGUUCGAAACGAGU
55
2603-2623
ACUCGUTUCGAACAACAGAGUAG
143
2601-2623





AD-566485.1
CCGUUCUCUACAAUUACCGGU
56
2639-2659
ACCGGUAAUUGUAGAGAACGGCU
144
2637-2659





AD-566528.1
GGUGGAACUACUCCACAAUCU
57
2682-2702
AGAUUGTGGAGUAGUUCCACCCU
145
2680-2702





AD-566837.1
CCGAGUCUGAGACCAGAAUUU
58
3014-3034
AAAUUCTGGUCUCAGACUCGGUG
146
3012-3034





AD-566935.1
GUGCAUUACCUGGAUGAAACU
59
3166-3186
AGUUUCAUCCAGGUAAUGCACAG
147
3164-3186





AD-567063.1
CUACGUGGUCAAGGUCUUCUU
60
3333-3353
AAGAAGACCUUGACCACGUAGGC
148
3331-3353





AD-567066.1
CGUGGUCAAGGUCUUCUCUCU
61
3336-3356
AGAGAGAAGACCUUGACCACGUA
149
3334-3356





AD-567067.1
GUGGUCAAGGUCUUCUCUCUU
62
3337-3357
AAGAGAGAAGACCUUGACCACGU
150
3335-3357





AD-567156.1
CGUGAUACACCAAGAAAUGAU
63
3462-3482
AUCAUUTCUUGGUGUAUCACGGG
151
3460-3482





AD-567215.1
CGGCCUUUGUUCUCAUCUCGU
64
3524-3544
ACGAGATGAGAACAAAGGCCGUG
152
3522-3544





AD-567304.1
GACUUCCUUGAAGCCAACUAU
65
3613-3633
AUAGUUGGCUUCAAGGAAGUCUC
153
3611-3633





AD-567307.1
UUCCUUGAAGCCAACUACAUU
66
3616-3636
AAUGUAGUUGGCUUCAAGGAAGU
154
3614-3636





AD-567314.1
AAGCCAACUACAUGAACCUAU
67
3623-3643
AUAGGUTCAUGUAGUUGGCUUCA
155
3621-3643





AD-567315.1
AGCCAACUACAUGAACCUACU
68
3624-3644
AGUAGGTUCAUGUAGUUGGCUUC
156
3622-3644





AD-567318.1
CAACUACAUGAACCUACAGAU
69
3627-3647
AUCUGUAGGUUCAUGUAGUUGGC
157
3625-3647





AD-567395.1
UUCUGACCACAGCCAAAGAUU
70
3722-3742
AAUCUUTGGCUGUGGUCAGAAAU
158
3720-3742





AD-567487.1
UGCAGCUAAAAGACUUUGACU
71
3815-3835
AGUCAAAGUCUUUUAGCUGCAGU
159
3813-3835





AD-567521.1
CGUGCGUUGGCUCAAUGAACU
72
3849-3869
AGUUCATUGAGCCAACGCACGAC
160
3847-3869





AD-567582.1
UUCAUGGUGUUCCAAGCCUUU
73
3910-3930
AAAGGCTUGGAACACCAUGAAGG
161
3908-3930





AD-567699.1
CUGCGAUCAGAAGAGACCAAU
74
4048-4068
AUUGGUCUCUUCUGAUCGCAGGA
162
4046-4068





AD-567700.1
UGCGAUCAGAAGAGACCAAGU
75
4049-4069
ACUUGGTCUCUUCUGAUCGCAGG
163
4047-4069





AD-567713.1
ACCAAGGAAAAUGAGGGUUUU
76
4063-4083
AAAACCCUCAUUUUCCUUGGUCU
164
4061-4083





AD-567716.1
AAGGAAAAUGAGGGUUUCACU
77
4066-4086
AGUGAAACCCUCAUUUUCCUUGG
165
4064-4086





AD-567808.1
ACUCACCUGUAAUAAAUUCGU
78
4158-4178
ACGAAUTUAUUACAGGUGAGUUG
166
4156-4178





AD-567809.1
CUCACCUGUAAUAAAUUCGAU
79
4159-4179
AUCGAATUUAUUACAGGUGAGUU
167
4157-4179





AD-567812.1
ACCUGUAAUAAAUUCGACCUU
80
4162-4182
AAGGUCGAAUUUAUUACAGGUGA
168
4160-4182





AD-567813.1
CCUGUAAUAAAUUCGACCUCU
81
4163-4183
AGAGGUCGAAUUUAUUACAGGUG
169
4161-4183





AD-567814.1
CUGUAAUAAAUUCGACCUCAU
82
4164-4184
AUGAGGTCGAAUUUAUUACAGGU
170
4162-4184





AD-567828.1
ACCUCAAGGUCACCAUAAAAU
83
4178-4198
AUUUUATGGUGACCUUGAGGUCG
171
4176-4198





AD-567829.1
CCUCAAGGUCACCAUAAAACU
84
4179-4199
AGUUUUAUGGUGACCUUGAGGUC
172
4177-4199





AD-567831.1
UCAAGGUCACCAUAAAACCAU
85
4181-4201
AUGGUUTUAUGGUGACCUUGAGG
173
4179-4201





AD-568003.1
CAGAUACAUCUCCAAGUAUGU
86
4371-4391
ACAUACTUGGAGAUGUAUCUGUC
174
4369-4391





AD-568026.1
UGGACAAAGCCUUCUCCGAUU
87
4394-4414
AAUCGGAGAAGGCUUUGUCCAGC
175
4392-4414





AD-568099.1
UCUAGCUUUCAAAGUUCACCU
88
4467-4487
AGGUGAACUUUGAAAGCUAGACA
176
4465-4487





AD-568100.1
CUAGCUUUCAAAGUUCACCAU
89
4468-4488
AUGGUGAACUUUGAAAGCUAGAC
177
4466-4488





AD-568153.1
AGUCAAGGUCUACGCCUAUUU
90
4521-4541
AAAUAGGCGUAGACCUUGACUGC
178
4519-4541





AD-568156.1
CAAGGUCUACGCCUAUUACAU
91
4524-4544
AUGUAATAGGCGUAGACCUUGAC
179
4522-4544





AD-568157.1
AAGGUCUACGCCUAUUACAAU
92
4525-4545
AUUGUAAUAGGCGUAGACCUUGA
180
4523-4545





AD-568158.1
AGGUCUACGCCUAUUACAACU
93
4526-4546
AGUUGUAAUAGGCGUAGACCUUG
181
4524-4546





AD-568160.1
GUCUACGCCUAUUACAACCUU
94
4528-4548
AAGGUUGUAAUAGGCGUAGACCU
182
4526-4548





AD-568161.1
UCUACGCCUAUUACAACCUGU
95
4529-4549
ACAGGUTGUAAUAGGCGUAGACC
183
4527-4549





AD-568341.1
GGAGUGGACUAUGUGUACAAU
96
4711-4731
AUUGUACACAUAGUCCACUCCUG
184
4709-4731





AD-568343.1
AGUGGACUAUGUGUACAAGAU
97
4713-4733
AUCUUGTACACAUAGUCCACUCC
185
4711-4733





AD-568344.1
GUGGACUAUGUGUACAAGACU
98
4714-4734
AGUCUUGUACACAUAGUCCACUC
186
4712-4734





AD-568345.1
UGGACUAUGUGUACAAGACCU
99
4715-4735
AGGUCUTGUACACAUAGUCCACU
187
4713-4735





AD-568381.1
AGCUGUCCAAUGACUUUGACU
100
4751-4771
AGUCAAAGUCAUUGGACAGCUGA
188
4749-477I





AD-568382.1
GCUGUCCAAUGACUUUGACGU
101
4752-4772
ACGUCAAAGUCAUUGGACAGCUG
189
4750-4772





AD-568586.1
GAGAACCAGAAACAAUGCCAU
102
5014-5034
AUGGCATUGUUUCUGGUUCUCUU
190
5012-5034
















TABLE 3





Modified Sense and Antisense Strand Sequences of Complement Component C3 dsRNA Agents





















SEQ

SEQ


Duplex

ID

ID


Name
Sense Sequence 5′ to 3′
NO:
Antisense Sequence 5′ to 3′
NO:





AD-564727.1
csgsgguaCfcUfCfUfucauccagauL96
191
asUfscugg(Agn)ugaagaGfgUfacccgscsu
279





AD-564730.1
gsusaccuCfuUfCfAfuccagacaguL96
192
asCfsuguc(Tgn)ggaugaAfgAfgguacscsc
280





AD-564731.1
usasccucUfuCfAfUfccagacagauL96
193
asUfscugu(Cgn)uggaugAfaGfagguascsc
281





AD-564739.1
csasuccaGfaCfAfGfacaagaccauL96
194
asUfsgguc(Tgn)ugucugUfcUfggaugsasa
282





AD-564742.1
cscsagacAfgAfCfAfagaccaucuuL96
195
asAfsgaug(Ggn)ucuuguCfuGfucuggsasu
283





AD-564744.1
asgsacagAfcAfAfGfaccaucuacuL96
196
asGfsuaga(Tgn)ggucuuGfuCfugucusgsg
284





AD-564745.1
gsascagaCfaAfGfAfccaucuacauL96
197
asUfsguag(Agn)uggucuUfgUfcugucsusg
285





AD-564901.1
asusuccgGfaAfCfUfcgucaacauuL96
198
asAfsuguu(Ggn)acgaguUfcCfggaausgsu
286





AD-564975.1
csascugaGfuUfUfGfaggugaagguL96
199
asCfscuuc(Agn)ccucaaAfcUfcagugsgsa
287





AD-564976.1
ascsugagUfuUfGfAfggugaaggauL96
200
asUfsccuu(Cgn)accucaAfaCfucagusgsg
288





AD-565005.1
gscsccagUfuUfCfGfaggucauaguL96
201
asCfsuaug(Agn)ccucgaAfaCfugggcsasg
289





AD-565040.1
asasuucuAfcUfAfCfaucuauaacuL96
202
asGfsuuau(Agn)gauguaGfuAfgaauususc
290





AD-565278.1
uscsccuaCfcAfGfAfuccacuucauL96
203
asUfsgaag(Tgn)ggaucuGfgUfagggasgsa
291





AD-565279.1
cscscuacCfaGfAfUfccacuucacuL96
204
asGfsugaa(Ggn)uggaucUfgGfuagggsasg
292





AD-565281.1
csusaccaGfaUfCfCfacuucaccauL96
205
asUfsggug(Agn)aguggaUfcUfgguagsgsg
293





AD-565282.1
usasccagAfuCfCfAfcuucaccaauL96
206
asUfsuggu(Ggn)aaguggAfuCfugguasgsg
294





AD-565284.1
cscsagauCfcAfCfUfucaccaagauL96
207
asUfscuug(Ggn)ugaaguGfgAfucuggsusa
295





AD-565532.1
gsgsgcaaCfuCfCfAfacaauuaccuL96
208
asGfsguaa(Tgn)uguuggAfgUfugcccsasc
296





AD-565534.1
gscsaacuCfcAfAfCfaauuaccuguL96
209
asCfsaggu(Agn)auuguuGfgAfguugcscsc
297





AD-565535.1
csasacucCfaAfCfAfauuaccugcuL96
210
asGfscagg(Tgn)aauuguUfgGfaguugscsc
298





AD-565541.1
csasacaaUfuAfCfCfugcaucucuuL96
211
asAfsgaga(Tgn)gcagguAfaUfuguugsgsa
299





AD-565616.1
csasagauCfcGfCfUfacuacaccuuL96
212
asAfsggug(Tgn)aguagcGfgAfucuugsgsc
300





AD-565904.1
csgsugcuGfaAfUfAfagaagaacauL96
213
asUfsguuc(Tgn)ucuuauUfcAfgcacgsasa
301





AD-565905.1
gsusgcugAfaUfAfAfgaagaacaauL96
214
asUfsuguu(Cgn)uucuuaUfuCfagcacsgsa
302





AD-565925.1
ascsugacGfcAfGfAfguaagaucuuL96
215
asAfsgauc(Tgn)uacucuGfcGfucagususu
303





AD-566234.1
usgscagaAfgAfGfAfacaucguuuuL96
216
asAfsaacg(Agn)uguucuCfuUfcugcasasu
304





AD-566383.1
csasugucGfgAfCfAfagaaagggauL96
217
asUfscccu(Tgn)ucuuguCfcGfacaugscsu
305





AD-566384.1
asusgucgGfaCfAfAfgaaagggauuL96
218
asAfsuccc(Tgn)uucuugUfcCfgacausgsc
306





AD-566386.1
gsuscggaCfaAfGfAfaagggaucuuL96
219
asAfsgauc(Cgn)cuuucuUfgUfccgacsasu
307





AD-566388.1
csgsgacaAfgAfAfAfgggaucuguuL96
220
asAfscaga(Tgn)cccuuuCfuUfguccgsasc
308





AD-566409.1
ascsaguaAfuGfCfAfggacuucuuuL96
221
asAfsagaa(Ggn)uccugcAfuUfacugusgsa
309





AD-566411.1
asgsuaauGfcAfGfGfacuucuucauL96
222
asUfsgaag(Agn)aguccuGfcAfuuacusgsu
310





AD-566412.1
gsusaaugCfaGfGfAfcuucuucauuL96
223
asAfsugaa(Ggn)aaguccUfgCfauuacsusg
311





AD-566442.1
csusacccUfaCfUfCfuguuguucguL96
224
asCfsgaac(Agn)acagagUfaGfgguagscsc
312





AD-566443.1
usascccuAfcUfCfUfguuguucgauL96
225
asUfscgaa(Cgn)aacagaGfuAfggguasgsc
313





AD-566444.1
ascsccuaCfuCfUfGfuuguucgaauL96
226
asUfsucga(Agn)caacagAfgUfagggusasg
314





AD-566445.1
cscscuacUfcUfGfUfuguucgaaauL96
227
asUfsuucg(Agn)acaacaGfaGfuagggsusa
315





AD-566446.1
cscsuacuCfuGfUfUfguucgaaacuL96
228
asGfsuuuc(Ggn)aacaacAfgAfguaggsgsu
316





AD-566447.1
csusacucUfgUfUfGfuucgaaacguL96
229
asCfsguuu(Cgn)gaacaaCfaGfaguagsgsg
317





AD-566448.1
usascucuGfuUfGfUfucgaaacgauL96
230
asUfscguu(Tgn)cgaacaAfcAfgaguasgsg
318





AD-566449.1
ascsucugUfuGfUfUfcgaaacgaguL96
231
asCfsucgu(Tgn)ucgaacAfaCfagagusasg
319





AD-566485.1
cscsguucUfcUfAfCfaauuaccgguL96
232
asCfscggu(Agn)auuguaGfaGfaacggscsu
320





AD-566528.1
gsgsuggaAfcUfAfCfuccacaaucuL96
233
asGfsauug(Tgn)ggaguaGfuUfccaccscsu
321





AD-566837.1
cscsgaguCfuGfAfGfaccagaauuuL96
234
asAfsauuc(Tgn)ggucucAfgAfcucggsusg
322





AD-566935.1
gsusgcauUfaCfCfUfggaugaaacuL96
235
asGfsuuuc(Agn)uccaggUfaAfugcacsasg
323





AD-567063.1
csusacguGfgUfCfAfaggucuucuuL96
236
asAfsgaag(Agn)ccuugaCfcAfcguagsgsc
324





AD-567066.1
csgsugguCfaAfGfGfucuucucucuL96
237
asGfsagag(Agn)agaccuUfgAfccacgsusa
325





AD-567067.1
gsusggucAfaGfGfUfcuucucucuuL96
238
asAfsgaga(Ggn)aagaccUfuGfaccacsgsu
326





AD-567156.1
csgsugauAfcAfCfCfaagaaaugauL96
239
asUfscauu(Tgn)cuugguGfuAfucacgsgsg
327





AD-567215.1
csgsgccuUfuGfUfUfcucaucucguL96
240
asCfsgaga(Tgn)gagaacAfaAfggccgsusg
328





AD-567304.1
gsascuucCfuUfGfAfagccaacuauL96
241
asUfsaguu(Ggn)gcuucaAfgGfaagucsusc
329





AD-567307.1
ususccuuGfaAfGfCfcaacuacauuL96
242
asAfsugua(Ggn)uuggcuUfcAfaggaasgsu
330





AD-567314.1
asasgccaAfcUfAfCfaugaaccuauL96
243
asUfsaggu(Tgn)cauguaGfuUfggcuuscsa
331





AD-567315.1
asgsccaaCfuAfCfAfugaaccuacuL96
244
asGfsuagg(Tgn)ucauguAfgUfuggcususc
332





AD-567318.1
csasacuaCfaUfGfAfaccuacagauL96
245
asUfscugu(Agn)gguucaUfgUfaguugsgsc
333





AD-567395.1
ususcugaCfcAfCfAfgccaaagauuL96
246
asAfsucuu(Tgn)ggcuguGfgUfcagaasasu
334





AD-567487.1
usgscagcUfaAfAfAfgacuuugacuL96
247
asGfsucaa(Agn)gucuuuUfaGfcugcasgsu
335





AD-567521.1
csgsugcgUfuGfGfCfucaaugaacuL96
248
asGfsuuca(Tgn)ugagccAfaCfgcacgsasc
336





AD-567582.1
ususcaugGfuGfUfUfccaagccuuuL96
249
asAfsaggc(Tgn)uggaacAfcCfaugaasgsg
337





AD-567699.1
csusgcgaUfcAfGfAfagagaccaauL96
250
asUfsuggu(Cgn)ucuucuGfaUfcgcagsgsa
338





AD-567700.1
usgscgauCfaGfAfAfgagaccaaguL96
251
asCfsuugg(Tgn)cucuucUfgAfucgcasgsg
339





AD-567713.1
ascscaagGfaAfAfAfugaggguuuuL96
252
asAfsaacc(Cgn)ucauuuUfcCfuugguscsu
340





AD-567716.1
asasggaaAfaUfGfAfggguuucacuL96
253
asGfsugaa(Agn)cccucaUfuUfuccuusgsg
341





AD-567808.1
ascsucacCfuGfUfAfauaaauucguL96
254
asCfsgaau(Tgn)uauuacAfgGfugagususg
342





AD-567809.1
csuscaccUfgUfAfAfuaaauucgauL96
255
asUfscgaa(Tgn)uuauuaCfaGfgugagsusu
343





AD-567812.1
ascscuguAfaUfAfAfauucgaccuuL96
256
asAfsgguc(Ggn)aauuuaUfuAfcaggusgsa
344





AD-567813.1
cscsuguaAfuAfAfAfuucgaccucuL96
257
asGfsaggu(Cgn)gaauuuAfuUfacaggsusg
345





AD-567814.1
csusguaaUfaAfAfUfucgaccucauL96
258
asUfsgagg(Tgn)cgaauuUfaUfuacagsgsu
346





AD-567828.1
ascscucaAfgGfUfCfaccauaaaauL96
259
asUfsuuua(Tgn)ggugacCfuUfgagguscsg
347





AD-567829.1
cscsucaaGfgUfCfAfccauaaaacuL96
260
asGfsuuuu(Agn)uggugaCfcUfugaggsusc
348





AD-567831.1
uscsaaggUfcAfCfCfauaaaaccauL96
261
asUfsgguu(Tgn)uaugguGfaCfcuugasgsg
349





AD-568003.1
csasgauaCfaUfCfUfccaaguauguL96
262
asCfsauac(Tgn)uggagaUfgUfaucugsusc
350





AD-568026.1
usgsgacaAfaGfCfCfuucuccgauuL96
263
asAfsucgg(Agn)gaaggcUfuUfguccasgsc
351





AD-568099.1
uscsuagcUfuUfCfAfaaguucaccuL96
264
asGfsguga(Agn)cuuugaAfaGfcuagascsa
352





AD-568100.1
csusagcuUfuCfAfAfaguucaccauL96
265
asUfsggug(Agn)acuuugAfaAfgcuagsasc
353





AD-568153.1
asgsucaaGfgUfCfUfacgccuauuuL96
266
asAfsauag(Ggn)cguagaCfcUfugacusgsc
354





AD-568156.1
csasagguCfuAfCfGfccuauuacauL96
267
asUfsguaa(Tgn)aggcguAfgAfccuugsasc
355





AD-568157.1
asasggucUfaCfGfCfcuauuacaauL96
268
asUfsugua(Agn)uaggcgUfaGfaccuusgsa
356





AD-568158.1
asgsgucuAfcGfCfCfuauuacaacuL96
269
asGfsuugu(Agn)auaggcGfuAfgaccususg
357





AD-568160.1
gsuscuacGfcCfUfAfuuacaaccuuL96
270
asAfsgguu(Ggn)uaauagGfcGfuagacscsu
358





AD-568161.1
uscsuacgCfcUfAfUfuacaaccuguL96
271
asCfsaggu(Tgn)guaauaGfgCfguagascsc
359





AD-568341.1
gsgsagugGfaCfUfAfuguguacaauL96
272
asUfsugua(Cgn)acauagUfcCfacuccsusg
360





AD-568343.1
asgsuggaCfuAfUfGfuguacaagauL96
273
asUfscuug(Tgn)acacauAfgUfccacuscsc
361





AD-568344.1
gsusggacUfaUfGfUfguacaagacuL96
274
asGfsucuu(Ggn)uacacaUfaGfuccacsusc
362





AD-568345.1
usgsgacuAfuGfUfGfuacaagaccuL96
275
asGfsgucu(Tgn)guacacAfuAfguccascsu
363





AD-568381.1
asgscuguCfcAfAfUfgacuuugacuL96
276
asGfsucaa(Agn)gucauuGfgAfcagcusgsa
364





AD-568382.1
gscsugucCfaAfUfGfacuuugacguL96
277
asCfsguca(Agn)agucauUfgGfacagcsusg
365





AD-568586.1
gsasgaacCfaGfAfAfacaaugccauL96
278
asUfsggca(Tgn)uguuucUfgGfuucucsusu
366









SEQ




Duplex

ID




Name
mRNA target sequence
NO:







AD-564727.1
AGCGGGUACCUCUUCAUCCAGAC
367







AD-564730.1
GGGUACCUCUUCAUCCAGACAGA
368







AD-564731.1
GGUACCUCUUCAUCCAGACAGAC
369







AD-564739.1
UUCAUCCAGACAGACAAGACCAU
370







AD-564742.1
AUCCAGACAGACAAGACCAUCUA
371







AD-564744.1
CCAGACAGACAAGACCAUCUACA
372







AD-564745.1
CAGACAGACAAGACCAUCUACAC
373







AD-564901.1
ACAUUCCGGAACUCGUCAACAUG
374







AD-564975.1
UCCACUGAGUUUGAGGUGAAGGA
375







AD-564976.1
CCACUGAGUUUGAGGUGAAGGAG
376







AD-565005.1
CUGCCCAGUUUCGAGGUCAUAGU
377







AD-565040.1
GAAAUUCUACUACAUCUAUAACG
378







AD-565278.1
UCUCCCUACCAGAUCCACUUCAC
379







AD-565279.1
CUCCCUACCAGAUCCACUUCACC
380







AD-565281.1
CCCUACCAGAUCCACUUCACCAA
381







AD-565282.1
CCUACCAGAUCCACUUCACCAAG
382







AD-565284.1
UACCAGAUCCACUUCACCAAGAC
383







AD-565532.1
GUGGGCAACUCCAACAAUUACCU
384







AD-565534.1
GGGCAACUCCAACAAUUACCUGC
385







AD-565535.1
GGCAACUCCAACAAUUACCUGCA
386







AD-565541.1
UCCAACAAUUACCUGCAUCUCUC
387







AD-565616.1
GCCAAGAUCCGCUACUACACCUA
388







AD-565904.1
UUCGUGCUGAAUAAGAAGAACAA
389







AD-565905.1
UCGUGCUGAAUAAGAAGAACAAA
390







AD-565925.1
AAACUGACGCAGAGUAAGAUCUG
391







AD-566234.1
AUUGCAGAAGAGAACAUCGUUUC
392







AD-566383.1
AGCAUGUCGGACAAGAAAGGGAU
393







AD-566384.1
GCAUGUCGGACAAGAAAGGGAUC
394







AD-566386.1
AUGUCGGACAAGAAAGGGAUCUG
395







AD-566388.1
GUCGGACAAGAAAGGGAUCUGUG
396







AD-566409.1
UCACAGUAAUGCAGGACUUCUUC
397







AD-566411.1
ACAGUAAUGCAGGACUUCUUCAU
398







AD-566412.1
CAGUAAUGCAGGACUUCUUCAUC
399







AD-566442.1
GGCUACCCUACUCUGUUGUUCGA
400







AD-566443.1
GCUACCCUACUCUGUUGUUCGAA
401







AD-566444.1
CUACCCUACUCUGUUGUUCGAAA
402







AD-566445.1
UACCCUACUCUGUUGUUCGAAAC
403







AD-566446.1
ACCCUACUCUGUUGUUCGAAACG
404







AD-566447.1
CCCUACUCUGUUGUUCGAAACGA
405







AD-566448.1
CCUACUCUGUUGUUCGAAACGAG
406







AD-566449.1
CUACUCUGUUGUUCGAAACGAGC
407







AD-566485.1
AGCCGUUCUCUACAAUUACCGGC
408







AD-566528.1
AGGGUGGAACUACUCCACAAUCC
409







AD-566837.1
CACCGAGUCUGAGACCAGAAUUC
410







AD-566935.1
CUGUGCAUUACCUGGAUGAAACG
411







AD-567063.1
GCCUACGUGGUCAAGGUCUUCUC
412







AD-567066.1
UACGUGGUCAAGGUCUUCUCUCU
413







AD-567067.1
ACGUGGUCAAGGUCUUCUCUCUG
414







AD-567156.1
CCCGUGAUACACCAAGAAAUGAU
415







AD-567215.1
CACGGCCUUUGUUCUCAUCUCGC
416







AD-567304.1
GAGACUUCCUUGAAGCCAACUAC
417







AD-567307.1
ACUUCCUUGAAGCCAACUACAUG
418







AD-567314.1
UGAAGCCAACUACAUGAACCUAC
419







AD-567315.1
GAAGCCAACUACAUGAACCUACA
420







AD-567318.1
GCCAACUACAUGAACCUACAGAG
421







AD-567395.1
AUUUCUGACCACAGCCAAAGAUA
422







AD-567487.1
ACUGCAGCUAAAAGACUUUGACU
423







AD-567521.1
GUCGUGCGUUGGCUCAAUGAACA
424







AD-567582.1
CCUUCAUGGUGUUCCAAGCCUUG
425







AD-567699.1
UCCUGCGAUCAGAAGAGACCAAG
426







AD-567700.1
CCUGCGAUCAGAAGAGACCAAGG
427







AD-567713.1
AGACCAAGGAAAAUGAGGGUUUC
428







AD-567716.1
CCAAGGAAAAUGAGGGUUUCACA
429







AD-567808.1
CAACUCACCUGUAAUAAAUUCGA
430







AD-567809.1
AACUCACCUGUAAUAAAUUCGAC
431







AD-567812.1
UCACCUGUAAUAAAUUCGACCUC
432







AD-567813.1
CACCUGUAAUAAAUUCGACCUCA
433







AD-567814.1
ACCUGUAAUAAAUUCGACCUCAA
434







AD-567828.1
CGACCUCAAGGUCACCAUAAAAC
435







AD-567829.1
GACCUCAAGGUCACCAUAAAACC
436







AD-567831.1
CCUCAAGGUCACCAUAAAACCAG
437







AD-568003.1
GACAGAUACAUCUCCAAGUAUGA
438







AD-568026.1
GCUGGACAAAGCCUUCUCCGAUA
439







AD-568099.1
UGUCUAGCUUUCAAAGUUCACCA
440







AD-568100.1
GUCUAGCUUUCAAAGUUCACCAA
441







AD-568153.1
GCAGUCAAGGUCUACGCCUAUUA
442







AD-568156.1
GUCAAGGUCUACGCCUAUUACAA
443







AD-568157.1
UCAAGGUCUACGCCUAUUACAAC
444







AD-568158.1
CAAGGUCUACGCCUAUUACAACC
445







AD-568160.1
AGGUCUACGCCUAUUACAACCUG
446







AD-568161.1
GGUCUACGCCUAUUACAACCUGG
447







AD-568341.1
CAGGAGUGGACUAUGUGUACAAG
448







AD-568343.1
GGAGUGGACUAUGUGUACAAGAC
449







AD-568344.1
GAGUGGACUAUGUGUACAAGACC
450







AD-568345.1
AGUGGACUAUGUGUACAAGACCC
451







AD-568381.1
UCAGCUGUCCAAUGACUUUGACG
452







AD-568382.1
CAGCUGUCCAAUGACUUUGACGA
453







AD-568586.1
AAGAGAACCAGAAACAAUGCCAG
454
















TABLE 4







Unmodified Sense and Antisense Strand Sequences of Complement Component C3 dsRNA Agents















SEQ


SEQ





ID
Range in

ID
Range in


Duplex Name
Sense Sequence 5′ to 3′
NO:
NM_000064.3
Antisense Sequence 5′ to 3′
NO:
NM_000064.3





AD-569034.1
ACGGUCAUGGUCAACAUUGAU
455
577-597
AUCAAUGUUGACCAUGACCGUCC
489
575-597





AD-569164.1
AGAUCCGAGCCUACUAUGAAU
456
707-727
AUUCAUAGUAGGCUCGGAUCUUC
490
705-727





AD-569165.1
GAUCCGAGCCUACUAUGAAAU
457
708-728
AUUUCAUAGUAGGCUCGGAUCUU
491
706-728





AD-569272.1
AAUUCUACUACAUCUAUAACU
458
815-835
AGUUAUAGAUGUAGUAGAAUUUC
492
813-835





AD-569763.1
UGGGCAACUCCAACAAUUACU
459
1439-1459
AGUAAUUGUUGGAGUUGCCCACG
493
1437-1459





AD-569765.1
GGCAACUCCAACAAUUACCUU
460
1441-1461
AAGGUAAUUGUUGGAGUUGCCCA
494
1439-1461





AD-570130.1
CGUGUUCGUGCUGAAUAAGAU
461
1896-1916
AUCUUAUUCAGCACGAACACGCC
495
1894-1916





AD-570132.1
UGUUCGUGCUGAAUAAGAAGU
462
1898-1918
ACUUCUUAUUCAGCACGAACACG
496
1896-1918





AD-570133.1
GUUCGUGCUGAAUAAGAAGAU
463
1899-1919
AUCUUCUUAUUCAGCACGAACAC
497
1897-1919





AD-570134.1
UUCGUGCUGAAUAAGAAGAAU
464
1900-1920
AUUCUUCUUAUUCAGCACGAACA
498
1898-1920





AD-570157.1
ACUGACGCAGAGUAAGAUCUU
465
1923-1943
AAGAUCUUACUCUGCGUCAGUUU
499
1921-1943





AD-570711.1
UCCGAGCCGUUCUCUACAAUU
466
2633-2653
AAUUGUAGAGAACGGCUCGGAUU
500
2631-2653





AD-570712.1
CCGAGCCGUUCUCUACAAUUU
467
2634-2654
AAAUUGUAGAGAACGGCUCGGAU
501
2632-2654





AD-570713.1
CGAGCCGUUCUCUACAAUUAU
468
2635-2655
AUAAUUGUAGAGAACGGCUCGGA
502
2633-2655





AD-570714.1
GAGCCGUUCUCUACAAUUACU
469
2636-2656
AGUAAUUGUAGAGAACGGCUCGG
503
2634-2656





AD-571539.1
UUCCUUGAAGCCAACUACAUU
470
3616-3636
AAUGUAGUUGGCUUCAAGGAAGU
504
3614-3636





AD-571610.1
GCCUCUUCUUAACAAAUUUCU
471
3705-3725
AGAAAUUUGUUAAGAAGAGGCCC
505
3703-3725





AD-571633.1
CCACAGCCAAAGAUAAGAACU
472
3728-3748
AGUUCUUAUCUUUGGCUGUGGUC
506
3726-3748





AD-571715.1
CUACUGCAGCUAAAAGACUUU
473
3811-3831
AAAGUCUUUUAGCUGCAGUAGGG
507
3809-3831





AD-571752.1
UCGUGCGUUGGCUCAAUGAAU
474
3848-3868
AUUCAUUGAGCCAACGCACGACG
508
3846-3868





AD-571754.1
GUGCGUUGGCUCAAUGAACAU
475
3850-3870
AUGUUCAUUGAGCCAACGCACGA
509
3848-3870





AD-571828.1
AGCCUUGGCUCAAUACCAAAU
476
3924-3944
AUUUGGUAUUGAGCCAAGGCUUG
510
3922-3944





AD-572039.1
AACUCACCUGUAAUAAAUUCU
477
4157-4177
AGAAUUUAUUACAGGUGAGUUGA
511
4155-4177





AD-572040.1
ACUCACCUGUAAUAAAUUCGU
478
4158-4178
ACGAAUUUAUUACAGGUGAGUUG
512
4156-4178





AD-572041.1
CUCACCUGUAAUAAAUUCGAU
479
4159-4179
AUCGAAUUUAUUACAGGUGAGUU
513
4157-4179





AD-572059.1
GACCUCAAGGUCACCAUAAAU
480
4177-4197
AUUUAUGGUGACCUUGAGGUCGA
514
4175-4197





AD-572061.1
CCUCAAGGUCACCAUAAAACU
481
4179-4199
AGUUUUAUGGUGACCUUGAGGUC
515
4177-4199





AD-572062.1
CUCAAGGUCACCAUAAAACCU
482
4180-4200
AGGUUUUAUGGUGACCUUGAGGU
516
4178-4200





AD-572063.1
UCAAGGUCACCAUAAAACCAU
483
4181-4201
AUGGUUUUAUGGUGACCUUGAGG
517
4179-4201





AD-572110.1
GAUGCCAAGAACACUAUGAUU
484
4228-4248
AAUCAUAGUGUUCUUGGCAUCCU
518
4226-4248





AD-572144.1
AGGAUGCCACUAUGUCUAUAU
485
4280-4300
AUAUAGACAUAGUGGCAUCCUGG
519
4278-4300





AD-572388.1
CAAGGUCUACGCCUAUUACAU
486
4524-4544
AUGUAAUAGGCGUAGACCUUGAC
520
4522-4544





AD-572389.1
AAGGUCUACGCCUAUUACAAU
487
4525-4545
AUUGUAAUAGGCGUAGACCUUGA
521
4523-4545





AD-572390.1
AGGUCUACGCCUAUUACAACU
488
4526-4546
AGUUGUAAUAGGCGUAGACCUUG
522
4524-4546
















TABLE 5







Modified Sense and Antisense Strand Sequences


of Complement Component C3 dsRNA Agents














Sense
SEQ
Antisense
SEQ
mRNA
SEQ


Duplex
Sequence
ID
Sequence
ID
target
ID


Name
5′ to 3′
NO:
5′ to 3′
NO:
sequence
NO:





AD-
ascsgg
523
asUfsc
557
GGACGG
591


569034.1
ucAfuG

aaUfgU

UCAUGG




fGfUfc

fUfgac

UCAACA




aacauu

cAfuGf

UUGAG




gauL96

accgus








csc








AD-
asgsau
524
asUfsu
558
GAAGAU
592


569164.1
ccGfaG

caUfaG

CCGAGC




fCfCfu

fUfagg

CUACUA




acuaug

cUfcGf

UGAAA




aauL96

gaucus








usc








AD-
gsasuc
525
asUfsu
559
AAGAUC
593


569165.1
cgAfgC

ucAfuA

CGAGCC




fCfUfa

fGfuag

UACUAU




cuauga

gCfuCf

GAAAA




aauL96

ggaucs








usu








AD-
asasuu
526
asGfsu
560
GAAAUU
594


569272.1
cuAfcU

uaUfaG

CUACUA




fAfCfa

fAfugu

CAUCUA




ucuaua

aGfuAf

UAACG




acuL96

gaauus








usc








AD-
usgsgg
527
asGfsu
561
CGUGGG
595


569763.1
caAfcU

aaUfuG

CAACUC




fCfCfa

fUfugg

CAACAA




acaauu

aGfuUf

UUACC




acuL96

gcccas








csg








AD-
gsgsca
528
asAfsg
562
UGGGCA
596


569765.1
acUfcC

guAfaU

ACUCCA




fAfAfc

fUfguu

ACAAUU




aauuac

gGfaGf

ACCUG




cuuL96

uugccs








csa








AD-
csgsug
529
asUfsc
563
GGCGUG
597


570130.1
uuCfgU

uuAfuU

UUCGUG




fGfCfu

fCfagc

CUGAAU




gaauaa

aCfgAf

AAGAA




gauL96

acacgs








csc








AD-
usgsuu
530
asCfsu
564
CGUGUU
598


570132.1
cgUfgC

ucUfuA

CGUGCU




fUfGfa

fUfuca

GAAUAA




auaaga

gCfaCf

GAAGA




aguL96

gaacas








csg








AD-
gsusuc
531
asUfsc
565
GUGUUC
599


570133.1
guGfcU

uuCfuU

GUGCUG




fGfAfa

fAfuuc

AAUAAG




uaagaa

aGfcAf

AAGAA




gauL96

cgaacs








asc








AD-
ususcg
532
asUfsu
566
UGUUCG
600


570134.1
ugCfuG

cuUfcU

UGCUGA




fAfAfu

fUfauu

AUAAGA




aagaag

cAfgCf

AGAAC




aauL96

acgaas








csa








AD-
ascsug
533
asAfsg
567
AAACUG
601


570157.1
acGfcA

auCfuU

ACGCAG




fGfAfg

fAfcuc

AGUAAG




uaagau

uGfcGf

AUCUG




cuuL96

ucagus








usu








AD-
uscscg
534
asAfsu
568
AAUCCG
602


570711.1
agCfcG

ugUfaG

AGCCGU




fUfUfc

fAfgaa

UCUCUA




ucuaca

cGfgCf

CAAUU




auuL96

ucggas








usu








AD-
cscsga
535
asAfsa
569
AUCCGA
603


570712.1
gcCfgU

uuGfuA

GCCGUU




fUfCfu

fGfaga

CUCUAC




cuacaa

aCfgGf

AAUUA




uuuL96

cucggs








asu








AD-
csgsag
536
asUfsa
570
UCCGAG
604


570713.1
ccGfuU

auUfgU

CCGUUC




fCfUfc

fAfgag

UCUACA




uacaau

aAfcGf

AUUAC




uauL96

gcucgs








gsa





Ftablel.l
gsasgc
537
asGfsu
571
CCGAGC
605



cgUfuC

aaUfuG

CGUUCU




fUfCfu

fUfaga

CUACAA




acaauu

gAfaCf

UUACC




acuL96

ggcucs








gsg








AD-
ususcc
538
asAfsu
572
ACUUCC
606


571539.1
uuGfaA

guAfgU

UUGAAG




fGfCfc

fUfggc

CCAACU




aacuac

uUfcAf

ACAUG




auuL96

aggaas








gsu








AD-
gscscu
539
asGfsa
573
GGGCCU
607


571610.1
cuUfcU

aaUfuU

CUUCUU




fUfAfa

fGfuua

AACAAA




caaauu

aGfaAf

UUUCU




ucuL96

gaggcs








csc








AD-
cscsac
540
asGfsu
574
GACCAC
608


571633.1
agCfcA

ucUfuA

AGCCAA




fAfAfg

fUfcuu

AGAUAA




auaaga

uGfgCf

GAACC




acuL96

uguggs








usc








AD-
csusac
541
asAfsa
575
CCCUAC
609


571715.1
ugCfaG

guCfuU

UGCAGC




fCfUfa

fUfuag

UAAAAG




aaagac

cUfgCf

ACUUU




uuuL96

aguags








gsg








AD-
uscsgu
542
asUfsu
576
CGUCGU
610


571752.1
gcGfuU

caUfuG

GCGUUG




fGfGfc

fAfgcc

GCUCAA




ucaaug

aAfcGf

UGAAC




aauL96

cacgas








csg








AD-
gsusgc
543
asUfsg
577
UCGUGC
611


571754.1
guUfgG

uuCfaU

GUUGGC




fCfUfc

fUfgag

UCAAUG




aaugaa

cCfaAf

AACAG




cauL96

cgcacs








gsa








AD-
asgscc
544
asUfsu
578
CAAGCC
612


571828.1
uuGfgC

ugGfuA

UUGGCU




fUfCfa

fUfuga

CAAUAC




auacca

gCfcAf

CAAAA




aauL96

aggcus








usg








AD-
asascu
545
asGfsa
579
UCAACU
613


572039.1
caCfcU

auUfuA

CACCUG




fGfUfa

fUfuac

UAAUAA




auaaau

aGfgUf

AUUCG




ucuL96

gaguus








gsa








AD-
ascsuc
546
asCfsg
580
CAACUC
614


572040.1
acCfuG

aaUfuU

ACCUGU




fUfAfa

fAfuua

AAUAAA




uaaauu

cAfgGf

UUCGA




cguL96

ugagus








usg








AD-
csusca
547
asUfsc
581
AACUCA
615


572041.1
ccUfgU

gaAfuU

CCUGUA




fAfAfu

fUfauu

AUAAAU




aaauuc

aCfaGf

UCGAC




gauL96

gugags








usu








AD-
gsascc
548
asUfsu
582
UCGACC
616


572059.1
ucAfaG

uaUfgG

UCAAGG




fGfUfc

fUfgac

UCACCA




accaua

cUfuGf

UAAAA




aauL96

aggucs








gsa








AD-
cscsuc
549
asGfsu
583
GACCUC
617


572061.1
aaGfgU

uuUfaU

AAGGUC




fCfAfc

fGfgug

ACCAUA




cauaaa

aCfcUf

AAACC




acuL96

ugaggs








usc








AD-
csusca
550
asGfsg
584
ACCUCA
618


572062.1
agGfuC

uuUfuA

AGGUCA




fAfCfc

fUfggu

CCAUAA




auaaaa

gAfcCf

AACCA




ccuL96

uugags








gsu








AD-
uscsaa
551
asUfsg
585
CCUCAA
619


572063.1
ggUfcA

guUfuU

GGUCAC




fCfCfa

fAfugg

CAUAAA




uaaaac

uGfaCf

ACCAG




cauL96

cuugas








gsg








AD-
gsasug
552
asAfsu
586
AGGAUG
620


572110.1
ccAfaG

caUfaG

CCAAGA




fAfAfc

fUfguu

ACACUA




acuaug

cUfuGf

UGAUC




auuL96

gcaucs








csu








AD-
asgsga
553
asUfsa
587
CCAGGA
621


572144.1
ugCfcA

uaGfaC

UGCCAC




fCfUfa

fAfuag

UAUGUC




ugucua

uGfgCf

UAUAU




uauL96

auccus








gsg








AD-
csasag
554
asUfsg
588
GUCAAG
622


572388.1
guCfuA

uaAfuA

GUCUAC




fCfGfc

fGfgcg

GCCUAU




cuauua

uAfgAf

UACAA




cauL96

ccuugs








asc








AD-
asasgg
555
asUfsu
589
UCAAGG
623


572389.1
ucUfaC

guAfaU

UCUACG




fGfCfc

fAfggc

CCUAUU




uauuac

gUfaGf

ACAAC




aauL96

accuus








gsa








AD-
asgsgu
556
asGfsu
590
CAAGGU
624


572390.1
cuAfcG

ugUfaA

CUACGC




fCfCfu

fUfagg

CUAUUA




auuaca

cGfuAf

CAACC




acuL96

gaccus








usg
















TABLE 6







Unmodified Sense and Antisense Strand


Sequences of Complement Component C3


dsRNA Agents

















Anti-





Sense


sense





Se-

Range
Se-

Range



quence
SEQ
in
quence
SEQ
in


Duplex
5′ to
ID
NM_00
5′ to
ID
NM_00


Name
3′
NO:
0064.3
3′
NO:
0064.3
















AD-
AGACAG
625
491-511
AGUAGA
714
489-511


568976.1
ACAAGA


UGGUCU





CCAUCU


UGUCUG





ACU


UCUGG







AD-
ACAGAC
626
493-513
AGUGUA
715
491-513


568978.1
AAGACC


GAUGGU





AUCUAC


CUUGUC





ACU


UGUCU







AD-
UGGGAC
627
670-690
AACGAG
716
668-690


569127.1
AUUCCG


UUCCGG





GAACUC


AAUGUC





GUU


CCAAG







AD-
AUUCCG
628
676-696
AAUGUU
717
674-696


569133.1
GAACUC


GACGAG





GUCAAC


UUCCGG





AUU


AAUGU







AD-
AGAUCC
629
707-727
AUUCAU
718
705-727


569164.1
GAGCCU


AGUAGG





ACUAUG


CUCGGA





AAU


UCUUC







AD-
GCAGGU
630
738-758
AACUCA
719
736-758


569195.1
CUUCUC


GUGGAG





CACUGA


AAGACC





GUU


UGCUG







AD-
GCCCAG
631
780-800
ACUAUG
720
778-800


569237.1
UUUCGA


ACCUCG





GGUCAU


AAACUG





AGU


GGCAG







AD-
CCAGUU
632
782-802
ACACUA
721
780-802


569239.1
UCGAGG


UGACCU





UCAUAG


CGAAAC





UGU


UGGGC







AD-
AAUUCU
633
815-835
AGUUAU
722
813-835


569272.1
ACUACA


AGAUGU





UCUAUA


AGUAGA





ACU


AUUUC







AD-
ACUGCC
634
895-915
ACCGAA
723
893-915


569350.1
UUUGUC


GAUGAC





AUCUUC


AAAGGC





GGU


AGUUC







AD-
CUCAUG
635
1207-1227
AUUCGU
724
1205-1227


569571.1
GUGUUC


CACGAA





GUGACG


CACCAU





AAU


GAGGU







AD-
UGGGCA
636
1439-1459
AGUAAU
725
1437-1459


569763.1
ACUCCA


UGUUGG





ACAAUU


AGUUGC





ACU


CCACG







AD-
GGGCAA
637
1440-1460
AGGUAA
726
1438-1460


569764.1
CUCCAA


UUGUUG





CAAUUA


GAGUUG





CCU


CCCAC







AD-
GCAACU
638
1442-1462
ACAGGU
727
1440-1462


569766.1
CCAACA


AAUUGU





AUUACC


UGGAGU





UGU


UGCCC







AD-
GUCAAC
639
1510-1530
AAUUCG
728
1508-1530


569816.1
UUCCUC


CAGGAG





CUGCGA


GAAGUU





AUU


GACGU







AD-
AACUGA
640
1922-1942
AGAUCU
729
1920-1942


570156.1
CGCAGA


UACUCU





GUAAGA


GCGUCA





UCU


GUUUG







AD-
UGCAGA
641
2361-2381
AAAACG
730
2359-2381


570466.1
AGAGAA


AUGUUC





CAUCGU


UCUUCU





UUU


GCAAU







AD-
GAAGAG
642
2365-2385
ACGGGA
731
2363-2385


570470.1
AACAUC


AACGAU





GUUUCC


GUUCUC





CGU


UUCUG







AD-
AAGAGA
643
2366-2386
AUCGGG
732
2364-2386


570471.1
ACAUCG


AAACGA





UUUCCC


UGUUCU





GAU


CUUCU







AD-
AGAACA
644
2369-2389
AACUUC
733
2367-2389


570474.1
UCGUUU


GGGAAA





CCCGAA


CGAUGU





GUU


UCUCU







AD-
GAACAU
645
2370-2390
ACACUU
734
2368-2390


570475.1
CGUUUC


CGGGAA





CCGAAG


ACGAUG





UGU


UUCUC







AD-
AACAUC
646
2371-2391
AUCACU
735
2369-2391


570476.1
GUUUCC


UCGGGA





CGAAGU


AACGAU





GAU


GUUCU







AD-
CGGACA
647
2522-2542
AACAGA
736
2520-2542


570620.1
AGAAAG


UCCCUU





GGAUCU


UCUUGU





GUU


CCGAC







AD-
GGACAA
648
2523-2543
ACACAG
737
2521-2543


570621.1
GAAAGG


AUCCCU





GAUCUG


UUCUUG





UGU


UCCGA







AD-
GACAAG
649
2524-2544
AACACA
738
2522-2544


570622.1
AAAGGG


GAUCCC





AUCUGU


UUUCUU





GUU


GUCCG







AD-
ACAAGA
650
2525-2545
ACACAC
739
2523-2545


570623.1
AAGGGA


AGAUCC





UCUGUG


CUUUCU





UGU


UGUCC







AD-
CAAGAA
651
2526-2546
ACCACA
740
2524-2546


570624.1
AGGGAU


CAGAUC





CUGUGU


CCUUUC





GGU


UUGUC







AD-
AAGAAA
652
2527-2547
AGCCAC
741
2525-2547


570625.1
GGGAUC


ACAGAU





UGUGUG


CCCUUU





GCU


CUUGU







AD-
GAAAGG
653
2529-2549
ACUGCC
742
2527-2549


570627.1
GAUCUG


ACACAG





UGUGGC


AUCCCU





AGU


UUCUU







AD-
CUUCGA
654
2553-2573
AGCAUU
743
2551-2573


570631.1
GGUCAC


ACUGUG





AGUAAU


ACCUCG





GCU


AAGGG







AD-
UUCGAG
655
2554-2574
AUGCAU
744
2552-2574


570632.1
GUCACA


UACUGU





GUAAUG


GACCUC





CAU


GAAGG







AD-
GGCUAC
656
2594-2614
AAACAA
745
2592-2614


570672.1
CCUACU


CAGAGU





CUGUUG


AGGGUA





UUU


GCCGC







AD-
CUACCC
657
2596-2616
ACGAAC
746
2594-2616


570674.1
UACUCU


AACAGA





GUUGUU


GUAGGG





CGU


UAGCC







AD-
UACCCU
658
2597-2617
AUCGAA
747
2595-2617


570675.1
ACUCUG


CAACAG





UUGUUC


AGUAGG





GAU


GUAGC







AD-
ACCCUA
659
2598-2618
AUUCGA
748
2596-2618


570676.1
CUCUGU


ACAACA





UGUUCG


GAGUAG





AAU


GGUAG







AD-
CCCUAC
660
2599-2619
AUUUCG
749
2597-2619


570677.1
UCUGUU


AACAAC





GUUCGA


AGAGUA





AAU


GGGUA







AD-
CCUACU
661
2600-2620
AGUUUC
750
2598-2620


570678.1
CUGUUG


GAACAA





UUCGAA


CAGAGU





ACU


AGGGU







AD-
CUACUC
662
2601-2621
ACGUUU
751
2599-2621


570679.1
UGUUGU


CGAACA





UCGAAA


ACAGAG





CGU


UAGGG







AD-
UACUCU
663
2602-2622
AUCGUU
752
2600-2622


570680.1
GUUGUU


UCGAAC





CGAAAC


AACAGA





GAU


GUAGG







AD-
ACUCUG
664
2603-2623
ACUCGU
753
2601-2623


570681.1
UUGUUC


UUCGAA





GAAACG


CAACAG





AGU


AGUAG







AD-
CUCUGU
665
2604-2624
AGCUCG
754
2602-2624


570682.1
UGUUCG


UUUCGA





AAACGA


ACAACA





GCU


GAGUA







AD-
CCGUUC
666
2639-2659
ACCGGU
755
2637-2659


570717.1
UCUACA


AAUUGU





AUUACC


AGAGAA





GGU


CGGCU







AD-
AACAAA
667
2908-2928
ACGAAC
756
2906-2928


570963.1
ACUGUG


AGCCAC





GCUGUU


AGUUUU





CGU


GUUCA







AD-
GGUCAU
668
3156-3176
AGGUAA
757
3154-3176


571157.1
CGCUGU


UGCACA





GCAUUA


GCGAUG





CCU


ACCGU







AD-
GUCAUC
669
3157-3177
AAGGUA
758
3155-3177


571158.1
GCUGUG


AUGCAC





CAUUAC


AGCGAU





CUU


GACCG







AD-
UGCAUU
670
3167-3187
ACGUUU
759
3165-3187


571168.1
ACCUGG


CAUCCA





AUGAAA


GGUAAU





CGU


GCACA







AD-
CGUGGU
671
3336-3356
AGAGAG
760
3334-3356


571298.1
CAAGGU


AAGACC





CUUCUC


UUGACC





UCU


ACGUA







AD-
CGGCCU
672
3524-3544
ACGAGA
761
3522-3544


571447.1
UUGUUC


UGAGAA





UCAUCU


CAAAGG





CGU


CCGUG







AD-
GGCCUU
673
3525-3545
AGCGAG
762
3523-3545


571448.1
UGUUCU


AUGAGA





CAUCUC


ACAAAG





GCU


GCCGU







AD-
GCCUUU
674
3526-3546
AAGCGA
763
3524-3546


571449.1
GUUCUC


GAUGAG





AUCUCG


AACAAA





CUU


GGCCG







AD-
UUCCUU
675
3616-3636
AAUGUA
764
3614-3636


571539.1
GAAGCC


GUUGGC





AACUAC


UUCAAG





AUU


GAAGU







AD-
UGCAGC
676
3815-3835
AGUCAA
765
3813-3835


571719.1
UAAAAG


AGUCUU





ACUUUG


UUAGCU





ACU


GCAGU







AD-
UCGUGC
677
3848-3868
AUUCAU
766
3846-3868


571752.1
GUUGGC


UGAGCC





UCAAUG


AACGCA





AAU


CGACG







AD-
CGUGCG
678
3849-3869
AGUUCA
767
3847-3869


571753.1
UUGGCU


UUGAGC





CAAUGA


CAACGC





ACU


ACGAC







AD-
CAAUGA
679
3861-3881
ACGUAG
768
3859-3881


571765.1
ACAGAG


UAUCUC





AUACUA


UGUUCA





CGU


UUGAG







AD-
AAUGAA
680
3862-3882
ACCGUA
769
3860-3882


571766.1
CAGAGA


GUAUCU





UACUAC


CUGUUC





GGU


AUUGA







AD-
AUGAAC
681
3863-3883
AACCGU
770
3861-3883


571767.1
AGAGAU


AGUAUC





ACUACG


UCUGUU





GUU


CAUUG







AD-
CCAAGC
682
3921-3941
AGGUAU
771
3919-3941


571825.1
CUUGGC


UGAGCC





UCAAUA


AAGGCU





CCU


UGGAA







AD-
CAAGCC
683
3922-3942
AUGGUA
772
3920-3942


571826.1
UUGGCU


UUGAGC





CAAUAC


CAAGGC





CAU


UUGGA







AD-
CCACCG
684
4017-4037
AAUUCC
773
4015-4037


571900.1
UAUCCA


CAGUGG





CUGGGA


AUACGG





AUU


UGGGU







AD-
ACCAAG
685
4063-4083
AAAACC
774
4061-4083


571945.1
GAAAAU


CUCAUU





GAGGGU


UUCCUU





UUU


GGUCU







AD-
AAGGAA
686
4066-4086
AGUGAA
775
4064-4086


571948.1
AAUGAG


ACCCUC





GGUUUC


AUUUUC





ACU


CUUGG







AD-
AACUCA
687
4157-4177
AGAAUU
776
4155-4177


572039.1
CCUGUA


UAUUAC





AUAAAU


AGGUGA





UCU


GUUGA







AD-
ACUCAC
688
4158-4178
ACGAAU
777
4156-4178


572040.1
CUGUAA


UUAUUA





UAAAUU


CAGGUG





CGU


AGUUG







AD-
CUCACC
689
4159-4179
AUCGAA
778
4157-4179


572041.1
UGUAAU


UUUAUU





AAAUUC


ACAGGU





GAU


GAGUU







AD-
ACCUGU
690
4162-4182
AAGGUC
779
4160-4182


572044.1
AAUAAA


GAAUUU





UUCGAC


AUUACA





CUU


GGUGA







AD-
UAAUAA
691
4167-4187
ACCUUG
780
4165-4187


572049.1
AUUCGA


AGGUCG





CCUCAA


AAUUUA





GGU


UUACA







AD-
ACCUCA
692
4178-4198
AUUUUA
781
4176-4198


572060.1
AGGUCA


UGGUGA





CCAUAA


CCUUGA





AAU


GGUCG







AD-
CCUCAA
693
4179-4199
AGUUUU
782
4177-4199


572061.1
GGUCAC


AUGGUG





CAUAAA


ACCUUG





ACU


AGGUC







AD-
CUCAAG
694
4180-4200
AGGUUU
783
4178-4200


572062.1
GUCACC


UAUGGU





AUAAAA


GACCUU





CCU


GAGGU







AD-
AGGAUG
695
4226-4246
ACAUAG
784
4224-4246


572108.1
CCAAGA


UGUUCU





ACACUA


UGGCAU





UGU


CCUGA







AD-
CAGAUA
696
4371-4391
ACAUAC
785
4369-4391


572235.1
CAUCUC


UUGGAG





CAAGUA


AUGUAU





UGU


CUGUC







AD-
UGGACA
697
4394-4414
AAUCGG
786
4392-4414


572258.1
AAGCCU


AGAAGG





UCUCCG


CUUUGU





AUU


CCAGC







AD-
AGGAAC
698
4414-4434
AUAGAU
787
4412-4434


572278.1
ACCCUC


GAUGAG





AUCAUC


GGUGUU





UAU


CCUAU







AD-
GGAACA
699
4415-4435
AGUAGA
788
4413-4435


572279.1
CCCUCA


UGAUGA





UCAUCU


GGGUGU





ACU


UCCUA







AD-
AACACC
700
4417-4437
AAGGUA
789
4415-4437


572281.1
CUCAUC


GAUGAU





AUCUAC


GAGGGU





CUU


GUUCC







AD-
CUUUAA
701
4491-4511
AGGAUA
790
4489-4511


572355.1
UGUAGA


AGCUCU





GCUUAU


ACAUUA





CCU


AAGUA







AD-
UUUAAU
702
4492-4512
AUGGAU
791
4490-4512


572356.1
GUAGAG


AAGCUC





CUUAUC


UACAUU





CAU


AAAGU







AD-
UCAAGG
703
4523-4543
AGUAAU
792
4521-4543


572387.1
UCUACG


AGGCGU





CCUAUU


AGACCU





ACU


UGACU







AD-
CAAGGU
704
4524-4544
AUGUAA
793
4522-4544


572388.1
CUACGC


UAGGCG





CUAUUA


UAGACC





CAU


UUGAC







AD-
AAGGUC
705
4525-4545
AUUGUA
794
4523-4545


572389.1
UACGCC


AUAGGC





UAUUAC


GUAGAC





AAU


CUUGA







AD-
AGGUCU
706
4526-4546
AGUUGU
795
4524-4546


572390.1
ACGCCU


AAUAGG





AUUACA


CGUAGA





ACU


CCUUG







AD-
UCUACG
707
4529-4549
ACAGGU
796
4527-4549


572393.1
CCUAUU


UGUAAU





ACAACC


AGGCGU





UGU


AGACC







AD-
AGCUGU
708
4751-4771
AGUCAA
797
4749-477I


572613.1
CCAAUG


AGUCAU





ACUUUG


UGGACA





ACU


GCUGA







AD-
GCUGUC
709
4752-4772
ACGUCA
798
4750-4772


572614.1
CAAUGA


AAGUCA





CUUUGA


UUGGAC





CGU


AGCUG







AD-
AGCAUG
710
5056-5076
ACACCC
799
5054-5076


572858.1
GUUGUC


AAAGAC





UUUGGG


AACCAU





UGU


GCUCU







AD-
AAUAAG
711
1909-1928
UGUCAG
800
1907-1928


890084.1
AAGAAC


UUUGUU





AAACUG


CUUCUU





ACA


AUUCA







AD-
AAUAAG
712
1909-1928
UGUCAG
801
1907-1928


890085.1
AAGAAC


CUUGUU





AAGCUG


CUUCUU





ACA


AUUCA







AD-
AACACC
713
4417-4436
AAGGUA
802
4415-4436


572281
CUCAUC


GAUGAU





AUCUAC


GAGGGU





CUU


GUUCC
















TABLE 7







Modified Sense and Antisense Strand Sequences


of Complement Component C3 dsRNA Agents














Sense
SEQ
Antisense
SEQ
mRNA
SEQ


Duplex
Sequence
ID
Sequence
ID
target 
ID


Name
5′ to 3′
NO:
5′ to 3′
NO:
sequence
NO:
















AD-
asgsac
803
asGfsu
892
CCAGAC
981


568976.1
agAfcA

agAfuG

AGACAA




fAfGfa

fGfucu

GACCAU




ccaucu

uGfuCf

CUACA




acuL96

ugucus








gsg








AD-
ascsag
804
asGfsu
893
AGACAG
982


568978.1
acAfaG

guAfgA

ACAAGA




fAfCfc

fUfggu

CCAUCU




aucuac

cUfuGf

ACACC




acuL96

ucugus








csu








AD-
usgsgg
805
asAfsc
894
CUUGGG
983


569127.1
acAfuU

gaGfuU

ACAUUC




fCfCfg

fCfcgg

CGGAAC




gaacuc

aAfuGf

UCGUC




guuL96

ucccas








asg








AD-
asusuc
806
asAfsu
895
ACAUUC
984


569133.1
cgGfaA

guUfgA

CGGAAC




fCfUfc

fCfgag

UCGUCA




gucaac

uUfcCf

ACAUG




auuL96

ggaaus








gsu








AD-
asgsau
807
asUfsu
896
GAAGAU
985


569164.1
ccGfaG

caUfaG

CCGAGC




fCfCfu

fUfagg

CUACUA




acuaug

cUfcGf

UGAAA




aauL96

gaucus








usc








AD-
gscsag
808
asAfsc
897
CAGCAG
986


569195.1
guCfuU

ucAfgU

GUCUUC




fCfUfc

fGfgag

UCCACU




cacuga

aAfgAf

GAGUU




guuL96

ccugcs








usg








AD-
gscscc
809
asCfsu
898
CUGCCC
987


569237.1
agUfuU

auGfaC

AGUUUC




fCfGfa

fCfucg

GAGGUC




ggucau

aAfaCf

AUAGU




aguL96

ugggcs








asg








AD-
cscsag
810
asCfsa
899
GCCCAG
988


569239.1
uuUfcG

cuAfuG

UUUCGA




fAfGfg

fAfccu

GGUCAU




ucauag

cGfaAf

AGUGG




uguL96

acuggs








gsc








AD-
asasuu
811
asGfsu
900
GAAAUU
989


569272.1
cuAfcU

uaUfaG

CUACUA




fAfCfa

fAfugu

CAUCUA




ucuaua

aGfuAf

UAACG




acuL96

gaauus








usc








AD-
ascsug
812
asCfsc
901
GAACUG
990


569350.1
ccUfuU

gaAfgA

CCUUUG




fGfUfc

fUfgac

UCAUCU




aucuuc

aAfaGf

UCGGG




gguL96

gcagus








usc








AD-
csusca
813
asUfsu
902
ACCUCA
991


569571.1
ugGfuG

cgUfcA

UGGUGU




fUfUfc

fCfgaa

UCGUGA




gugacg

cAfcCf

CGAAC




aauL96

augags








gsu








AD-
usgsgg
814
asGfsu
903
CGUGGG
992


569763.1
caAfcU

aaUfuG

CAACUC




fCfCfa

fUfugg

CAACAA




acaauu

aGfuUf

UUACC




acuL96

gcccas








csg








AD-
gsgsgc
815
asGfsg
904
GUGGGC
993


569764.1
aaCfuC

uaAfuU

AACUCC




fCfAfa

fGfuug

AACAAU




caauua

gAfgUf

UACCU




ccuL96

ugcccs








asc








AD-
gscsaa
816
asCfsa
905
GGGCAA
994


569766.1
cuCfcA

ggUfaA

CUCCAA




fAfCfa

fUfugu

CAAUUA




auuacc

uGfgAf

CCUGC




uguL96

guugcs








csc








AD-
gsusca
817
asAfsu
906
ACGUCA
995


569816.1
acUfuC

ucGfcA

ACUUCC




fCfUfc

fGfgag

UCCUGC




cugcga

gAfaGf

GAAUG




auuL96

uugacs








gsu








AD-
asascu
818
asGfsa
907
CAAACU
996


570156.1
gaCfgC

ucUfuA

GACGCA




fAfGfa

fCfucu

GAGUAA




guaaga

gCfgUf

GAUCU




ucuL96

caguus








usg








AD-
usgsca
819
asAfsa
908
AUUGCA
997


570466.1
gaAfgA

acGfaU

GAAGAG




fGfAfa

fGfuuc

AACAUC




caucgu

uCfuUf

GUUUC




uuuL96

cugcas








asu








AD-
gsasag
820
asCfsg
909
CAGAAG
998


570470.1
agAfaC

ggAfaA

AGAACA




fAfUfc

fCfgau

UCGUUU




guuucc

gUfuCf

CCCGA




cguL96

ucuucs








usg








AD-
asasga
821
asUfsc
910
AGAAGA
999


570471.1
gaAfcA

ggGfaA

GAACAU




fUfCfg

fAfcga

CGUUUC




uuuccc

uGfuUf

CCGAA




gauL96

cucuus








csu








AD-
asgsaa
822
asAfsc
911
AGAGAA
1000


570474.1
caUfcG

uuCfgG

CAUCGU




fUfUfu

fGfaaa

UUCCCG




cccgaa

cGfaUf

AAGUG




guuL96

guucus








csu








AD-
gsasac
823
asCfsa
912
GAGAAC
1001


570475.1
auCfgU

cuUfcG

AUCGUU




fUfUfc

fGfgaa

UCCCGA




ccgaag

aCfgAf

AGUGA




uguL96

uguucs








usc








AD-
asasca
824
asUfsc
913
AGAACA
1002


570476.1
ucGfuU

acUfuC

UCGUUU




fUfCfc

fGfgga

CCCGAA




cgaagu

aAfcGf

GUGAG




gauL96

auguus








csu








AD-
csgsga
825
asAfsc
914
GUCGGA
1003


570620.1
caAfgA

agAfuC

CAAGAA




fAfAfg

fCfcuu

AGGGAU




ggaucu

uCfuUf

CUGUG




guuL96

guccgs








asc








AD-
gsgsac
826
asCfsa
915
UCGGAC
1004


570621.1
aaGfaA

caGfaU

AAGAAA




fAfGfg

fCfccu

GGGAUC




gaucug

uUfcUf

UGUGU




uguL96

uguccs








gsa








AD-
gsasca
827
asAfsc
916
CGGACA
1005


570622.1
agAfaA

acAfgA

AGAAAG




fGfGfg

fUfccc

GGAUCU




aucugu

uUfuCf

GUGUG




guuL96

uugucs








csg








AD-
ascsaa
828
asCfsa
917
GGACAA
1006


570623.1
gaAfaG

caCfaG

GAAAGG




fGfGfa

fAfucc

GAUCUG




ucugug

cUfuUf

UGUGG




uguL96

cuugus








csc








AD-
csasag
829
asCfsc
918
GACAAG
1007


570624.1
aaAfgG

acAfcA

AAAGGG




fGfAfu

fGfauc

AUCUGU




cugugu

cCfuUf

GUGGC




gguL96

ucuugs








usc








AD-
asasga
830
asGfsc
919
ACAAGA
1008


570625.1
aaGfgG

caCfaC

AAGGGA




fAfUfc

fAfgau

UCUGUG




ugugug

cCfcUf

UGGCA




gcuL96

uucuus








gsu








AD-
gsasaa
831
asCfsu
920
AAGAAA
1009


570627.1
ggGfaU

gcCfaC

GGGAUC




fCfUfg

fAfcag

UGUGUG




uguggc

aUfcCf

GCAGA




aguL96

cuuucs








usu








AD-
csusuc
832
asGfsc
921
CCCUUC
1010


570631.1
gaGfgU

auUfaC

GAGGUC




fCfAfc

fUfgug

ACAGUA




aguaau

aCfcUf

AUGCA




gcuL96

cgaags








gsg








AD-
ususcg
833
asUfsg
922
CCUUCG
1011


570632.1
agGfuC

caUfuA

AGGUCA




fAfCfa

fCfugu

CAGUAA




guaaug

gAfcCf

UGCAG




cauL96

ucgaas








gsg








AD-
gsgscu
834
asAfsa
923
GCGGCU
1012


570672.1
acCfcU

caAfcA

ACCCUA




fAfCfu

fGfagu

CUCUGU




cuguug

aGfgGf

UGUUC




uuuL96

uagccs








gsc








AD-
csusac
835
asCfsg
924
GGCUAC
1013


570674.1
ccUfaC

aaCfaA

CCUACU




fUfCfu

fCfaga

CUGUUG




guuguu

gUfaGf

UUCGA




cguL96

gguags








csc








AD-
usascc
836
asUfsc
925
GCUACC
1014


570675.1
cuAfcU

gaAfcA

CUACUC




fCfUfg

fAfcag

UGUUGU




uuguuc

aGfuAf

UCGAA




gauL96

ggguas








gsc








AD-
ascscc
837
asUfsu
926
CUACCC
1015


570676.1
uaCfuC

cgAfaC

UACUCU




fUfGfu

fAfaca

GUUGUU




uguucg

gAfgUf

CGAAA




aauL96

agggus








asg








AD-
cscscu
838
asUfsu
927
UACCCU
1016


570677.1
acUfcU

ucGfaA

ACUCUG




fGfUfu

fCfaac

UUGUUC




guucga

aGfaGf

GAAAC




aauL96

uagggs








usa








AD-
cscsua
839
asGfsu
928
ACCCUA
1017


570678.1
cuCfuG

uuCfgA

CUCUGU




fUfUfg

fAfcaa

UGUUCG




uucgaa

cAfgAf

AAACG




acuL96

guaggs








gsu








AD-
csusac
840
asCfsg
929
CCCUAC
1018


570679.1
ucUfgU

uuUfcG

UCUGUU




fUfGfu

fAfaca

GUUCGA




ucgaaa

aCfaGf

AACGA




cguL96

aguags








gsg








AD-
usascu
841
asUfsc
930
CCUACU
1019


570680.1
cuGfuU

guUfuC

CUGUUG




fGfUfu

fGfaac

UUCGAA




cgaaac

aAfcAf

ACGAG




gauL96

gaguas








gsg








AD-
ascsuc
842
asCfsu
931
CUACUC
1020


570681.1
ugUfuG

cgUfuU

UGUUGU




fUfUfc

fCfgaa

UCGAAA




gaaacg

cAfaCf

CGAGC




aguL96

agagus








asg








AD-
csuscu
843
asGfsc
932
UACUCU
1021


570682.1
guUfgU

ucGfuU

GUUGUU




fUfCfg

fUfcga

CGAAAC




aaacga

aCfaAf

GAGCA




gcuL96

cagags








usa








AD-
cscsgu
844
asCfsc
933
AGCCGU
1022


570717.1
ucUfcU

ggUfaA

UCUCUA




fAfCfa

fUfugu

CAAUUA




auuacc

aGfaGf

CCGGC




gguL96

aacggs








csu








AD-
asasca
845
asCfsg
934
UGAACA
1023


570963.1
aaAfcU

aaCfaG

AAACUG




fGfUfg

fCfcac

UGGCUG




gcuguu

aGfuUf

UUCGC




cguL96

uuguus








csa








AD-
gsgsuc
846
asGfsg
935
ACGGUC
1024


571157.1
auCfgC

uaAfuG

AUCGCU




fUfGfu

fCfaca

GUGCAU




gcauua

gCfgAf

UACCU




ccuL96

ugaccs








gsu








AD-
gsusca
847
asAfsg
936
CGGUCA
1025


571158.1
ucGfcU

guAfaU

UCGCUG




fGfUfg

fGfcac

UGCAUU




cauuac

aGfcGf

ACCUG




cuuL96

augacs








csg








AD-
usgsca
848
asCfsg
937
UGUGCA
1026


571168.1
uuAfcC

uuUfcA

UUACCU




fUfGfg

fUfcca

GGAUGA




augaaa

gGfuAf

AACGG




cguL96

augcas








csa








AD-
csgsug
849
asGfsa
938
UACGUG
1027


571298.1
guCfaA

gaGfaA

GUCAAG




fGfGfu

fGfacc

GUCUUC




cuucuc

uUfgAf

UCUCU




ucuL96

ccacgs








usa








AD-
csgsgc
850
asCfsg
939
CACGGC
1028


571447.1
cuUfuG

agAfuG

CUUUGU




fUfUfc

fAfgaa

UCUCAU




ucaucu

cAfaAf

CUCGC




cguL96

ggccgs








usg








AD-
gsgscc
851
asGfsc
940
ACGGCC
1029


571448.1
uuUfgU

gaGfaU

UUUGUU




fUfCfu

fGfaga

CUCAUC




caucuc

aCfaAf

UCGCU




gcuL96

aggccs








gsu








AD-
gscscu
852
asAfsg
941
CGGCCU
1030


571449.1
uuGfuU

cgAfgA

UUGUUC




fCfUfc

fUfgag

UCAUCU




aucucg

aAfcAf

CGCUG




cuuL96

aaggcs








csg








AD-
ususcc
853
asAfsu
942
ACUUCC
1031


571539.1
uuGfaA

guAfgU

UUGAAG




fGfCfc

fUfggc

CCAACU




aacuac

uUfcAf

ACAUG




auuL96

aggaas








gsu








AD-
usgsca
854
asGfsu
943
ACUGCA
1032


571719.1
gcUfaA

caAfaG

GCUAAA




fAfAfg

fUfcuu

AGACUU




acuuug

uUfaGf

UGACU




acuL96

cugcas








gsu








AD-
uscsgu
855
asUfsu
944
CGUCGU
1033


571752.1
gcGfuU

caUfuG

GCGUUG




fGfGfc

fAfgcc

GCUCAA




ucaaug

aAfcGf

UGAAC




aauL96

cacgas








csg








AD-
csgsug
856
asGfsu
945
GUCGUG
1034


571753.1
cgUfuG

ucAfuU

CGUUGG




fGfCfu

fGfagc

CUCAAU




caauga

cAfaCf

GAACA




acuL96

gcacgs








asc








AD-
csasau
857
asCfsg
946
CUCAAU
1035


571765.1
gaAfcA

uaGfuA

GAACAG




fGfAfg

fUfcuc

AGAUAC




auacua

uGfuUf

UACGG




cguL96

cauugs








asg








AD-
asasug
858
asCfsc
947
UCAAUG
1036


571766.1
aaCfaG

guAfgU

AACAGA




fAfGfa

fAfucu

GAUACU




uacuac

cUfgUf

ACGGU




gguL96

ucauus








gsa








AD-
asusga
859
asAfsc
948
CAAUGA
1037


571767.1
acAfgA

cgUfaG

ACAGAG




fGfAfu

fUfauc

AUACUA




acuacg

uCfuGf

CGGUG




guuL96

uucaus








usg








AD-
cscsaa
860
asGfsg
949
UUCCAA
1038


571825.1
gcCfuU

uaUfuG

GCCUUG




fGfGfc

fAfgcc

GCUCAA




ucaaua

aAfgGf

UACCA




ccuL96

cuuggs








asa








AD-
csasag
861
asUfsg
950
UCCAAG
1039


571826.1
ccUfuG

guAfuU

CCUUGG




fGfCfu

fGfagc

CUCAAU




caauac

cAfaGf

ACCAA




cauL96

gcuugs








gsa








AD-
cscsac
862
asAfsu
951
ACCCAC
1040


571900.1
cgUfaU

ucCfcA

CGUAUC




fCfCfa

fGfugg

CACUGG




cuggga

aUfaCf

GAAUC




auuL96

gguggs








gsu








AD-
ascsca
863
asAfsa
952
AGACCA
1041


571945.1
agGfaA

acCfcU

AGGAAA




fAfAfu

fCfauu

AUGAGG




gagggu

uUfcCf

GUUUC




uuuL96

uuggus








csu








AD-
asasgg
864
asGfsu
953
CCAAGG
1042


571948.1
aaAfaU

gaAfaC

AAAAUG




fGfAfg

fCfcuc

AGGGUU




gguuuc

aUfuUf

UCACA




acuL96

uccuus








gsg








AD-
asascu
865
asGfsa
954
UCAACU
1043


572039.1
caCfcU

auUfuA

CACCUG




fGfUfa

fUfuac

UAAUAA




auaaau

aGfgUf

AUUCG




ucuL96

gaguus








gsa








AD-
ascsuc
866
asCfsg
955
CAACUC
1044


572040.1
acCfuG

aaUfuU

ACCUGU




fUfAfa

fAfuua

AAUAAA




uaaauu

cAfgGf

UUCGA




cguL96

ugagus








usg








AD-
csusca
867
asUfsc
956
AACUCA
1045


572041.1
ccUfgU

gaAfuU

CCUGUA




fAfAfu

fUfauu

AUAAAU




aaauuc

aCfaGf

UCGAC




gauL96

gugags








usu








AD-
ascscu
868
asAfsg
957
UCACCU
1046


572044.1
guAfaU

guCfgA

GUAAUA




fAfAfa

fAfuuu

AAUUCG




uucgac

aUfuAf

ACCUC




cuuL96

caggus








gsa








AD-
usasau
869
asCfsc
958
UGUAAU
1047


572049.1
aaAfuU

uuGfaG

AAAUUC




fCfGfa

fGfucg

GACCUC




ccucaa

aAfuUf

AAGGU




gguL96

uauuas








csa








AD-
ascscu
870
asUfsu
959
CGACCU
1048


572060.1
caAfgG

uuAfuG

CAAGGU




fUfCfa

fGfuga

CACCAU




ccauaa

cCfuUf

AAAAC




aauL96

gaggus








csg








AD-
cscsuc
871
asGfsu
960
GACCUC
1049


572061.1
aaGfgU

uuUfaU

AAGGUC




fCfAfc

fGfgug

ACCAUA




cauaaa

aCfcUf

AAACC




acuL96

ugaggs








usc








AD-
csusca
872
asGfsg
961
ACCUCA
1050


572062.1
agGfuC

uuUfuA

AGGUCA




fAfCfc

fUfggu

CCAUAA




auaaaa

gAfcCf

AACCA




ccuL96

uugags








gsu








AD-
asgsga
873
asCfsa
962
UCAGGA
1051


572108.1
ugCfcA

uaGfuG

UGCCAA




fAfGfa

fUfucu

GAACAC




acacua

uGfgCf

UAUGA




uguL96

auccus








gsa








AD-
csasga
874
asCfsa
963
GACAGA
1052


572235.1
uaCfaU

uaCfuU

UACAUC




fCfUfc

fGfgag

UCCAAG




caagua

aUfgUf

UAUGA




uguL96

aucugs








usc








AD-
usgsga
875
asAfsu
964
GCUGGA
1053


572258.1
caAfaG

cgGfaG

CAAAGC




fCfCfu

fAfagg

CUUCUC




ucuccg

cUfuUf

CGAUA




auuL96

guccas








gsc








AD-
asgsga
876
asUfsa
965
AUAGGA
1054


572278.1
acAfcC

gaUfgA

ACACCC




fCfUfc

fUfgag

UCAUCA




aucauc

gGfuGf

UCUAC




uauL96

uuccus








asu








AD-
gsgsaa
877
asGfsu
966
UAGGAA
1055


572279.1
caCfcC

agAfuG

CACCCU




fUfCfa

fAfuga

CAUCAU




ucaucu

gGfgUf

CUACC




acuL96

guuccs








usa








AD-
asasca
878
asAfsg
967
GGAACA
1056


572281.1
ccCfuC

guAfgA

CCCUCA




fAfUfc

fUfgau

UCAUCU




aucuac

gAfgGf

ACCUG




cuuL96

guguus








csc








AD-
csusuu
879
asGfsg
968
UACUUU
1057


572355.1
aaUfgU

auAfaG

AAUGUA




fAfGfa

fCfucu

GAGCUU




gcuuau

aCfaUf

AUCCA




ccuL96

uaaags








usa








AD-
ususua
880
asUfsg
969
ACUUUA
1058


572356.1
auGfuA

gaUfaA

AUGUAG




fGfAfg

fGfcuc

AGCUUA




cuuauc

uAfcAf

UCCAG




cauL96

uuaaas








gsu








AD-
uscsaa
881
asGfsu
970
AGUCAA
1059


572387.1
ggUfcU

aaUfaG

GGUCUA




fAfCfg

fGfcgu

CGCCUA




ccuauu

aGfaCf

UUACA




acuL96

cuugas








csu








AD-
csasag
882
asUfsg
971
GUCAAG
1060


572388.1
guCfuA

uaAfuA

GUCUAC




fCfGfc

fGfgcg

GCCUAU




cuauua

uAfgAf

UACAA




cauL96

ccuugs








asc








AD-
asasgg
883
asUfsu
972
UCAAGG
1061


572389.1
ucUfaC

guAfaU

UCUACG




fGfCfc

fAfggc

CCUAUU




uauuac

gUfaGf

ACAAC




aauL96

accuus








gsa








AD-
asgsgu
884
asGfsu
973
CAAGGU
1062


572390.1
cuAfcG

ugUfaA

CUACGC




fCfCfu

fUfagg

CUAUUA




auuaca

cGfuAf

CAACC




acuL96

gaccus








usg








AD-
uscsua
885
asCfsa
974
GGUCUA
1063


572393.1
cgCfcU

ggUfuG

CGCCUA




fAfUfu

fUfaau

UUACAA




acaacc

aGfgCf

CCUGG




uguL96

guagas








csc








AD-
asgscu
886
asGfsu
975
UCAGCU
1064


572613.1
guCfcA

caAfaG

GUCCAA




fAfUfg

fUfcau

UGACUU




acuuug

uGfgAf

UGACG




acuL96

cagcus








gsa








AD-
gscsug
887
asCfsg
976
CAGCUG
1065


572614.1
ucCfaA

ucAfaA

UCCAAU




fUfGfa

fGfuca

GACUUU




cuuuga

uUfgGf

GACGA




cguL96

acagcs








usg








AD-
asgsca
888
asCfsa
977
AGAGCA
1066


572858.1
ugGfuU

ccCfaA

UGGUUG




fGfUfc

fAfgac

UCUUUG




uuuggg

aAfcCf

GGUGC




uguL96

augcus








csu








AD-
asasua
889
usGfsu
978
AAUAAG
1067


890084.1
agAfaG

caGfuu

AAGAAC




fAfAfc

uguucU

AAACUG




aaacug

fuCfuu

ACA




acaL96

auuscs








a








AD-
asasua
890
usGfsu
979
AAUAAG
1068


890085.1
agAfaG

caGfcu

AAGAAC




fAfAfc

uguucU

AAGCUG




aagcug

fuCfuu

ACA




acaL96

auuscs








a








AD-
asasca
891
asAfsg
980
AACACC
1069


572281.1
ccCfuC

guAfgA

CUCAUC




fAfUfc

fUfgau

AUCUAC




aucuac

gAfgGf

CUU




cuuL96

guguus








csc



















TABLE 8







C3 Single Dose Screens in Hep3B cells











10 nM Dose
1.0 nM Dose
0.1 nM Dose














Avg % C3

Avg % C3

Avg % C3




mRNA

mRNA

mRNA



Duplex
Remaining
SD
Remaining
SD
Remaining
SD
















AD-565279.1
17.6
6.0
54.0
11.4
99.8
17.7


AD-565541.1
7.7
2.3
23.4
2.0
72.7
10.0


AD-566234.1
32.9
4.8
66.9
7.3
98.1
21.5


AD-566383.1
36.8
7.3
66.7
2.6
105.4
21.7


AD-566412.1
15.9
5.1
43.0
2.8
94.9
27.3


AD-566444.1
12.6
0.9
50.3
5.1
88.9
13.5


AD-566448.1
25.4
9.3
43.0
6.7
107.7
18.3


AD-567066.1
10.0
2.4
44.9
3.5
91.0
28.2


AD-567307.1
21.5
2.9
48.6
8.7
94.2
10.7


AD-567487.1
21.0
6.6
49.0
5.6
67.6
23.5


AD-567700.1
12.9
1.8
39.5
4.4
95.0
18.1


AD-567716.1
27.5
6.9
59.0
8.0
110.5
30.4


AD-568003.1
18.5
3.7
73.3
4.6
113.3
13.5


AD-568026.1
11.8
1.4
32.5
7.5
51.7
10.5


AD-568157.1
22.5
6.4
40.0
5.7
80.6
15.0


AD-568586.1
9.9
1.2
28.0
5.1
91.5
9.8


AD-566445.1
22.4
8.4
60.0
1.8
108.5
15.0


AD-567812.1
35.2
8.2
60.2
8.6
100.7
16.7


AD-564901.1
57.2
9.3
95.2
7.4
100.3
29.6


AD-566446.1
55.5
3.2
96.6
8.4
114.1
8.2


AD-566409.1
80.5
30.3
63.1
36.1
95.5
12.2


AD-567067.1
21.5
15.5
44.3
5.7
101.3
12.9


AD-568160.1
18.5
1.5
49.1
9.3
72.5
16.8


AD-565282.1
27.3
1.6
51.0
7.1
102.9
20.4


AD-568344.1
33.7
6.9
85.5
4.1
121.9
23.9


AD-567304.1
9.9
2.3
22.2
3.8
64.8
6.1


AD-568153.1
24.3
3.5
53.8
7.0
100.9
12.3


AD-564742.1
8.7
1.4
20.6
7.9
63.6
21.3


AD-565284.1
13.6
3.5
45.4
6.4
102.5
16.7


AD-566485.1
96.5
15.4
112.4
7.6
110.9
13.7


AD-567808.1
65.1
14.3
94.5
6.0
118.2
14.7


AD-566449.1
89.3
5.8
117.1
7.1
114.3
17.4


AD-568382.1
50.5
10.8
98.3
2.5
125.4
13.6


AD-566442.1
36.4
5.2
94.3
9.5
116.4
16.5


AD-567809.1
81.5
23.1
93.8
8.1
121.5
19.1


AD-565534.1
106.1
24.3
109.8
7.0
113.5
8.7


AD-567215.1
55.0
6.0
95.8
3.1
94.2
13.4


AD-566443.1
79.9
7.3
117.1
7.8
126.2
9.8


AD-568156.1
54.1
6.9
76.5
5.9
72.0
8.4


AD-565532.1
65.0
13.4
101.8
3.6
105.2
22.1


AD-566447.1
50.4
5.4
98.9
5.4
125.3
9.0


AD-565040.1
99.7
12.0
111.2
7.0
124.6
11.5


AD-568161.1
59.7
8.4
86.9
12.5
82.7
8.9


AD-567829.1
57.9
10.9
96.9
12.3
102.6
22.1


AD-564975.1
106.6
9.3
102.3
25.2
126.7
10.1


AD-567713.1
10.8
3.5
23.4
1.6
70.1
21.0


AD-566411.1
32.5
3.8
65.2
8.7
112.7
41.1


AD-565005.1
42.2
5.8
84.7
12.3
99.7
15.5


AD-567156.1
44.6
21.2
75.5
20.7
94.1
21.1


AD-566388.1
66.6
8.2
105.7
6.4
99.6
10.6


AD-566384.1
32.2
5.9
75.8
7.0
115.8
19.0


AD-564744.1
65.3
14.4
96.4
5.8
122.2
34.5


AD-567828.1
99.9
6.9
108.7
7.4
113.4
14.1


AD-567063.1
33.0
11.0
67.4
6.6
92.1
20.3


AD-568158.1
74.1
8.0
85.4
9.6
87.1
10.8


AD-567521.1
12.7
5.8
24.5
5.6
70.6
9.1


AD-567395.1
78.7
14.6
101.5
8.5
106.0
18.7


AD-567582.1
65.5
9.4
82.3
4.5
112.3
17.2


AD-564745.1
20.0
5.7
61.2
7.6
105.8
21.7


AD-567831.1
68.7
11.4
100.1
7.2
123.3
16.4


AD-565535.1
60.1
9.4
86.7
11.8
103.8
20.5


AD-564730.1
14.3
6.9
41.4
3.4
95.1
6.2


AD-567318.1
25.4
2.1
69.7
6.3
107.0
17.4


AD-567314.1
101.9
4.2
115.5
8.4
103.6
22.1


AD-568341.1
67.0
18.2
92.7
11.0
104.7
20.5


AD-568099.1
14.5
3.6
60.6
7.4
113.5
13.5


AD-566837.1
7.1
1.5
31.7
4.8
88.3
22.0


AD-565616.1
95.7
9.4
95.0
20.1
122.7
14.1


AD-568345.1
40.4
5.4
83.9
8.7
114.5
14.7


AD-565925.1
27.8
6.2
70.1
3.5
103.8
13.3


AD-564727.1
25.2
5.1
78.1
9.4
103.6
19.1


AD-565281.1
24.9
3.8
54.3
13.4
88.2
15.4


AD-565278.1
20.5
2.6
66.5
15.7
106.0
27.8


AD-564976.1
80.3
4.1
96.9
8.9
90.8
13.7


AD-568343.1
20.4
5.5
35.1
20.5
79.8
7.7


AD-568100.1
11.5
2.6
35.5
4.5
81.4
12.5


AD-566935.1
42.0
8.9
80.6
7.9
116.6
12.1


AD-567315.1
7.5
0.9
11.0
1.7
43.7
12.0


AD-566386.1
25.0
2.8
62.7
13.1
94.7
18.4


AD-567813.1
27.6
2.6
61.8
5.8
118.2
20.3


AD-564739.1
46.7
11.9
66.3
3.8
117.7
28.3


AD-564731.1
56.7
15.0
95.2
3.8
117.1
21.3


AD-565904.1
7.8
4.4
23.6
4.3
64.5
16.1


AD-566528.1
32.0
7.8
64.3
12.3
102.7
27.8


AD-567699.1
86.1
8.4
104.8
6.3
116.5
16.2


AD-565905.1
33.3
19.4
58.9
5.2
96.9
12.4


AD-567814.1
11.3
2.0
30.8
5.1
95.8
20.8


AD-568381.1
87.3
15.7
92.7
8.5
117.0
10.6
















TABLE 9







C3 Single Dose Screens in PMH cells











10 nM Dose
1.0 nM Dose
0.1 nM Dose














Avg % C3

Avg % C3

Avg % C3




mRNA

mRNA

mRNA



Duplex
Remaining
SD
Remaining
SD
Remaining
SD
















AD-565279.1
31.0
8.3
57.8
10.4
123.1
8.8


AD-565541.1
110.9
7.6
108.5
2.9
97.3
23.7


AD-566234.1
94.2
8.9
77.0
35.4
105.4
9.0


AD-566383.1
89.7
24.3
54.8
31.0
68.6
39.1


AD-566412.1
30.0
4.1
38.3
14.5
88.1
17.0


AD-566444.1
110.6
12.5
102.6
6.7
105.7
48.0


AD-566448.1
127.1
8.0
84.0
14.7
120.8
9.1


AD-567066.1
21.4
5.9
33.1
8.0
100.5
24.8


AD-567307.1
110.7
9.0
111.3
5.7
84.8
43.9


AD-567487.1
105.8
12.4
77.2
7.4
100.9
15.5


AD-567700.1
22.6
4.5
44.4
3.7
68.7
25.3


AD-567716.1
122.0
6.3
102.5
4.2
93.1
15.6


AD-568003.1
110.4
22.4
104.7
4.6
115.9
20.6


AD-568026.1
55.1
16.9
81.5
7.8
94.4
7.1


AD-568157.1
60.9
8.8
83.2
9.9
102.2
36.7


AD-568586.1
106.4
11.9
105.0
8.4
103.7
24.9


AD-566445.1
110.3
4.8
90.3
9.2
104.2
12.0


AD-567812.1
111.8
7.1
91.8
8.4
127.5
11.1


AD-564901.1
120.0
8.1
109.3
8.0
104.2
20.7


AD-566446.1
112.7
16.7
92.6
10.3
100.9
19.3


AD-566409.1
109.1
18.7
52.0
17.7
90.7
21.6


AD-567067.1
15.7
3.2
22.5
8.9
80.9
30.8


AD-568160.1
87.2
8.0
97.5
7.6
92.3
19.4


AD-565282.1
30.4
8.7
63.1
3.0
99.9
9.2


AD-568344.1
36.7
4.9
77.8
13.7
104.8
8.8


AD-567304.1
88.4
16.6
100.0
15.1
81.0
48.3


AD-568153.1
87.3
3.7
100.4
8.2
97.9
34.6


AD-564742.1
20.2
1.6
34.7
8.4
67.3
14.8


AD-565284.1
25.1
4.2
48.7
3.9
103.5
29.2


AD-566485.1
93.8
28.0
113.5
8.5
96.8
23.3


AD-567808.1
112.5
18.1
86.2
6.1
98.7
10.8


AD-566449.1
123.5
9.0
81.7
27.6
96.6
40.4


AD-568382.1
111.9
17.7
107.5
9.6
107.5
13.9


AD-566442.1
109.7
9.7
100.0
6.9
105.7
20.8


AD-567809.1
97.6
13.6
54.0
29.7
117.1
5.6


AD-565534.1
114.9
6.8
113.2
5.9
110.6
8.6


AD-567215.1
105.5
19.2
85.6
12.3
111.1
3.6


AD-566443.1
119.7
12.3
109.3
5.2
109.5
24.2


AD-568156.1
72.9
9.7
91.2
4.7
97.7
9.5


AD-565532.1
102.4
10.2
103.5
6.8
98.0
36.3


AD-566447.1
114.2
4.4
102.7
4.5
88.7
37.2


AD-565040.1
127.8
15.7
98.6
11.2
104.0
7.1


AD-568161.1
88.2
10.4
93.5
9.8
98.4
9.7


AD-567829.1
108.9
9.4
76.7
10.5
132.9
16.7


AD-564975.1
118.7
12.2
97.5
7.0
110.0
23.1


AD-567713.1
111.7
11.6
97.8
12.0
64.6
36.6


AD-566411.1
76.4
10.7
63.1
18.3
98.9
20.4


AD-565005.1
113.6
7.4
111.2
8.5
76.2
23.3


AD-567156.1
78.3
16.8
63.4
6.6
73.5
22.2


AD-566388.1
80.3
12.0
83.6
14.1
109.9
12.6


AD-566384.1
76.2
10.0
79.3
12.7
120.1
15.7


AD-564744.1
38.1
10.0
63.6
24.1
91.0
39.1


AD-567828.1
100.8
23.0
91.7
13.7
108.9
24.8


AD-567063.1
27.0
7.4
33.6
14.4
97.3
18.1


AD-568158.1
87.9
13.0
116.6
9.6
108.7
18.3


AD-567521.1
74.5
12.0
93.9
5.5
95.0
32.1


AD-567395.1
87.6
6.2
78.3
12.1
118.8
7.3


AD-567582.1
85.3
11.3
83.0
7.8
105.4
20.4


AD-564745.1
24.6
1.7
45.6
3.0
101.2
22.3


AD-567831.1
112.4
7.6
106.1
12.3
93.4
32.9


AD-565535.1
85.8
13.2
97.5
12.1
96.6
39.6


AD-564730.1
21.1
3.0
29.7
14.9
98.8
9.6


AD-567318.1
56.0
11.2
93.9
4.2
125.9
12.0


AD-567314.1
119.1
12.5
105.2
7.7
99.9
34.0


AD-568341.1
126.3
18.8
82.3
26.2
97.9
28.5


AD-568099.1
133.5
18.4
102.6
1.3
110.5
7.7


AD-566837.1
42.1
11.7
55.3
7.2
108.7
18.8


AD-565616.1
38.7
7.6
59.5
5.7
99.1
13.6


AD-568345.1
38.7
8.2
66.4
7.6
101.7
5.9


AD-565925.1
117.3
12.9
106.3
3.0
92.6
41.3


AD-564727.1
37.2
7.4
59.3
4.3
95.8
11.0


AD-565281.1
18.8
3.6
25.2
13.0
47.7
30.8


AD-565278.1
61.2
11.1
77.2
8.0
91.3
44.1


AD-564976.1
76.0
25.2
29.2
6.5
71.5
27.3


AD-568343.1
29.3
3.3
30.2
8.5
83.6
19.4


AD-568100.1
109.2
23.2
86.6
14.7
117.8
12.8


AD-566935.1
128.7
12.5
98.3
9.8
85.2
37.1


AD-567315.1
47.0
3.5
78.1
11.0
110.6
9.0


AD-566386.1
65.3
17.0
64.6
11.3
132.8
17.4


AD-567813.1
111.8
19.7
99.0
9.8
79.9
24.2


AD-564739.1
21.0
2.2
46.8
3.1
112.7
8.6


AD-564731.1
71.0
11.9
67.2
26.6
83.0
41.0


AD-565904.1
65.5
14.7
60.6
23.4
92.2
17.5


AD-566528.1
97.2
16.5
114.1
10.6
103.9
19.7


AD-567699.1
117.3
17.7
74.6
17.0
76.0
38.7


AD-565905.1
89.0
13.7
74.0
15.0
92.0
11.1


AD-567814.1
106.6
24.8
103.0
3.8
123.8
4.9


AD-568381.1
112.1
5.9
104.6
7.4
84.8
25.0
















TABLE 10







C3 Single Dose Screens in Hep3B cells











10 nM Dose
1.0 nM Dose
0.1 nM Dose














Avg % C3

Avg % C3

Avg % C3




mRNA

mRNA

mRNA



Duplex
Remaining
SD
Remaining
SD
Remaining
SD
















AD-569034.1
17.5
3.2
50.3
12.4
81.5
20.9


AD-569164.1
9.7
1.6
22.3
2.7
43.9
7.0


AD-569165.1
20.8
1.8
51.1
9.3
80.0
15.2


AD-569272.1
14.2
0.3
44.0
9.9
78.5
9.2


AD-569763.1
9.6
1.2
41.8
4.9
74.9
5.6


AD-569765.1
13.4
2.2
41.7
9.5
83.1
29.8


AD-570130.1
10.8
0.9
27.6
9.0
49.1
6.3


AD-570132.1
18.0
3.3
57.7
2.8
59.3
5.8


AD-570133.1
23.9
4.8
70.8
13.0
114.2
19.8


AD-570134.1
9.3
4.3
18.1
4.6
31.1
5.5


AD-570157.1
14.7
1.2
50.1
13.8
92.4
13.6


AD-570711.1
11.3
1.1
33.5
5.1
70.8
9.0


AD-570712.1
7.6
1.0
20.2
2.2
51.0
11.2


AD-570713.1
8.5
2.5
13.5
2.4
37.6
11.3


AD-570714.1
7.5
2.2
16.2
5.2
35.3
7.4


AD-571539.1
4.6
0.1
18.5
2.9
28.4
4.7


AD-571610.1
12.5
2.3
41.2
6.8
77.5
11.3


AD-571633.1
20.2
2.5
65.1
12.8
73.6
5.0


AD-571715.1
6.1
1.0
18.2
5.8
46.0
7.8


AD-571752.1
8.7
1.8
20.2
3.3
51.7
12.8


AD-571754.1
23.1
2.4
67.0
12.4
97.1
28.4


AD-571828.1
28.9
2.9
61.6
11.2
84.0
8.4


AD-572039.1
16.6
3.1
46.0
13.7
83.5
12.2


AD-572040.1
10.3
2.6
28.4
4.8
67.1
21.6


AD-572041.1
16.0
1.8
42.3
14.6
76.0
21.7


AD-572059.1
12.9
2.8
36.9
7.1
77.2
14.1


AD-572061.1
17.2
5.1
39.2
6.0
74.3
19.6


AD-572062.1
11.6
2.2
31.0
1.7
63.4
10.0


AD-572063.1
14.5
1.2
41.7
5.8
81.0
15.5


AD-572110.1
10.4
1.1
25.5
6.6
63.3
18.8


AD-572144.1
13.3
1.6
41.7
3.4
94.6
10.9


AD-572388.1
12.8
2.1
33.3
4.1
63.8
19.8


AD-572389.1
9.8
1.5
13.6
1.8
32.1
7.5


AD-572390.1
14.2
1.6
38.7
6.8
74.2
7.7
















TABLE 11







C3 Single Dose Screens in PMH cells











10 nM Dose
1.0 nM Dose
0.1 nM Dose














Avg % C3

Avg % C3

Avg % C3




mRNA

mRNA

mRNA



Duplex
Remaining
SD
Remaining
SD
Remaining
SD
















AD-569034.1
87.3
9.8
94.4
8.2
83.5
8.5


AD-569164.1
66.6
3.9
85.1
21.3
77.2
4.3


AD-569165.1
86.3
12.7
106.1
12.8
101.9
9.8


AD-569272.1
92.1
13.5
89.2
21.7
91.8
7.6


AD-569763.1
42.3
10.4
93.0
16.4
100.8
13.7


AD-569765.1
28.7
2.9
64.2
4.6
97.3
7.6


AD-570130.1
23.8
3.5
68.5
14.7
81.8
11.3


AD-570132.1
72.5
11.6
86.6
9.6
76.3
9.4


AD-570133.1
76.6
15.4
86.6
22.3
80.1
10.7


AD-570134.1
9.6
1.4
24.8
5.4
66.3
7.5


AD-570157.1
92.0
12.3
108.1
7.4
95.7
5.4


AD-570711.1
90.0
25.1
84.6
14.0
104.5
21.7


AD-570712.1
102.1
7.5
95.6
12.2
97.8
12.7


AD-570713.1
99.4
4.9
110.2
8.0
94.0
18.2


AD-570714.1
87.7
2.9
113.2
9.6
87.0
11.6


AD-571539.1
60.2
14.0
84.8
18.0
78.9
11.1


AD-571610.1
83.0
16.3
96.7
4.4
88.4
8.4


AD-571633.1
66.6
15.3
70.6
17.2
87.1
14.4


AD-571715.1
16.0
2.9
50.2
4.4
90.6
8.1


AD-571752.1
94.9
5.4
99.5
10.1
111.4
12.4


AD-571754.1
96.0
5.4
90.2
18.5
103.7
9.1


AD-571828.1
61.1
8.9
98.2
4.9
100.1
5.6


AD-572039.1
99.8
5.3
110.7
22.2
91.1
13.8


AD-572040.1
97.2
10.0
104.4
22.2
81.8
20.1


AD-572041.1
93.3
15.6
81.2
19.7
90.5
11.0


AD-572059.1
101.3
15.9
88.7
14.1
105.2
15.1


AD-572061.1
101.0
6.6
74.1
18.2
113.5
11.3


AD-572062.1
80.4
14.4
102.8
18.6
101.3
10.4


AD-572063.1
100.9
7.7
90.7
22.2
113.7
15.3


AD-572110.1
91.4
10.4
98.0
14.6
108.1
9.9


AD-572144.1
102.7
7.4
90.0
32.4
108.5
10.8


AD-572388.1
17.9
2.8
48.6
3.7
85.3
6.9


AD-572389.1
8.7
2.9
27.6
7.1
73.7
8.1


AD-572390.1
26.8
6.0
60.1
13.2
102.5
4.4
















TABLE 12







C3 Single Dose Screens in Hep3B cells













Avg % C3






mRNA

Dose



Duplex
Remaining
SD
nM















AD-568976.1
14.7
0.2
50



AD-568978.1
14.2
4.1
50



AD-569127.1
16.7
1.8
50



AD-569133.1
21.6
1.3
50



AD-569164.3
21.4
5.8
50



AD-569164.4
22.1
3.3
50



AD-569195.1
22.7
6.8
50



AD-569237.1
103.6
5.6
50



AD-569239.1
76.5
2.8
50



AD-569272.3
26.3
2.2
50



AD-569350.1
63.8
6.4
50



AD-569571.1
19.1
7.6
50



AD-569763.3
20.9
3.5
50



AD-569764.1
18.7
2.1
50



AD-569766.1
74.4
21.6
50



AD-569816.1
21.0
5.5
50



AD-570156.1
19.2
2.5
50



AD-570466.1
23.1
1.4
50



AD-570470.1
59.1
10.2
50



AD-570471.1
36.8
8.3
50



AD-570474.1
54.0
8.4
50



AD-570475.1
35.7
4.9
50



AD-570476.1
22.4
6.3
50



AD-570620.1
16.1
2.5
50



AD-570621.1
20.8
3.7
50



AD-570622.1
16.1
5.7
50



AD-570623.1
16.7
2.8
50



AD-570624.1
20.6
1.5
50



AD-570625.1
19.5
5.5
50



AD-570627.1
20.5
4.1
50



AD-570631.1
26.5
3.0
50



AD-570632.1
24.7
5.2
50



AD-570672.1
21.2
4.7
50



AD-570674.1
33.5
15.3
50



AD-570675.1
107.8
1.7
50



AD-570676.1
64.7
13.8
50



AD-570677.1
29.9
3.0
50



AD-570678.1
102.7
3.7
50



AD-570679.1
49.1
6.4
50



AD-570680.1
50.0
8.0
50



AD-570681.1
23.6
4.2
50



AD-570682.1
27.5
3.8
50



AD-570717.1
83.2
11.9
50



AD-570963.1
28.9
6.5
50



AD-571157.1
61.5
5.2
50



AD-571158.1
96.6
6.2
50



AD-571168.1
62.8
7.7
50



AD-571298.1
12.9
2.7
50



AD-571298.2
9.0
1.6
50



AD-571447.1
49.9
2.1
50



AD-571448.1
28.3
7.7
50



AD-571449.1
78.7
11.7
50



AD-571539.4
21.9
4.8
50



AD-571719.1
14.9
2.7
50



AD-571752.3
29.0
2.4
50



AD-571753.1
19.0
3.9
50



AD-571765.1
41.6
11.4
50



AD-571766.1
25.1
4.4
50



AD-571767.1
23.8
1.0
50



AD-571825.1
15.1
0.9
50



AD-571826.1
17.3
1.3
50



AD-571900.1
25.1
2.6
50



AD-571945.1
23.6
8.1
50



AD-571948.1
89.7
19.3
50



AD-572039.3
34.2
13.5
50



AD-572040.3
26.6
3.7
50



AD-572041.3
25.6
0.6
50



AD-572044.1
25.4
5.4
50



AD-572049.1
31.9
4.3
50



AD-572060.1
25.5
4.8
50



AD-572061.2
24.8
8.1
50



AD-572062.3
23.3
4.5
50



AD-572108.1
61.8
0.9
50



AD-572235.1
17.7
3.1
50



AD-572258.1
14.9
3.3
50



AD-572278.1
14.7
5.5
50



AD-572279.1
14.6
2.1
50



AD-572281.1
13.9
1.3
50



AD-572355.1
70.2
6.2
50



AD-572356.1
22.5
5.7
50



AD-57238.2
15.7
5.3
50



AD-572387.1
15.3
0.5
50



AD-572388.4
14.8
2.3
50



AD-572389.3
12.1
1.1
50



AD-572390.2
15.0
4.1
50



AD-572393.1
19.6
2.3
50



AD-572613.1
125.8
13.8
50



AD-572614.1
30.9
4.9
50



AD-572858.1
26.7
4.0
50



AD-80806.9
11.9
2.4
50



AD-890084.1
15.9
2.2
50



AD-890085.1
43.1
2.4
50



AD-568976.1
25.2
4.3
10



AD-568978.1
18.0
0.5
10



AD-569127.1
22.6
8.6
10



AD-569133.1
33.5
10.9
10



AD-569164.3
10.7
0.6
10



AD-569164.4
37.7
15.6
10



AD-569195.1
18.3
2.3
10



AD-569237.1
106.9
2.2
10



AD-569239.1
123.8
22.4
10



AD-569272.3
63.0
8.3
10



AD-569350.1
106.7
8.7
10



AD-569571.1
21.3
0.8
10



AD-569763.3
34.1
5.6
10



AD-569764.1
37.9
4.5
10



AD-569766.1
94.8
14.9
10



AD-569816.1
13.7
0.9
10



AD-570156.1
27.0
2.4
10



AD-570466.1
14.7
2.7
10



AD-570470.1
95.6
16.6
10



AD-570471.1
48.4
5.1
10



AD-570474.1
25.6
2.9
10



AD-570475.1
84.4
20.6
10



AD-570476.1
26.7
5.9
10



AD-570620.1
22.3
0.5
10



AD-570621.1
31.7
9.8
10



AD-570622.1
10.9
2.1
10



AD-570623.1
22.5
3.4
10



AD-570624.1
25.2
2.6
10



AD-570625.1
14.8
0.3
10



AD-570627.1
29.9
3.2
10



AD-570631.1
35.9
4.2
10



AD-570632.1
38.5
2.2
10



AD-570672.1
39.4
6.6
10



AD-570674.1
34.3
4.0
10



AD-570675.1
97.7
8.2
10



AD-570676.1
86.9
3.8
10



AD-570677.1
60.3
1.4
10



AD-570678.1
56.5
13.4
10



AD-570679.1
98.0
15.0
10



AD-570680.1
62.4
17.6
10



AD-570681.1
44.9
2.0
10



AD-570682.1
23.9
7.9
10



AD-570717.1
112.1
3.8
10



AD-570963.1
54.1
14.8
10



AD-571157.1
70.6
5.8
10



AD-571158.1
60.8
9.7
10



AD-571168.1
112.1
27.3
10



AD-571298.1
18.4
3.4
10



AD-571298.2
16.1
0.7
10



AD-571447.1
24.4
1.2
10



AD-571448.1
30.6
0.3
10



AD-571449.1
106.0
22.3
10



AD-571539.4
27.8
4.7
10



AD-571719.1
22.6
1.5
10



AD-571752.3
27.6
7.7
10



AD-571753.1
16.1
1.6
10



AD-571765.1
64.1
15.8
10



AD-571766.1
55.0
5.8
10



AD-571767.1
32.1
5.8
10



AD-571825.1
17.6
3.5
10



AD-571826.1
19.8
3.3
10



AD-571900.1
44.3
5.2
10



AD-571945.1
29.3
3.3
10



AD-571948.1
58.6
14.3
10



AD-572039.3
60.4
0.9
10



AD-572040.3
27.6
8.2
10



AD-572041.3
34.5
1.6
10



AD-572044.1
46.2
3.5
10



AD-572049.1
45.8
4.6
10



AD-572060.1
55.6
6.5
10



AD-572061.2
33.3
3.7
10



AD-572062.3
27.7
0.3
10



AD-572108.1
116.7
22.8
10



AD-572235.1
13.6
3.8
10



AD-572258.1
21.1
6.0
10



AD-572278.1
26.8
10.1
10



AD-572279.1
23.4
5.7
10



AD-572281.1
16.1
3.0
10



AD-572355.1
126.5
3.3
10



AD-572356.1
15.0
3.9
10



AD-57238.2
18.4
3.6
10



AD-572387.1
26.8
1.2
10



AD-572388.4
32.0
8.3
10



AD-572389.3
23.9
3.5
10



AD-572390.2
27.7
1.0
10



AD-572393.1
33.8
3.3
10



AD-572613.1
59.1
12.2
10



AD-572614.1
45.4
12.3
10



AD-572858.1
34.6
0.7
10



AD-80806.9
18.8
2.1
10



AD-890084.1
10.0
2.2
10



AD-890085.1
73.2
12.5
10



AD-568976.1
43.6
6.6
1



AD-568978.1
37.1
4.4
1



AD-569127.1
57.8
6.9
1



AD-569133.1
28.9
2.8
1



AD-569164.3
36.1
8.2
1



AD-569164.4
66.7
7.7
1



AD-569195.1
47.7
2.3
1



AD-569237.1
104.2
16.9
1



AD-569239.1
97.9
0.6
1



AD-569272.3
83.3
2.5
1



AD-569350.1
96.0
8.6
1



AD-569571.1
45.3
1.7
1



AD-569763.3
30.4
10.2
1



AD-569764.1
60.6
10.0
1



AD-569766.1
97.0
10.7
1



AD-569816.1
35.8
2.7
1



AD-570156.1
50.7
7.8
1



AD-570466.1
47.2
10.5
1



AD-570470.1
104.5
9.3
1



AD-570471.1
79.8
8.0
1



AD-570474.1
71.1
13.3
1



AD-570475.1
96.1
5.0
1



AD-570476.1
47.1
4.0
1



AD-570620.1
33.8
5.0
1



AD-570621.1
50.0
5.5
1



AD-570622.1
23.0
1.3
1



AD-570623.1
25.8
2.6
1



AD-570624.1
24.5
4.1
1



AD-570625.1
42.3
7.6
1



AD-570627.1
46.6
1.6
1



AD-570631.1
71.3
6.1
1



AD-570632.1
51.7
4.1
1



AD-570672.1
55.4
3.5
1



AD-570674.1
49.1
7.1
1



AD-570675.1
79.1
5.3
1



AD-570676.1
104.9
3.3
1



AD-570677.1
81.2
3.2
1



AD-570678.1
88.9
15.3
1



AD-570679.1
47.1
8.1
1



AD-570680.1
65.2
2.9
1



AD-570681.1
68.2
4.0
1



AD-570682.1
59.1
8.0
1



AD-570717.1
67.5
7.9
1



AD-570963.1
83.7
1.0
1



AD-571157.1
103.6
15.4
1



AD-571158.1
83.5
11.5
1



AD-571168.1
95.5
5.4
1



AD-571298.1
29.0
9.5
1



AD-571298.2
26.7
2.1
1



AD-571447.1
83.8
7.0
1



AD-571448.1
72.5
5.6
1



AD-571449.1
85.6
8.0
1



AD-571539.4
47.7
4.8
1



AD-571719.1
23.6
4.3
1



AD-571752.3
69.3
9.5
1



AD-571753.1
37.9
6.5
1



AD-571765.1
65.3
3.2
1



AD-571766.1
56.3
9.7
1



AD-571767.1
30.4
9.6
1



AD-571825.1
19.5
4.7
1



AD-571826.1
24.2
3.6
1



AD-571900.1
55.9
4.3
1



AD-571945.1
31.2
1.6
1



AD-571948.1
91.5
19.5
1



AD-572039.3
86.5
8.4
1



AD-572040.3
65.8
2.2
1



AD-572041.3
41.5
4.4
1



AD-572044.1
60.9
0.8
1



AD-572049.1
60.4
0.9
1



AD-572060.1
68.9
6.1
1



AD-572061.2
42.7
4.7
1



AD-572062.3
27.5
6.5
1



AD-572108.1
82.1
10.1
1



AD-572235.1
21.6
2.5
1



AD-572258.1
30.4
5.4
1



AD-572278.1
22.1
3.9
1



AD-572279.1
37.0
6.4
1



AD-572281.1
26.6
1.5
1



AD-572355.1
88.8
17.7
1



AD-572356.1
57.4
16.2
1



AD-57238.2
47.0
7.0
1



AD-572387.1
37.9
2.5
1



AD-572388.4
25.7
3.3
1



AD-572389.3
28.1
4.4
1



AD-572390.2
36.4
1.6
1



AD-572393.1
52.1
2.7
1



AD-572613.1
95.0
6.4
1



AD-572614.1
60.8
1.1
1



AD-572858.1
46.6
0.3
1



AD-80806.9
27.0
3.4
1



AD-890084.1
23.3
6.5
1



AD-890085.1
109.4
8.4
1



AD-568976.1
61.6
17.4
0.1



AD-568978.1
81.5
7.4
0.1



AD-569127.1
93.9
18.4
0.1



AD-569133.1
55.0
7.4
0.1



AD-569164.3
77.5
20.5
0.1



AD-569164.4
93.7
3.2
0.1



AD-569195.1
89.6
2.7
0.1



AD-569237.1
110.5
13.2
0.1



AD-569239.1
108.4
2.2
0.1



AD-569272.3
89.2
13.7
0.1



AD-569350.1
96.1
10.9
0.1



AD-569571.1
91.2
11.2
0.1



AD-569763.3
87.3
9.1
0.1



AD-569764.1
88.7
7.7
0.1



AD-569766.1
103.3
10.3
0.1



AD-569816.1
81.0
8.2
0.1



AD-570156.1
81.4
9.9
0.1



AD-570466.1
87.4
1.5
0.1



AD-570470.1
100.2
12.6
0.1



AD-570471.1
96.4
4.0
0.1



AD-570474.1
95.0
6.4
0.1



AD-570475.1
104.7
2.8
0.1



AD-570476.1
88.1
13.9
0.1



AD-570620.1
56.3
8.1
0.1



AD-570621.1
93.7
24.7
0.1



AD-570622.1
61.7
13.5
0.1



AD-570623.1
75.4
4.9
0.1



AD-570624.1
80.8
6.3
0.1



AD-570625.1
90.4
6.4
0.1



AD-570627.1
89.3
6.8
0.1



AD-570631.1
91.6
8.4
0.1



AD-570632.1
86.5
7.7
0.1



AD-570672.1
78.1
12.7
0.1



AD-570674.1
90.8
7.5
0.1



AD-570675.1
94.8
6.1
0.1



AD-570676.1
101.1
0.7
0.1



AD-570677.1
88.5
15.2
0.1



AD-570678.1
95.4
4.1
0.1



AD-570679.1
100.5
8.2
0.1



AD-570680.1
100.0
3.6
0.1



AD-570681.1
70.3
14.5
0.1



AD-570682.1
94.8
9.0
0.1



AD-570717.1
98.8
8.1
0.1



AD-570963.1
97.1
8.0
0.1



AD-571157.1
94.0
10.5
0.1



AD-571158.1
98.6
7.3
0.1



AD-571168.1
103.7
8.9
0.1



AD-571298.1
56.5
9.3
0.1



AD-571298.2
46.2
12.6
0.1



AD-571447.1
111.3
8.3
0.1



AD-571448.1
98.9
6.9
0.1



AD-571449.1
101.0
4.6
0.1



AD-571539.4
86.3
9.2
0.1



AD-571719.1
69.1
5.8
0.1



AD-571752.3
93.8
25.2
0.1



AD-571753.1
86.2
12.6
0.1



AD-571765.1
100.3
9.3
0.1



AD-571766.1
92.0
16.7
0.1



AD-571767.1
87.6
3.3
0.1



AD-571825.1
36.2
7.2
0.1



AD-571826.1
64.0
8.1
0.1



AD-571900.1
94.0
8.3
0.1



AD-571945.1
85.9
5.5
0.1



AD-571948.1
91.7
8.5
0.1



AD-572039.3
118.3
9.9
0.1



AD-572040.3
90.6
9.6
0.1



AD-572041.3
81.0
7.3
0.1



AD-572044.1
94.0
0.3
0.1



AD-572049.1
100.1
11.7
0.1



AD-572060.1
94.7
6.8
0.1



AD-572061.2
78.4
3.2
0.1



AD-572062.3
91.7
14.2
0.1



AD-572108.1
93.7
10.4
0.1



AD-572235.1
70.4
10.5
0.1



AD-572258.1
68.0
3.6
0.1



AD-572278.1
80.0
9.0
0.1



AD-572279.1
78.6
4.9
0.1



AD-572281.1
66.7
3.6
0.1



AD-572355.1
101.9
7.5
0.1



AD-572356.1
85.5
8.2
0.1



AD-57238.2
81.2
13.9
0.1



AD-572387.1
90.3
1.0
0.1



AD-572388.4
76.1
12.0
0.1



AD-572389.3
81.1
13.6
0.1



AD-572390.2
88.8
1.2
0.1



AD-572393.1
86.4
1.5
0.1



AD-572613.1
101.3
16.5
0.1



AD-572614.1
95.5
3.1
0.1



AD-572858.1
78.1
19.1
0.1



AD-80806.9
61.6
1.7
0.1



AD-890084.1
73.7
9.0
0.1



AD-890085.1
109.0
13.9
0.1
















TABLE 13







C3 Single Dose Screens in PMH cells













Avg % C3






mRNA

Dose



Duplex
Remaining
SD
nM















AD-568976.1
3.0
0.7
50



AD-568978.1
2.1
0.2
50



AD-569127.1
15.8
1.0
50



AD-569133.1
70.4
29.5
50



AD-569164.3
69.0
20.9
50



AD-569164.4
75.4
22.3
50



AD-569195.1
81.9
25.8
50



AD-569237.1
207.6
49.5
50



AD-569239.1
161.6
51.4
50



AD-569272.3
101.8
23.8
50



AD-569350.1
146.4
53.4
50



AD-569571.1
23.8
6.6
50



AD-569763.3
57.4
28.9
50



AD-569764.1
22.3
6.5
50



AD-569766.1
159.6
28.7
50



AD-569816.1
59.9
19.5
50



AD-570156.1
26.6
11.5
50



AD-570466.1
81.9
3.3
50



AD-570470.1
140.3
51.7
50



AD-570471.1
121.3
43.9
50



AD-570474.1
139.2
57.9
50



AD-570475.1
119.7
54.3
50



AD-570476.1
77.5
1.6
50



AD-570620.1
13.3
0.1
50



AD-570621.1
52.4
16.5
50



AD-570622.1
13.9
1.8
50



AD-570623.1
15.3
1.2
50



AD-570624.1
50.7
5.1
50



AD-570625.1
27.6
2.1
50



AD-570627.1
36.8
1.7
50



AD-570631.1
103.0
5.0
50



AD-570632.1
89.5
19.1
50



AD-570672.1
66.0
13.2
50



AD-570674.1
118.1
35.4
50



AD-570675.1
210.6
49.7
50



AD-570676.1
151.5
34.6
50



AD-570677.1
116.2
32.0
50



AD-570678.1
194.9
9.9
50



AD-570679.1
128.4
56.7
50



AD-570680.1
135.9
47.7
50



AD-570681.1
84.0
7.4
50



AD-570682.1
107.7
37.1
50



AD-570717.1
165.7
61.6
50



AD-570963.1
113.2
32.5
50



AD-571157.1
140.6
8.0
50



AD-571158.1
179.6
62.3
50



AD-571168.1
144.1
56.1
50



AD-571298.1
2.0
0.2
50



AD-571298.2
1.0
0.2
50



AD-571447.1
133.2
53.5
50



AD-571448.1
109.2
34.9
50



AD-571449.1
164.6
61.6
50



AD-571539.4
73.3
1.1
50



AD-571719.1
5.2
1.4
50



AD-571752.3
115.0
23.3
50



AD-571753.1
23.1
3.4
50



AD-571765.1
121.3
19.5
50



AD-571766.1
94.8
30.2
50



AD-571767.1
88.0
32.8
50



AD-571825.1
7.7
1.7
50



AD-571826.1
18.2
5.0
50



AD-571900.1
92.7
28.1
50



AD-571945.1
85.7
10.8
50



AD-571948.1
169.3
87.5
50



AD-572039.3
118.5
58.9
50



AD-572040.3
105.6
4.1
50



AD-572041.3
101.1
1.8
50



AD-572044.1
97.4
11.5
50



AD-572049.1
116.5
18.6
50



AD-572060.1
99.7
3.2
50



AD-572061.2
90.6
2.4
50



AD-572062.3
83.1
31.2
50



AD-572108.1
141.0
12.3
50



AD-572235.1
21.0
1.1
50



AD-572258.1
4.5
1.3
50



AD-572278.1
2.6
0.1
50



AD-572279.1
2.5
0.6
50



AD-572281.1
2.0
0.5
50



AD-572355.1
159.0
48.4
50



AD-572356.1
78.3
12.7
50



AD-57238.2
9.9
0.8
50



AD-572387.1
9.0
1.9
50



AD-572388.4
4.3
1.2
50



AD-572389.3
1.8
0.7
50



AD-572390.2
5.8
2.3
50



AD-572393.1
31.9
6.1
50



AD-572613.1
217.6
101.2
50



AD-572614.1
116.3
22.1
50



AD-572858.1
107.6
42.0
50



AD-80806.9
1.1
0.2
50



AD-890084.1
13.1
5.5
50



AD-890085.1
121.6
20.3
50



AD-568976.1
10.5
1.8
10



AD-568978.1
9.8
5.4
10



AD-569127.1
52.8
11.1
10



AD-569133.1
116.3
31.4
10



AD-569164.3
99.7
7.5
10



AD-569164.4
42.7
3.8
10



AD-569195.1
117.9
47.1
10



AD-569237.1
177.3
6.2
10



AD-569239.1
154.2
30.6
10



AD-569272.3
122.8
24.2
10



AD-569350.1
71.4
11.6
10



AD-569571.1
20.8
5.4
10



AD-569763.3
31.1
9.9
10



AD-569764.1
62.8
26.7
10



AD-569766.1
158.6
21.9
10



AD-569816.1
61.8
22.2
10



AD-570156.1
35.0
6.6
10



AD-570466.1
149.7
29.3
10



AD-570470.1
138.8
45.5
10



AD-570471.1
59.6
5.4
10



AD-570474.1
61.0
0.4
10



AD-570475.1
68.6
12.7
10



AD-570476.1
93.3
11.6
10



AD-570620.1
50.2
13.3
10



AD-570621.1
102.6
12.3
10



AD-570622.1
78.7
22.3
10



AD-570623.1
45.0
13.8
10



AD-570624.1
115.2
43.3
10



AD-570625.1
85.5
10.7
10



AD-570627.1
111.1
16.7
10



AD-570631.1
69.7
22.4
10



AD-570632.1
96.7
21.6
10



AD-570672.1
68.9
14.1
10



AD-570674.1
150.8
33.1
10



AD-570675.1
170.0
28.6
10



AD-570676.1
152.1
4.7
10



AD-570677.1
203.3
10.3
10



AD-570678.1
190.5
30.9
10



AD-570679.1
209.3
45.6
10



AD-570680.1
169.1
17.7
10



AD-570681.1
116.0
26.5
10



AD-570682.1
118.6
33.8
10



AD-570717.1
198.1
4.5
10



AD-570963.1
97.4
31.4
10



AD-571157.1
72.7
8.0
10



AD-571158.1
57.4
4.9
10



AD-571168.1
57.9
6.1
10



AD-571298.1
5.7
1.7
10



AD-571298.2
2.7
1.0
10



AD-571447.1
187.9
30.2
10



AD-571448.1
55.4
7.1
10



AD-571449.1
174.5
53.4
10



AD-571539.4
124.8
50.3
10



AD-571719.1
22.7
5.7
10



AD-571752.3
54.4
5.9
10



AD-571753.1
91.4
12.2
10



AD-571765.1
92.9
33.3
10



AD-571766.1
57.0
3.6
10



AD-571767.1
50.5
5.8
10



AD-571825.1
27.0
7.4
10



AD-571826.1
18.1
3.0
10



AD-571900.1
71.4
11.9
10



AD-571945.1
96.8
7.3
10



AD-571948.1
119.7
27.4
10



AD-572039.3
117.5
18.2
10



AD-572040.3
169.3
47.8
10



AD-572041.3
134.4
44.7
10



AD-572044.1
159.2
22.4
10



AD-572049.1
57.7
6.2
10



AD-572060.1
170.5
7.1
10



AD-572061.2
144.3
31.5
10



AD-572062.3
96.8
37.4
10



AD-572108.1
54.9
5.8
10



AD-572235.1
77.9
44.2
10



AD-572258.1
18.0
4.6
10



AD-572278.1
10.7
3.2
10



AD-572279.1
11.3
5.8
10



AD-572281.1
7.2
0.6
10



AD-572355.1
57.0
6.3
10



AD-572356.1
56.4
6.0
10



AD-57238.2
39.9
3.8
10



AD-572387.1
25.3
10.0
10



AD-572388.4
25.3
7.5
10



AD-572389.3
4.0
0.6
10



AD-572390.2
25.0
4.1
10



AD-572393.1
102.7
20.6
10



AD-572613.1
150.3
34.6
10



AD-572614.1
139.9
44.5
10



AD-572858.1
54.4
5.3
10



AD-80806.9
1.4
0.5
10



AD-890084.1
42.3
7.2
10



AD-890085.1
151.9
21.0
10



AD-568976.1
56.6
32.8
1



AD-568978.1
46.8
16.6
1



AD-569127.1
46.2
2.3
1



AD-569133.1
109.6
22.6
1



AD-569164.3
99.8
16.0
1



AD-569164.4
39.9
1.3
1



AD-569195.1
73.1
28.2
1



AD-569237.1
86.5
26.9
1



AD-569239.1
115.6
17.9
1



AD-569272.3
117.3
13.3
1



AD-569350.1
123.4
21.0
1



AD-569571.1
77.2
28.3
1



AD-569763.3
96.4
22.9
1



AD-569764.1
107.4
7.7
1



AD-569766.1
72.0
37.4
1



AD-569816.1
84.3
29.9
1



AD-570156.1
77.2
11.3
1



AD-570466.1
112.4
31.4
1



AD-570470.1
87.9
18.5
1



AD-570471.1
95.2
8.7
1



AD-570474.1
100.2
21.4
1



AD-570475.1
100.1
17.1
1



AD-570476.1
65.5
4.2
1



AD-570620.1
88.9
22.3
1



AD-570621.1
114.1
57.6
1



AD-570622.1
118.7
26.7
1



AD-570623.1
107.4
25.7
1



AD-570624.1
100.8
23.8
1



AD-570625.1
134.9
17.5
1



AD-570627.1
117.1
19.9
1



AD-570631.1
67.0
1.7
1



AD-570632.1
78.9
17.5
1



AD-570672.1
85.0
25.5
1



AD-570674.1
92.1
28.0
1



AD-570675.1
127.1
18.9
1



AD-570676.1
111.7
28.9
1



AD-570677.1
139.7
35.4
1



AD-570678.1
150.4
15.1
1



AD-570679.1
76.8
12.4
1



AD-570680.1
98.3
14.7
1



AD-570681.1
110.4
10.0
1



AD-570682.1
66.0
15.0
1



AD-570717.1
99.7
8.4
1



AD-570963.1
132.6
25.3
1



AD-571157.1
116.5
18.5
1



AD-571158.1
117.7
23.5
1



AD-571168.1
97.9
10.8
1



AD-571298.1
22.6
12.7
1



AD-571298.2
13.0
3.1
1



AD-571447.1
100.3
4.7
1



AD-571448.1
83.5
12.5
1



AD-571449.1
64.9
9.1
1



AD-571539.4
94.1
20.2
1



AD-571719.1
81.1
35.0
1



AD-571752.3
93.9
17.5
1



AD-571753.1
59.7
12.0
1



AD-571765.1
114.3
18.7
1



AD-571766.1
105.2
10.6
1



AD-571767.1
111.3
22.5
1



AD-571825.1
95.5
6.9
1



AD-571826.1
94.3
20.3
1



AD-571900.1
105.4
22.4
1



AD-571945.1
104.8
17.4
1



AD-571948.1
104.1
21.3
1



AD-572039.3
135.4
11.0
1



AD-572040.3
128.9
26.4
1



AD-572041.3
115.9
43.0
1



AD-572044.1
112.3
6.8
1



AD-572049.1
86.1
12.8
1



AD-572060.1
133.9
13.8
1



AD-572061.2
137.5
3.0
1



AD-572062.3
86.9
5.7
1



AD-572108.1
109.8
25.8
1



AD-572235.1
75.6
17.8
1



AD-572258.1
36.8
7.4
1



AD-572278.1
49.8
16.2
1



AD-572279.1
73.8
28.3
1



AD-572281.1
56.8
13.9
1



AD-572355.1
96.9
13.9
1



AD-572356.1
95.9
11.2
1



AD-57238.2
132.4
20.9
1



AD-572387.1
60.5
21.8
1



AD-572388.4
39.8
10.3
1



AD-572389.3
26.0
7.1
1



AD-572390.2
88.5
25.7
1



AD-572393.1
114.8
25.2
1



AD-572613.1
82.7
16.4
1



AD-572614.1
121.5
9.4
1



AD-572858.1
90.8
9.7
1



AD-80806.9
6.1
2.3
1



AD-890084.1
90.9
24.6
1



AD-890085.1
108.3
63.0
1



AD-568976.1
108.7
10.5
0.1



AD-568978.1
89.4
17.2
0.1



AD-569127.1
113.6
35.6
0.1



AD-569133.1
83.3
16.5
0.1



AD-569164.3
103.9
28.8
0.1



AD-569164.4
112.7
28.0
0.1



AD-569195.1
148.7
14.3
0.1



AD-569237.1
123.3
25.7
0.1



AD-569239.1
108.0
13.5
0.1



AD-569272.3
107.5
14.8
0.1



AD-569350.1
117.1
27.8
0.1



AD-569571.1
107.2
30.7
0.1



AD-569763.3
163.9
11.1
0.1



AD-569764.1
73.1
10.8
0.1



AD-569766.1
152.3
13.3
0.1



AD-569816.1
118.5
24.7
0.1



AD-570156.1
124.5
32.6
0.1



AD-570466.1
103.6
25.5
0.1



AD-570470.1
140.4
34.3
0.1



AD-570471.1
124.0
35.8
0.1



AD-570474.1
103.0
24.7
0.1



AD-570475.1
90.4
10.1
0.1



AD-570476.1
132.6
22.6
0.1



AD-570620.1
129.3
46.6
0.1



AD-570621.1
116.8
5.5
0.1



AD-570622.1
109.1
17.6
0.1



AD-570623.1
130.5
15.8
0.1



AD-570624.1
92.6
14.7
0.1



AD-570625.1
103.8
3.9
0.1



AD-570627.1
99.9
0.5
0.1



AD-570631.1
120.9
21.2
0.1



AD-570632.1
124.5
21.6
0.1



AD-570672.1
116.3
15.7
0.1



AD-570674.1
80.7
13.7
0.1



AD-570675.1
106.4
38.0
0.1



AD-570676.1
83.4
16.8
0.1



AD-570677.1
138.1
5.4
0.1



AD-570678.1
103.1
16.3
0.1



AD-570679.1
81.6
11.9
0.1



AD-570680.1
121.7
20.3
0.1



AD-570681.1
111.4
18.4
0.1



AD-570682.1
128.5
22.4
0.1



AD-570717.1
129.3
36.1
0.1



AD-570963.1
129.7
28.9
0.1



AD-571157.1
115.1
2.3
0.1



AD-571158.1
131.7
29.6
0.1



AD-571168.1
132.0
42.0
0.1



AD-571298.1
81.0
15.3
0.1



AD-571298.2
116.1
18.1
0.1



AD-571447.1
142.9
60.2
0.1



AD-571448.1
94.5
28.3
0.1



AD-571449.1
137.8
18.9
0.1



AD-571539.4
126.8
43.6
0.1



AD-571719.1
95.0
22.0
0.1



AD-571752.3
127.5
28.5
0.1



AD-571753.1
142.2
39.1
0.1



AD-571765.1
127.6
31.8
0.1



AD-571766.1
161.2
16.9
0.1



AD-571767.1
191.4
8.6
0.1



AD-571825.1
132.2
37.6
0.1



AD-571826.1
156.2
52.6
0.1



AD-571900.1
135.3
24.6
0.1



AD-571945.1
99.6
8.3
0.1



AD-571948.1
80.1
14.9
0.1



AD-572039.3
138.5
13.3
0.1



AD-572040.3
140.2
7.2
0.1



AD-572041.3
110.9
27.0
0.1



AD-572044.1
111.8
14.5
0.1



AD-572049.1
160.6
39.0
0.1



AD-572060.1
113.3
18.8
0.1



AD-572061.2
114.8
21.0
0.1



AD-572062.3
131.5
32.8
0.1



AD-572108.1
150.8
23.6
0.1



AD-572235.1
80.3
11.2
0.1



AD-572258.1
88.5
1.9
0.1



AD-572278.1
99.5
19.6
0.1



AD-572279.1
99.8
32.6
0.1



AD-572281.1
108.0
7.9
0.1



AD-572355.1
130.0
19.3
0.1



AD-572356.1
131.8
29.0
0.1



AD-57238.2
89.6
32.9
0.1



AD-572387.1
136.2
34.6
0.1



AD-572388.4
100.6
10.7
0.1



AD-572389.3
98.0
21.8
0.1



AD-572390.2
123.9
37.7
0.1



AD-572393.1
132.4
45.2
0.1



AD-572613.1
126.0
25.0
0.1



AD-572614.1
78.8
11.9
0.1



AD-572858.1
103.7
19.5
0.1



AD-80806.9
27.8
3.0
0.1



AD-890084.1
152.2
33.6
0.1



AD-890085.1
112.9
8.8
0.1









Example 3. In Vivo Screening of dsRNA Duplexes in Mice

Duplexes of interest, identified from the above in vitro studies and shown in Table 15, were evaluated in vivo. In particular, at pre-dose day—14 wild-type mice (C57BL/6) were transduced by intravenous administration of 2×1011 viral particles of an adeno-associated virus 8 (AAV8) vector encoding human complement component C3. In particular, mice were administered an AAV8 encoding a portion of human complement component C3 mRNA spanning nucleotides 93-2893 of NM_000064.3, which includes a portion proximal to the 5′ UTR (referred to herein as AAV8.HsC3_p1), or an AAV8 encoding a portion of human complement component C3 mRNA spanning nucleotides 2293-4531 of NM_000064.3, which includes a portion of the 3′ UTR (referred to herein as AAV8.HsC3_p2).


At day 0, groups of three mice were subcutaneously administered a single 2 mg/kg dose of the agents of interest or PBS control. Table 14 provides the treatment groups and Table 15 provides the modifided nucleotide sequences of the sense and antisense strands of the duplexes of interest. At day 14 post-dose animals were sacrificed, liver samples were collected and snap-frozen in liquid nitrogen. Tissue mRNA was extracted and analyzed by the RT-QPCR method.


Human C3 mRNA levels were compared to housekeeping gene GAPDH. The values were then normalized to the average of PBS vehicle control group. The data were expressed as percent of baseline value, and presented as mean plus standard deviation. The results, listed in Table 16 and shown in FIG. 2, demonstrate that the exemplary duplex agents tested effectively reduce the level of the human C3 messenger RNA in vivo.













TABLE 14





Group
Animal





#
#
Treatment
AAV
Dose



















1
1
PBS
AAV8.HsC3_p1
2 mpk



2






3





2
4
Naïve





5






6





3
7
AD-569164.2





8






9





4
10
AD-569763.2





11






12





5
13
AD-565281.2





14






15





6
16
PBS
AAV8.HsC3_p2
2 mpk



17






18





7
19
Naïve





20






21





8
22
AD-571539.2





23






24





9
25
AD-572389.2





26






27





10
28
AD-567315.2





29






30





11
31
AD-571752.2





32






33





12
34
AD-568026.2





35






36





13
37
AD-572110.2





38






39





14
40
AD-572062.2





41






42





15
43
AD-572388.2





44






45





16
46
AD-572040.2





47






48





17
49
AD-567713.2





50






51





18
52
AD-567521.2





53






54





19
55
AD-567066.2





56






57




















TABLE 15








Nucleotide
SEQ





Sequence
ID


Duplex ID
Oligo ID
Strand
5′ to 3′
NO:







AD-569164.2
A-1085246.1
sense
asgsauccG
1070





faGfCfCfu






acuaugaau






L96




A-1093171.1
antis
asUfsucaU
1071





faGfUfagg






cUfcGfgau






cususc






AD-569763.2
A-1086444.1
sense
usgsggcaA
1072





fcUfCfCfa






acaauuacu






L96




A-1093754.1
antis
asGfsuaaU
1073





fuGfUfugg






aGfuUfgcc






cascsg






AD-565281.2
A-1085944.1
sense
csusaccaG
1074





faUfCfCfa






cuucaccau






L96




A-1085945.1
antis
asUfsggug
1075





(Agn)agug






gaUfcUfgg






uagsgsg






AD-571539.2
A-1089996.2
sense
ususccuuG
1076





faAfGfCfc






aacuacauu






L96




A-1095513.1
antis
asAfsuguA
1077





fgUfUfggc






uUfcAfagg






aasgsu






AD-572389.2
A-1091696.2
sense
asasggucU
1078





faCfGfCfc






uauuacaau






L96




A-1096354.1
antis
asUfsuguA
1079





faUfAfggc






gUfaGfacc






uusgsa






AD-567315.2
A-1090012.1
sense
asgsccaaC
1080





fuAfCfAfu






gaaccuacu






L96




A-1090013.1
antis
asGfsuagg
1081





(Tgn)ucau






guAfgUfug






gcususc






AD-571752.2
A-1090422.1
sense
uscsgugcG
1082





fuUfGfGfc






ucaaugaau






L96







A-1095726.1
antis
asUfsucaU
1083





fuGfAfgcc






aAfcGfcac






gascsg






AD-568026.2
A-1091434.1
sense
usgsgacaA
1084





faGfCfCfu






ucuccgauu






L96




A-1091435.1
antis
asAfsucgg
1085





(Agn)gaag






gcUfuUfgu






ccasgsc






AD-572110.2
A-1091138.1
sense
gsasugccA
1086





faGfAfAfc






acuaugauu






L96




A-1096084.1
antis
asAfsucaU
1087





faGfUfguu






cUfuGfgca






ucscsu






AD-572062.2
A-1091042.1
sense
csuscaagG
1088





fuCfAfCfc






auaaaaccu






L96







A-1096036.1
antis
asGfsguuU
1089





fuAfUfggu






gAfcCfuug






agsgsu






AD-572388.2
A-1091694.2
sense
csasagguC
1090





fuAfCfGfc






cuauuacau






L96




A-1096353.1
antis
asUfsguaA
1091





fuAfGfgcg






uAfgAfccu






ugsasc






AD-572040.2
A-1090998.2
sense
ascsucacC
1092





fuGfUfAfa






uaaauucgu






L96




A-1096014.1
antis
asCfsgaaU
1093





fuUfAfuua






cAfgGfuga






gususg






AD-567713.2
A-1090808.1
sense
ascscaagG
1094





faAfAfAfu






gaggguuuu






L96




A-1090809.1
antis
asAfsaacc
1095





(Cgn)ucau






uuUfcCfuu






gguscsu






AD-567521.2
A-1090424.1
sense
csgsugcgU
1096





fuGfGfCfu






caaugaacu






L96




A-1090425.1
antis
asGfsuuca
1097





(Tgn)ugag






ccAfaCfgc






acgsasc






AD-567066.2
A-1089514.1
sense
csgsugguC
1098





faAfGfGfu






cuucucucu






L96




A-1089515.1
antis
asGfsagag
1099





(Agn)agac






cuUfgAfcc






acgsusa



















TABLE 16






Duplex
Avg
SD



















PBS
100.10
5.09



Naïve
95.00
12.77



AD-569164.2
54.14
5.78



AD-569763.2
95.20
15.06



AD-565281.2
121.24
3.82



PBS
100.57
14.71



Naïve
87.32
20.75



AD-571539.2
89.52
11.77



AD-572389.2
73.16
14.10



AD-567315.2
90.15
22.27



AD-571752.2
87.97
28.36



AD-568026.2
150.52
13.23



AD-572110.2
86.55
10.98



AD-572062.2
104.01
0.90



AD-572388.2
71.83
23.25



AD-572040.2
107.74
50.53



AD-567713.2
149.76
7.94



AD-567521.2
85.10
23.93



AD-567066.2
101.62
0.28









Additional duplexes of interest, identified from the above in vitro studies and shown in Table 18, were also evaluated in vivo. In particular, at pre-dose day—14 wild-type mice (C57BL/6) were transduced by intravenous administration of 2×1011 viral particles of an adeno-associated virus 8 (AAV8) vector encoding human complement component C3.


At day 0, groups of three mice were subcutaneously administered a single 2 mg/kg dose of the agents of interest or PBS control. Table 17 provides the treatment groups and Table 18 provides the modifided nucleotide sequences of the sense and antisense strands of the duplexes of interest. At day 14 post-dose animals were sacrificed, liver samples were collected and snap-frozen in liquid nitrogen. Tissue mRNA was extracted and analyzed by the RT-QPCR method.


Human C3 mRNA levels were compared to housekeeping gene GAPDH. The values were then normalized to the average of PBS vehicle control group. The data were expressed as percent of baseline value, and presented as mean plus standard deviation. The results, listed in Table 19 and shown in FIG. 3, demonstrate that the exemplary duplex agents tested effectively reduce the level of the human C3 messenger RNA in vivo.













TABLE 17





Group
Animal





#
#
Treatment
AAV
Dose



















1
1
PBS
AAV8.HsC3_p1
2 mpk



2






3





2
4
Naïve





5






6





3
7
AD-565541.2





8






9





4
10
AD-569272.2





11






12





5
13
AD-569765.2





14






15





6
16
AD-564730.2





17






18





7
19
AD-564745.2





20






21





8
22
PBS
AAV8.HsC3_p2
2 mpk



23






24





9
25
Naïve





26






27





10
28
AD-571715.2





29






30





11
31
AD-572041.2





32






33





12
34
AD-572039.2





35






36





13
37
AD-568586.2





38






39





14
40
AD-566837.2





41






42





15
43
AD-566444.2





44






45





16
46
AD-567700.2





47






48





17
49
AD-567814.2





50






51





18
52
AD-568003.2





53






54




















TABLE 18








Nucleotide
SEQ 





Sequence
ID


Duplex ID
Oligo ID
Strand
5′ to 3′
NO:







AD-565541.2
A-1086464.1
sense
csasacaaU
1100





fuAfCfCfu






gcaucucuu






L96




A-1086465.1
antis
asAfsgaga
1101





(Tgn)gcag






guAfaUfug






uugsgsa






AD-569272.2
A-1085462.2
sense
asasuucuA
1102





fcUfAfCfa






ucuauaacu






L96




A-1093279.1
antis
asGfsuuaU
1103





faGfAfugu






aGfuAfgaa






uususc






AD-569765.2
A-1086448.1
sense
gsgscaacU
1104





fcCfAfAfc






aauuaccuu






L96




A-1093756.1
antis
asAfsgguA
1105





faUfUfguu






gGfaGfuug






ccscsa






AD-564730.2
A-1084842.1
sense
gsusaccuC
1106





fuUfCfAfu






ccagacagu






L96




A-1084843.1
antis
asCfsuguc
1107





(Tgn)ggau






gaAfgAfgg






uacscsc






AD-564745.2
A-1084872.1
sense
gsascagaC
1108





faAfGfAfc






caucuacau






L96




A-1084873.1
antis
asUfsguag
1109





(Agn)uggu






cuUfgUfcu






gucsusg






AD-571715.2
A-1090348.1
sense
csusacugC
1110





faGfCfUfa






aaagacuuu






L96




A-1095689.1
antis
asAfsaguC
1111





fuUfUfuag






cUfgCfagu






agsgsg






AD-572041.2
A-1091000.2
sense
csuscaccU
1112





fgUfAfAfu






aaauucgau






L96




A-1096015.1
antis
asUfscgaA
1113





fuUfUfauu






aCfaGfgug






agsusu






AD-572039.2
A-1090996.1
sense
asascucaC
1114





fcUfGfUfa






auaaauucu






L96




A-1096013.1
antis
asGfsaauU
1115





fuAfUfuac






aGfgUfgag






uusgsa






AD-568586.2
A-1092554.1
sense
gsasgaacC
1116





faGfAfAfa






caaugccau






L96




A-1092555.1
antis
asUfsggca
1117





(Tgn)uguu






ucUfgGfuu






cucsusu






AD-566837.2
A-1089056.1
sense
cscsgaguC
1118





fuGfAfGfa






ccagaauuu






L96




A-1089057.1
antis
asAfsauuc
1119





(Tgn)gguc






ucAfgAfcu






cggsusg






AD-566444.2
A-1088270.1
sense
ascsccuaC
1120





fuCfUfGfu






uguucgaau






L96




A-1088271.1
antis
asUfsucga
1121





(Agn)caac






agAfgUfag






ggusasg






AD-567700.2
A-1090782.1
sense
usgscgauC
1122





faGfAfAfg






agaccaagu






L96




A-1090783.1
antis
asCfsuugg
1123





(Tgn)cucu






ucUfgAfuc






gcasgsg






AD-567814.2
A-1091010.1
sense
csusguaaU
1124





faAfAfUfu






cgaccucau






L96




A-1091011.1
antis
asUfsgagg
1125





(Tgn)cgaa






uuUfaUfua






cagsgsu






AD-568003.2
A-1091388.1
sense
csasgauaC
1126





faUfCfUfc






caaguaugu






L96




A-1091389.1
antis
asCfsauac
1127





(Tgn)ugga






gaUfgUfau






cugsusc



















TABLE 19






Duplex
Avg
SD



















AD-565541.2
55.32
3.02



AD-569272.2
48.80
10.91



AD-569765.2
128.71
20.00



AD-564730.2
98.43
26.22



AD-564745.2
65.56
7.73



AD-571715.2
78.62
15.38



AD-572041.2
70.13
9.43



AD-572039.2
68.83
6.56



AD-568586.2
106.88
13.68



AD-566837.2
80.63
9.98



AD-566444.2
66.32
7.57



AD-567700.2
58.92
1.17



AD-567814.2
132.61
17.19



AD-568003.2
112.42
1.84









Example 4. Additional Duplexes Targeting Human C3

Additional agents targeting the human complement component C3 (C3) gene (human: NCBI refseqID NM_000064.3; NCBI GeneID: 718) were designed using custom R and Python scripts and synthesized as described above.


Detailed lists of the unmodified complement component C3 sense and antisense strand nucleotide sequences are shown in Tables 20 and 22. Detailed lists of the modified complement component C3 sense and antisense strand nucleotide sequences are shown in Tables 21 and 23.


Single dose screens of the additional agents were performed by free uptake and transfection.


For free uptake, experiments were performed by adding 2.5 μl of siRNA duplexes in PBS per well into a 96 well plate. Complete growth media (47.5 μl) containing about 1.5×104 primary cynomolgus hepatocytes (PCH) were then added to the siRNA. Cells were incubated for 48 hours prior to RNA purification and RT-qPCR. Single dose experiments were performed at 500 nM, 100 nM, and 10 nM final duplex concentration.


For transfections, 7.5 μl of Opti-MEM plus 0.1 μl of Lipofectamine RNAiMax per well (Invitrogen, Carlsbad CA. cat #13778-150) was added to 2.5 μl of each siRNA duplex to an individual well in a 384-well plate. The mixture was then incubated at room temperature for 15 minutes. Forty μl of complete growth media without antibiotic containing ˜1.5×104 primary cynomolgus hepatocytes (PCH) were then added to the siRNA mixture. Cells were incubated for 24 hours prior to RNA purification. Single dose experiments were performed at 50, nM, 10 nM, 1 nM, and 0.1 nM final duplex concentration.


Total RNA isolation was performed using DYNABEADS. Briefly, cells are lysed in 10 μl of Lysis/Binding Buffer containing 3 μL of beads per well are mixed for 10 minutes on an electrostatic shaker. The washing steps are automated on a Biotek EL406, using a magnetic plate support. Beads are washed (in 3 L) once in Buffer A, once in Buffer B, and twice in Buffer E, with aspiration steps in between. Following a final aspiration, complete 12 μL RT mixture is added to each well, as described below.


For cDNA synthesis, a master mix of 1.5 μl 10× Buffer, 0.6 μl 10× dNTPs, 1.5 μl Random primers, 0.75 μl Reverse Transcriptase, 0.75 μl RNase inhibitor and 9.9 μl of H2O per reaction were added per well. Plates were sealed, agitated for 10 minutes on an electrostatic shaker, and then incubated at 37 degrees C. for 2 hours. Following this, the plates were agitated at 80 degrees C. for 8 minutes.


RT-qPCR was performed as described above and relative fold change was calculated as described above.


The results of the free uptake experiments (FU) and the transfection experiments (TX) of the dsRNA agents in Tables 20 and 21 in PCH are shown in Tables 24-26. The results of the free uptake experiments (FU) and the transfection experiments (TX) of the dsRNA agents in Tables 22 and 23 in PCH are shown in Tables 27-29.









TABLE 20







Unmodified Sense and Antisense Strand Sequences of Complement Component C3 dsRNA Agents















SEQ
Range in

SEQ
Range in



Sense
ID
NM_000
Antisense
ID
NM_000


Duplex Name
Sequence 5′ to 3′
NO:
064.3
Sequence 5′ to 3′
NO:
064.3





AD-570137.1
GUGCUGAAUAAGAAGAACAAU
1128
1903-1923
AUUGUUCUUCUUAUUCAGCACGA
1393
1901-1923





AD-570138.1
UGCUGAAUAAGAAGAACAAAU
1129
1904-1924
AUUUGUUCUUCUUAUUCAGCACG
1394
1902-1924





AD-570139.1
GCUGAAUAAGAAGAACAAACU
1130
1905-1925
AGUUUGUUCUUCUUAUUCAGCAC
1395
1903-1925





AD-570140.1
CUGAAUAAGAAGAACAAACUU
1131
1906-1926
AAGUUUGUUCUUCUUAUUCAGCA
1396
1904-1926





AD-570141.1
UGAAUAAGAAGAACAAACUGU
1132
1907-1927
ACAGUUUGUUCUUCUUAUUCAGC
1397
1905-1927





AD-570142.1
GAAUAAGAAGAACAAACUGAU
1133
1908-1928
AUCAGUUUGUUCUUCUUAUUCAG
1398
1906-1928





AD-570143.1
AAUAAGAAGAACAAACUGACU
1134
1909-1929
AGUCAGUUUGUUCUUCUUAUUCA
1399
1907-1929





AD-570144.1
AUAAGAAGAACAAACUGACGU
1135
1910-1930
ACGUCAGUUUGUUCUUCUUAUUC
1400
1908-1930





AD-570145.1
UAAGAAGAACAAACUGACGCU
1136
1911-1931
AGCGUCAGUUUGUUCUUCUUAUU
1401
1909-1931





AD-570146.1
AAGAAGAACAAACUGACGCAU
1137
1912-1932
AUGCGUCAGUUUGUUCUUCUUAU
1402
1910-1932





AD-570147.1
AGAAGAACAAACUGACGCAGU
1138
1913-1933
ACUGCGUCAGUUUGUUCUUCUUA
1403
1911-1933





AD-570148.1
GAAGAACAAACUGACGCAGAU
1139
1914-1934
AUCUGCGUCAGUUUGUUCUUCUU
1404
1912-1934





AD-570149.1
AAGAACAAACUGACGCAGAGU
1140
1915-1935
ACUCUGCGUCAGUUUGUUCUUCU
1405
1913-1935





AD-570150.1
AGAACAAACUGACGCAGAGUU
1141
1916-1936
AACUCUGCGUCAGUUUGUUCUUC
1406
1914-1936





AD-570151.1
GAACAAACUGACGCAGAGUAU
1142
1917-1937
AUACUCUGCGUCAGUUUGUUCUU
1407
1915-1937





AD-570152.1
AACAAACUGACGCAGAGUAAU
1143
1918-1938
AUUACUCUGCGUCAGUUUGUUCU
1408
1916-1938





AD-570153.1
ACAAACUGACGCAGAGUAAGU
1144
1919-1939
ACUUACUCUGCGUCAGUUUGUUC
1409
1917-1939





AD-570154.1
CAAACUGACGCAGAGUAAGAU
1145
1920-1940
AUCUUACUCUGCGUCAGUUUGUU
1410
1918-1940





AD-570155.1
AAACUGACGCAGAGUAAGAUU
1146
1921-1941
AAUCUUACUCUGCGUCAGUUUGU
1411
1919-1941





AD-570156.2
AACUGACGCAGAGUAAGAUCU
1147
1922-1942
AGAUCUUACUCUGCGUCAGUUUG
1412
1920-1942





AD-570158.1
CUGACGCAGAGUAAGAUCUGU
1148
1924-1944
ACAGAUCUUACUCUGCGUCAGUU
1413
1922-1944





AD-570159.1
UGACGCAGAGUAAGAUCUGGU
1149
1925-1945
ACCAGAUCUUACUCUGCGUCAGU
1414
1923-1945





AD-570160.1
GACGCAGAGUAAGAUCUGGGU
1150
1926-1946
ACCCAGAUCUUACUCUGCGUCAG
1415
1924-1946





AD-570161.1
ACGCAGAGUAAGAUCUGGGAU
1151
1927-1947
AUCCCAGAUCUUACUCUGCGUCA
1416
1925-1947





AD-570611.1
UGAGCAUGUCGGACAAGAAAU
1152
2513-2533
AUUUCUUGUCCGACAUGCUCACA
1417
2511-2533





AD-570612.1
GAGCAUGUCGGACAAGAAAGU
1153
2514-2534
ACUUUCUUGUCCGACAUGCUCAC
1418
2512-2534





AD-570613.1
AGCAUGUCGGACAAGAAAGGU
1154
2515-2535
ACCUUUCUUGUCCGACAUGCUCA
1419
2513-2535





AD-570614.1
GCAUGUCGGACAAGAAAGGGU
1155
2516-2536
ACCCUUUCUUGUCCGACAUGCUC
1420
2514-2536





AD-570615.1
CAUGUCGGACAAGAAAGGGAU
1156
2517-2537
AUCCCUUUCUUGUCCGACAUGCU
1421
2515-2537





AD-570616.1
AUGUCGGACAAGAAAGGGAUU
1157
2518-2538
AAUCCCUUUCUUGUCCGACAUGC
1422
2516-2538





AD-570617.1
UGUCGGACAAGAAAGGGAUCU
1158
2519-2539
AGAUCCCUUUCUUGUCCGACAUG
1423
2517-2539





AD-570618.1
GUCGGACAAGAAAGGGAUCUU
1159
2520-2540
AAGAUCCCUUUCUUGUCCGACAU
1424
2518-2540





AD-570619.1
UCGGACAAGAAAGGGAUCUGU
1160
2521-2541
ACAGAUCCCUUUCUUGUCCGACA
1425
2519-2541





AD-570620.3
CGGACAAGAAAGGGAUCUGUU
1161
2522-2542
AACAGAUCCCUUUCUUGUCCGAC
1426
2520-2542





AD-570621.2
GGACAAGAAAGGGAUCUGUGU
1162
2523-2543
ACACAGAUCCCUUUCUUGUCCGA
1427
2521-2543





AD-570622.2
GACAAGAAAGGGAUCUGUGUU
1163
2524-2544
AACACAGAUCCCUUUCUUGUCCG
1428
2522-2544





AD-570623.4
ACAAGAAAGGGAUCUGUGUGU
1164
2525-2545
ACACACAGAUCCCUUUCUUGUCC
1429
2523-2545





AD-570624.2
CAAGAAAGGGAUCUGUGUGGU
1165
2526-2546
ACCACACAGAUCCCUUUCUUGUC
1430
2524-2546





AD-570625.2
AAGAAAGGGAUCUGUGUGGCU
1166
2527-2547
AGCCACACAGAUCCCUUUCUUGU
1431
2525-2547





AD-570626.1
AGAAAGGGAUCUGUGUGGCAU
1167
2528-2548
AUGCCACACAGAUCCCUUUCUUG
1432
2526-2548





AD-570627.2
GAAAGGGAUCUGUGUGGCAGU
1168
2529-2549
ACUGCCACACAGAUCCCUUUCUU
1433
2527-2549





AD-570628.1
AAAGGGAUCUGUGUGGCAGAU
1169
2530-2550
AUCUGCCACACAGAUCCCUUUCU
1434
2528-2550





AD-570629.1
AAGGGAUCUGUGUGGCAGACU
1170
2531-2551
AGUCUGCCACACAGAUCCCUUUC
1435
2529-2551





AD-570630.1
AGGGAUCUGUGUGGCAGACCU
1171
2532-2552
AGGUCUGCCACACAGAUCCCUUU
1436
2530-2552





AD-1069837.1
GGGAUCUGUGUGGCAGACCCU
1172
2500-2520
AGGGUCUGCCACACAGAUCCCUU
1437
2498-2520





AD-570707.1
GAAAUCCGAGCCGUUCUCUAU
1173
2629-2649
AUAGAGAACGGCUCGGAUUUCCA
1438
2627-2649





AD-570708.1
AAAUCCGAGCCGUUCUCUACU
1174
2630-2650
AGUAGAGAACGGCUCGGAUUUCC
1439
2628-2650





AD-570709.1
AAUCCGAGCCGUUCUCUACAU
1175
2631-2651
AUGUAGAGAACGGCUCGGAUUUC
1440
2629-2651





AD-570710.1
AUCCGAGCCGUUCUCUACAAU
1176
2632-2652
AUUGUAGAGAACGGCUCGGAUUU
1441
2630-2652





AD-570715.1
AGCCGUUCUCUACAAUUACCU
1177
2637-2657
AGGUAAUUGUAGAGAACGGCUCG
1442
2635-2657





AD-570716.1
GCCGUUCUCUACAAUUACCGU
1178
2638-2658
ACGGUAAUUGUAGAGAACGGCUC
1443
2636-2658





AD-570717.2
CCGUUCUCUACAAUUACCGGU
1179
2639-2659
ACCGGUAAUUGUAGAGAACGGCU
1444
2637-2659





AD-570718.1
CGUUCUCUACAAUUACCGGCU
1180
2640-2660
AGCCGGUAAUUGUAGAGAACGGC
1445
2638-2660





AD-570719.1
GUUCUCUACAAUUACCGGCAU
1181
2641-2661
AUGCCGGUAAUUGUAGAGAACGG
1446
2639-2661





AD-570720.1
UUCUCUACAAUUACCGGCAGU
1182
2642-2662
ACUGCCGGUAAUUGUAGAGAACG
1447
2640-2662





AD-570721.1
UCUCUACAAUUACCGGCAGAU
1183
2643-2663
AUCUGCCGGUAAUUGUAGAGAAC
1448
2641-2663





AD-571285.1
GGCUGACCGCCUACGUGGUCU
1184
3323-3343
AGACCACGUAGGCGGUCAGCCAG
1449
3321-3343





AD-571286.1
GCUGACCGCCUACGUGGUCAU
1185
3324-3344
AUGACCACGUAGGCGGUCAGCCA
1450
3322-3344





AD-571287.1
CUGACCGCCUACGUGGUCAAU
1186
3325-3345
AUUGACCACGUAGGCGGUCAGCC
1451
3323-3345





AD-571288.1
UGACCGCCUACGUGGUCAAGU
1187
3326-3346
ACUUGACCACGUAGGCGGUCAGC
1452
3324-3346





AD-571289.1
GACCGCCUACGUGGUCAAGGU
1188
3327-3347
ACCUUGACCACGUAGGCGGUCAG
1453
3325-3347





AD-571290.1
ACCGCCUACGUGGUCAAGGUU
1189
3328-3348
AACCUUGACCACGUAGGCGGUCA
1454
3326-3348





AD-571291.1
CCGCCUACGUGGUCAAGGUCU
1190
3329-3349
AGACCUUGACCACGUAGGCGGUC
1455
3327-3349





AD-571292.1
CGCCUACGUGGUCAAGGUCUU
1191
3330-3350
AAGACCUUGACCACGUAGGCGGU
1456
3328-3350





AD-571293.1
GCCUACGUGGUCAAGGUCUUU
1192
3331-3351
AAAGACCUUGACCACGUAGGCGG
1457
3329-3351





AD-571294.1
CCUACGUGGUCAAGGUCUUCU
1193
3332-3352
AGAAGACCUUGACCACGUAGGCG
1458
3330-3352





AD-571295.1
CUACGUGGUCAAGGUCUUCUU
1194
3333-3353
AAGAAGACCUUGACCACGUAGGC
1459
3331-3353





AD-571296.1
UACGUGGUCAAGGUCUUCUCU
1195
3334-3354
AGAGAAGACCUUGACCACGUAGG
1460
3332-3354





AD-571297.1
ACGUGGUCAAGGUCUUCUCUU
1196
3335-3355
AAGAGAAGACCUUGACCACGUAG
1461
3333-3355





AD-571298.6
CGUGGUCAAGGUCUUCUCUCU
1197
3336-3356
AGAGAGAAGACCUUGACCACGUA
1462
3334-3356





AD-571299.1
GUGGUCAAGGUCUUCUCUCUU
1198
3337-3357
AAGAGAGAAGACCUUGACCACGU
1463
3335-3357





AD-571300.1
UGGUCAAGGUCUUCUCUCUGU
1199
3338-3358
ACAGAGAGAAGACCUUGACCACG
1464
3336-3358





AD-571301.1
GGUCAAGGUCUUCUCUCUGGU
1200
3339-3359
ACCAGAGAGAAGACCUUGACCAC
1465
3337-3359





AD-571302.1
GUCAAGGUCUUCUCUCUGGCU
1201
3340-3360
AGCCAGAGAGAAGACCUUGACCA
1466
3338-3360





AD-571303.1
UCAAGGUCUUCUCUCUGGCUU
1202
3341-3361
AAGCCAGAGAGAAGACCUUGACC
1467
3339-3361





AD-571304.1
CAAGGUCUUCUCUCUGGCUGU
1203
3342-3362
ACAGCCAGAGAGAAGACCUUGAC
1468
3340-3362





AD-571305.1
AAGGUCUUCUCUCUGGCUGUU
1204
3343-3363
AACAGCCAGAGAGAAGACCUUGA
1469
3341-3363





AD-571306.1
AGGUCUUCUCUCUGGCUGUCU
1205
3344-3364
AGACAGCCAGAGAGAAGACCUUG
1470
3342-3364





AD-571307.1
GGUCUUCUCUCUGGCUGUCAU
1206
3345-3365
AUGACAGCCAGAGAGAAGACCUU
1471
3343-3365





AD-571308.1
GUCUUCUCUCUGGCUGUCAAU
1207
3346-3366
AUUGACAGCCAGAGAGAAGACCU
1472
3344-3366





AD-571309.1
UCUUCUCUCUGGCUGUCAACU
1208
3347-3367
AGUUGACAGCCAGAGAGAAGACC
1473
3345-3367





AD-571526.1
UAAAGCAGGAGACUUCCUUGU
1209
3603-3623
ACAAGGAAGUCUCCUGCUUUAGU
1474
3601-3623





AD-571527.1
AAAGCAGGAGACUUCCUUGAU
1210
3604-3624
AUCAAGGAAGUCUCCUGCUUUAG
1475
3602-3624





AD-571528.1
AAGCAGGAGACUUCCUUGAAU
1211
3605-3625
AUUCAAGGAAGUCUCCUGCUUUA
1476
3603-3625





AD-571529.1
AGCAGGAGACUUCCUUGAAGU
1212
3606-3626
ACUUCAAGGAAGUCUCCUGCUUU
1477
3604-3626





AD-571530.1
GCAGGAGACUUCCUUGAAGCU
1213
3607-3627
AGCUUCAAGGAAGUCUCCUGCUU
1478
3605-3627





AD-571531.1
CAGGAGACUUCCUUGAAGCCU
1214
3608-3628
AGGCUUCAAGGAAGUCUCCUGCU
1479
3606-3628





AD-571532.1
AGGAGACUUCCUUGAAGCCAU
1215
3609-3629
AUGGCUUCAAGGAAGUCUCCUGC
1480
3607-3629





AD-571533.1
GGAGACUUCCUUGAAGCCAAU
1216
3610-3630
AUUGGCUUCAAGGAAGUCUCCUG
1481
3608-3630





AD-571534.1
GAGACUUCCUUGAAGCCAACU
1217
3611-3631
AGUUGGCUUCAAGGAAGUCUCCU
1482
3609-3631





AD-568955.1
AGAGCGGGUACCUCUUCAUCU
1218
470-490
AGAUGAAGAGGUACCCGCUCUGC
1483
468-490





AD-568956.1
GAGCGGGUACCUCUUCAUCCU
1219
471-491
AGGAUGAAGAGGUACCCGCUCUG
1484
469-491





AD-568957.1
AGCGGGUACCUCUUCAUCCAU
1220
472-492
AUGGAUGAAGAGGUACCCGCUCU
1485
470-492





AD-568958.1
GCGGGUACCUCUUCAUCCAGU
1221
473-493
ACUGGAUGAAGAGGUACCCGCUC
1486
471-493





AD-568959.1
CGGGUACCUCUUCAUCCAGAU
1222
474-494
AUCUGGAUGAAGAGGUACCCGCU
1487
472-494





AD-568960.1
GGGUACCUCUUCAUCCAGACU
1223
475-495
AGUCUGGAUGAAGAGGUACCCGC
1488
473-495





AD-568961.1
GGUACCUCUUCAUCCAGACAU
1224
476-496
AUGUCUGGAUGAAGAGGUACCCG
1489
474-496





AD-568962.1
GUACCUCUUCAUCCAGACAGU
1225
477-497
ACUGUCUGGAUGAAGAGGUACCC
1490
475-497





AD-568963.2
UACCUCUUCAUCCAGACAGAU
1226
478-498
AUCUGUCUGGAUGAAGAGGUACC
1491
476-498





AD-568964.1
ACCUCUUCAUCCAGACAGACU
1227
479-499
AGUCUGUCUGGAUGAAGAGGUAC
1492
477-499





AD-568965.1
CCUCUUCAUCCAGACAGACAU
1228
480-500
AUGUCUGUCUGGAUGAAGAGGUA
1493
478-500





AD-568966.1
CUCUUCAUCCAGACAGACAAU
1229
481-501
AUUGUCUGUCUGGAUGAAGAGGU
1494
479-501





AD-568967.1
UCUUCAUCCAGACAGACAAGU
1230
482-502
ACUUGUCUGUCUGGAUGAAGAGG
1495
480-502





AD-568968.1
CUUCAUCCAGACAGACAAGAU
1231
483-503
AUCUUGUCUGUCUGGAUGAAGAG
1496
481-503





AD-568969.1
UUCAUCCAGACAGACAAGACU
1232
484-504
AGUCUUGUCUGUCUGGAUGAAGA
1497
482-504





AD-568970.1
UCAUCCAGACAGACAAGACCU
1233
485-505
AGGUCUUGUCUGUCUGGAUGAAG
1498
483-505





AD-568971.1
CAUCCAGACAGACAAGACCAU
1234
486-506
AUGGUCUUGUCUGUCUGGAUGAA
1499
484-506





AD-568972.1
AUCCAGACAGACAAGACCAUU
1235
487-507
AAUGGUCUUGUCUGUCUGGAUGA
1500
485-507





AD-568973.1
UCCAGACAGACAAGACCAUCU
1236
488-508
AGAUGGUCUUGUCUGUCUGGAUG
1501
486-508





AD-568974.1
CCAGACAGACAAGACCAUCUU
1237
489-509
AAGAUGGUCUUGUCUGUCUGGAU
1502
487-509





AD-568975.1
CAGACAGACAAGACCAUCUAU
1238
490-510
AUAGAUGGUCUUGUCUGUCUGGA
1503
488-510





AD-568977.1
GACAGACAAGACCAUCUACAU
1239
492-512
AUGUAGAUGGUCUUGUCUGUCUG
1504
490-512





AD-568979.1
CAGACAAGACCAUCUACACCU
1240
494-514
AGGUGUAGAUGGUCUUGUCUGUC
1505
492-514





AD-1069834.1
AGACAAGACCAUCUACACCCU
1241
495-515
AGGGUGUAGAUGGUCUUGUCUGU
1506
493-515





AD-1069835.1
GACAAGACCAUCUACACCCCU
1242
496-516
AGGGGUGUAGAUGGUCUUGUCUG
1507
494-516





AD-1069836.1
ACAAGACCAUCUACACCCCUU
1243
497-517
AAGGGGUGUAGAUGGUCUUGUCU
1508
495-517





AD-569154.1
GGCCAGUGGAAGAUCCGAGCU
1244
697-717
AGCUCGGAUCUUCCACUGGCCCA
1509
695-717





AD-569155.1
GCCAGUGGAAGAUCCGAGCCU
1245
698-718
AGGCUCGGAUCUUCCACUGGCCC
1510
696-718





AD-569156.1
CCAGUGGAAGAUCCGAGCCUU
1246
699-719
AAGGCUCGGAUCUUCCACUGGCC
1511
697-719





AD-569157.1
CAGUGGAAGAUCCGAGCCUAU
1247
700-720
AUAGGCUCGGAUCUUCCACUGGC
1512
698-720





AD-569158.1
AGUGGAAGAUCCGAGCCUACU
1248
701-721
AGUAGGCUCGGAUCUUCCACUGG
1513
699-721





AD-569159.1
GUGGAAGAUCCGAGCCUACUU
1249
702-722
AAGUAGGCUCGGAUCUUCCACUG
1514
700-722





AD-569160.1
UGGAAGAUCCGAGCCUACUAU
1250
703-723
AUAGUAGGCUCGGAUCUUCCACU
1515
701-723





AD-569161.1
GGAAGAUCCGAGCCUACUAUU
1251
704-724
AAUAGUAGGCUCGGAUCUUCCAC
1516
702-724





AD-569162.1
GAAGAUCCGAGCCUACUAUGU
1252
705-725
ACAUAGUAGGCUCGGAUCUUCCA
1517
703-725





AD-569163.1
AAGAUCCGAGCCUACUAUGAU
1253
706-726
AUCAUAGUAGGCUCGGAUCUUCC
1518
704-726





AD-569166.1
AUCCGAGCCUACUAUGAAAAU
1254
709-729
AUUUUCAUAGUAGGCUCGGAUCU
1519
707-729





AD-569167.1
UCCGAGCCUACUAUGAAAACU
1255
710-730
AGUUUUCAUAGUAGGCUCGGAUC
1520
708-730





AD-569168.1
CCGAGCCUACUAUGAAAACUU
1256
711-731
AAGUUUUCAUAGUAGGCUCGGAU
1521
709-731





AD-569169.1
CGAGCCUACUAUGAAAACUCU
1257
712-732
AGAGUUUUCAUAGUAGGCUCGGA
1522
710-732





AD-569170.1
GAGCCUACUAUGAAAACUCAU
1258
713-733
AUGAGUUUUCAUAGUAGGCUCGG
1523
711-733





AD-569171.1
AGCCUACUAUGAAAACUCACU
1259
714-734
AGUGAGUUUUCAUAGUAGGCUCG
1524
712-734





AD-569172.1
GCCUACUAUGAAAACUCACCU
1260
715-735
AGGUGAGUUUUCAUAGUAGGCUC
1525
713-735





AD-569173.1
CCUACUAUGAAAACUCACCAU
1261
716-736
AUGGUGAGUUUUCAUAGUAGGCU
1526
714-736





AD-569174.1
CUACUAUGAAAACUCACCACU
1262
717-737
AGUGGUGAGUUUUCAUAGUAGGC
1527
715-737





AD-569175.1
UACUAUGAAAACUCACCACAU
1263
718-738
AUGUGGUGAGUUUUCAUAGUAGG
1528
716-738





AD-569262.1
CCUACAGAGAAAUUCUACUAU
1264
805-825
AUAGUAGAAUUUCUCUGUAGGCU
1529
803-825





AD-569263.1
CUACAGAGAAAUUCUACUACU
1265
806-826
AGUAGUAGAAUUUCUCUGUAGGC
1530
804-826





AD-569264.1
UACAGAGAAAUUCUACUACAU
1266
807-827
AUGUAGUAGAAUUUCUCUGUAGG
1531
805-827





AD-569265.1
ACAGAGAAAUUCUACUACAUU
1267
808-828
AAUGUAGUAGAAUUUCUCUGUAG
1532
806-828





AD-569266.1
CAGAGAAAUUCUACUACAUCU
1268
809-829
AGAUGUAGUAGAAUUUCUCUGUA
1533
807-829





AD-569267.1
AGAGAAAUUCUACUACAUCUU
1269
810-830
AAGAUGUAGUAGAAUUUCUCUGU
1534
808-830





AD-569268.1
GAGAAAUUCUACUACAUCUAU
1270
811-831
AUAGAUGUAGUAGAAUUUCUCUG
1535
809-831





AD-569269.1
AGAAAUUCUACUACAUCUAUU
1271
812-832
AAUAGAUGUAGUAGAAUUUCUCU
1536
810-832





AD-569270.1
GAAAUUCUACUACAUCUAUAU
1272
813-833
AUAUAGAUGUAGUAGAAUUUCUC
1537
811-833





AD-569271.1
AAAUUCUACUACAUCUAUAAU
1273
814-834
AUUAUAGAUGUAGUAGAAUUUCU
1538
812-834





AD-569273.1
AUUCUACUACAUCUAUAACGU
1274
816-836
ACGUUAUAGAUGUAGUAGAAUUU
1539
814-836





AD-569274.1
UUCUACUACAUCUAUAACGAU
1275
817-837
AUCGUUAUAGAUGUAGUAGAAUU
1540
815-837





AD-569275.1
UCUACUACAUCUAUAACGAGU
1276
818-838
ACUCGUUAUAGAUGUAGUAGAAU
1541
816-838





AD-569276.1
CUACUACAUCUAUAACGAGAU
1277
819-839
AUCUCGUUAUAGAUGUAGUAGAA
1542
817-839





AD-569277.1
UACUACAUCUAUAACGAGAAU
1278
820-840
AUUCUCGUUAUAGAUGUAGUAGA
1543
818-840





AD-569278.1
ACUACAUCUAUAACGAGAAGU
1279
821-841
ACUUCUCGUUAUAGAUGUAGUAG
1544
819-841





AD-569279.1
CUACAUCUAUAACGAGAAGGU
1280
822-842
ACCUUCUCGUUAUAGAUGUAGUA
1545
820-842





AD-569280.1
UACAUCUAUAACGAGAAGGGU
1281
823-843
ACCCUUCUCGUUAUAGAUGUAGU
1546
821-843





AD-569281.1
ACAUCUAUAACGAGAAGGGCU
1282
824-844
AGCCCUUCUCGUUAUAGAUGUAG
1547
822-844





AD-569282.1
CAUCUAUAACGAGAAGGGCCU
1283
825-845
AGGCCCUUCUCGUUAUAGAUGUA
1548
823-845





AD-569506.1
CCUCUCCCUACCAGAUCCACU
1284
1142-1162
AGUGGAUCUGGUAGGGAGAGGUC
1549
1140-1162





AD-569507.1
CUCUCCCUACCAGAUCCACUU
1285
1143-1163
AAGUGGAUCUGGUAGGGAGAGGU
1550
1141-1163





AD-569508.1
UCUCCCUACCAGAUCCACUUU
1286
1144-1164
AAAGUGGAUCUGGUAGGGAGAGG
1551
1142-1164





AD-569509.1
CUCCCUACCAGAUCCACUUCU
1287
1145-1165
AGAAGUGGAUCUGGUAGGGAGAG
1552
1143-1165





AD-569510.1
UCCCUACCAGAUCCACUUCAU
1288
1146-1166
AUGAAGUGGAUCUGGUAGGGAGA
1553
1144-1166





AD-569511.1
CCCUACCAGAUCCACUUCACU
1289
1147-1167
AGUGAAGUGGAUCUGGUAGGGAG
1554
1145-1167





AD-569512.1
CCUACCAGAUCCACUUCACCU
1290
1148-1168
AGGUGAAGUGGAUCUGGUAGGGA
1555
1146-1168





AD-569513.1
CUACCAGAUCCACUUCACCAU
1291
1149-1169
AUGGUGAAGUGGAUCUGGUAGGG
1556
1147-1169





AD-569514.1
UACCAGAUCCACUUCACCAAU
1292
1150-1170
AUUGGUGAAGUGGAUCUGGUAGG
1557
1148-1170





AD-569515.1
ACCAGAUCCACUUCACCAAGU
1293
1151-1171
ACUUGGUGAAGUGGAUCUGGUAG
1558
1149-1171





AD-569516.1
CCAGAUCCACUUCACCAAGAU
1294
1152-1172
AUCUUGGUGAAGUGGAUCUGGUA
1559
1150-1172





AD-569517.1
CAGAUCCACUUCACCAAGACU
1295
1153-1173
AGUCUUGGUGAAGUGGAUCUGGU
1560
1151-1173





AD-569518.1
AGAUCCACUUCACCAAGACAU
1296
1154-1174
AUGUCUUGGUGAAGUGGAUCUGG
1561
1152-1174





AD-569519.1
GAUCCACUUCACCAAGACACU
1297
1155-1175
AGUGUCUUGGUGAAGUGGAUCUG
1562
1153-1175





AD-569520.1
AUCCACUUCACCAAGACACCU
1298
1156-1176
AGGUGUCUUGGUGAAGUGGAUCU
1563
1154-1176





AD-569565.1
UUUGACCUCAUGGUGUUCGUU
1299
1201-1221
AACGAACACCAUGAGGUCAAAGG
1564
1199-1221





AD-569567.1
UGACCUCAUGGUGUUCGUGAU
1300
1203-1223
AUCACGAACACCAUGAGGUCAAA
1565
1201-1223





AD-570126.1
AGGGCGUGUUCGUGCUGAAUU
1301
1892-1912
AAUUCAGCACGAACACGCCCUUG
1566
1890-1912





AD-570127.1
GGGCGUGUUCGUGCUGAAUAU
1302
1893-1913
AUAUUCAGCACGAACACGCCCUU
1567
1891-1913





AD-570128.1
GGCGUGUUCGUGCUGAAUAAU
1303
1894-1914
AUUAUUCAGCACGAACACGCCCU
1568
1892-1914





AD-570129.1
GCGUGUUCGUGCUGAAUAAGU
1304
1895-1915
ACUUAUUCAGCACGAACACGCCC
1569
1893-1915





AD-570131.1
GUGUUCGUGCUGAAUAAGAAU
1305
1897-1917
AUUCUUAUUCAGCACGAACACGC
1570
1895-1917





AD-570135.1
UCGUGCUGAAUAAGAAGAACU
1306
1901-1921
AGUUCUUCUUAUUCAGCACGAAC
1571
1899-1921





AD-570136.1
CGUGCUGAAUAAGAAGAACAU
1307
1902-1922
AUGUUCUUCUUAUUCAGCACGAA
1572
1900-1922





AD-571535.1
AGACUUCCUUGAAGCCAACUU
1308
3612-3632
AAGUUGGCUUCAAGGAAGUCUCC
1573
3610-3632





AD-571536.1
GACUUCCUUGAAGCCAACUAU
1309
3613-3633
AUAGUUGGCUUCAAGGAAGUCUC
1574
3611-3633





AD-571537.1
ACUUCCUUGAAGCCAACUACU
1310
3614-3634
AGUAGUUGGCUUCAAGGAAGUCU
1575
3612-3634





AD-571538.1
CUUCCUUGAAGCCAACUACAU
1311
3615-3635
AUGUAGUUGGCUUCAAGGAAGUC
1576
3613-3635





AD-571540.1
UCCUUGAAGCCAACUACAUGU
1312
3617-3637
ACAUGUAGUUGGCUUCAAGGAAG
1577
3615-3637





AD-571541.1
CCUUGAAGCCAACUACAUGAU
1313
3618-3638
AUCAUGUAGUUGGCUUCAAGGAA
1578
3616-3638





AD-571542.1
CUUGAAGCCAACUACAUGAAU
1314
3619-3639
AUUCAUGUAGUUGGCUUCAAGGA
1579
3617-3639





AD-571543.1
UUGAAGCCAACUACAUGAACU
1315
3620-3640
AGUUCAUGUAGUUGGCUUCAAGG
1580
3618-3640





AD-571544.1
UGAAGCCAACUACAUGAACCU
1316
3621-3641
AGGUUCAUGUAGUUGGCUUCAAG
1581
3619-3641





AD-571545.1
GAAGCCAACUACAUGAACCUU
1317
3622-3642
AAGGUUCAUGUAGUUGGCUUCAA
1582
3620-3642





AD-571546.1
AAGCCAACUACAUGAACCUAU
1318
3623-3643
AUAGGUUCAUGUAGUUGGCUUCA
1583
3621-3643





AD-571547.1
AGCCAACUACAUGAACCUACU
1319
3624-3644
AGUAGGUUCAUGUAGUUGGCUUC
1584
3622-3644





AD-571548.1
GCCAACUACAUGAACCUACAU
1320
3625-3645
AUGUAGGUUCAUGUAGUUGGCUU
1585
3623-3645





AD-571549.1
CCAACUACAUGAACCUACAGU
1321
3626-3646
ACUGUAGGUUCAUGUAGUUGGCU
1586
3624-3646





AD-571550.1
CAACUACAUGAACCUACAGAU
1322
3627-3647
AUCUGUAGGUUCAUGUAGUUGGC
1587
3625-3647





AD-571551.1
AACUACAUGAACCUACAGAGU
1323
3628-3648
ACUCUGUAGGUUCAUGUAGUUGG
1588
3626-3648





AD-571552.1
ACUACAUGAACCUACAGAGAU
1324
3629-3649
AUCUCUGUAGGUUCAUGUAGUUG
1589
3627-3649





AD-571553.1
CUACAUGAACCUACAGAGAUU
1325
3630-3650
AAUCUCUGUAGGUUCAUGUAGUU
1590
3628-3650





AD-571554.1
UACAUGAACCUACAGAGAUCU
1326
3631-3651
AGAUCUCUGUAGGUUCAUGUAGU
1591
3629-3651





AD-571555.1
ACAUGAACCUACAGAGAUCCU
1327
3632-3652
AGGAUCUCUGUAGGUUCAUGUAG
1592
3630-3652





AD-571556.1
CAUGAACCUACAGAGAUCCUU
1328
3633-3653
AAGGAUCUCUGUAGGUUCAUGUA
1593
3631-3653





AD-571557.1
AUGAACCUACAGAGAUCCUAU
1329
3634-3654
AUAGGAUCUCUGUAGGUUCAUGU
1594
3632-3654





AD-571558.1
UGAACCUACAGAGAUCCUACU
1330
3635-3655
AGUAGGAUCUCUGUAGGUUCAUG
1595
3633-3655





AD-571559.1
GAACCUACAGAGAUCCUACAU
1331
3636-3656
AUGUAGGAUCUCUGUAGGUUCAU
1596
3634-3656





AD-571560.1
AACCUACAGAGAUCCUACACU
1332
3637-3657
AGUGUAGGAUCUCUGUAGGUUCA
1597
3635-3657





AD-571711.1
GGCCCUACUGCAGCUAAAAGU
1333
3807-3827
ACUUUUAGCUGCAGUAGGGCCAA
1598
3805-3827





AD-571712.1
GCCCUACUGCAGCUAAAAGAU
1334
3808-3828
AUCUUUUAGCUGCAGUAGGGCCA
1599
3806-3828





AD-571713.1
CCCUACUGCAGCUAAAAGACU
1335
3809-3829
AGUCUUUUAGCUGCAGUAGGGCC
1600
3807-3829





AD-571714.1
CCUACUGCAGCUAAAAGACUU
1336
3810-3830
AAGUCUUUUAGCUGCAGUAGGGC
1601
3808-3830





AD-571716.1
UACUGCAGCUAAAAGACUUUU
1337
3812-3832
AAAAGUCUUUUAGCUGCAGUAGG
1602
3810-3832





AD-571717.1
ACUGCAGCUAAAAGACUUUGU
1338
3813-3833
ACAAAGUCUUUUAGCUGCAGUAG
1603
3811-3833





AD-571718.1
CUGCAGCUAAAAGACUUUGAU
1339
3814-3834
AUCAAAGUCUUUUAGCUGCAGUA
1604
3812-3834





AD-571719.2
UGCAGCUAAAAGACUUUGACU
1340
3815-3835
AGUCAAAGUCUUUUAGCUGCAGU
1605
3813-3835





AD-571720.1
GCAGCUAAAAGACUUUGACUU
1341
3816-3836
AAGUCAAAGUCUUUUAGCUGCAG
1606
3814-3836





AD-571721.1
CAGCUAAAAGACUUUGACUUU
1342
3817-3837
AAAGUCAAAGUCUUUUAGCUGCA
1607
3815-3837





AD-571722.1
AGCUAAAAGACUUUGACUUUU
1343
3818-3838
AAAAGUCAAAGUCUUUUAGCUGC
1608
3816-3838





AD-571723.1
GCUAAAAGACUUUGACUUUGU
1344
3819-3839
ACAAAGUCAAAGUCUUUUAGCUG
1609
3817-3839





AD-571742.1
GUGCCUCCCGUCGUGCGUUGU
1345
3838-3858
ACAACGCACGACGGGAGGCACAA
1610
3836-3858





AD-571743.1
UGCCUCCCGUCGUGCGUUGGU
1346
3839-3859
ACCAACGCACGACGGGAGGCACA
1611
3837-3859





AD-571744.1
GCCUCCCGUCGUGCGUUGGCU
1347
3840-3860
AGCCAACGCACGACGGGAGGCAC
1612
3838-3860





AD-571745.1
CCUCCCGUCGUGCGUUGGCUU
1348
3841-3861
AAGCCAACGCACGACGGGAGGCA
1613
3839-3861





AD-571746.1
CUCCCGUCGUGCGUUGGCUCU
1349
3842-3862
AGAGCCAACGCACGACGGGAGGC
1614
3840-3862





AD-571747.1
UCCCGUCGUGCGUUGGCUCAU
1350
3843-3863
AUGAGCCAACGCACGACGGGAGG
1615
3841-3863





AD-571748.1
CCCGUCGUGCGUUGGCUCAAU
1351
3844-3864
AUUGAGCCAACGCACGACGGGAG
1616
3842-3864





AD-571749.1
CCGUCGUGCGUUGGCUCAAUU
1352
3845-3865
AAUUGAGCCAACGCACGACGGGA
1617
3843-3865





AD-571750.1
CGUCGUGCGUUGGCUCAAUGU
1353
3846-3866
ACAUUGAGCCAACGCACGACGGG
1618
3844-3866





AD-571751.1
GUCGUGCGUUGGCUCAAUGAU
1354
3847-3867
AUCAUUGAGCCAACGCACGACGG
1619
3845-3867





AD-571753.2
CGUGCGUUGGCUCAAUGAACU
1355
3849-3869
AGUUCAUUGAGCCAACGCACGAC
1620
3847-3869





AD-571755.1
UGCGUUGGCUCAAUGAACAGU
1356
3851-3871
ACUGUUCAUUGAGCCAACGCACG
1621
3849-3871





AD-571756.1
GCGUUGGCUCAAUGAACAGAU
1357
3852-3872
AUCUGUUCAUUGAGCCAACGCAC
1622
3850-3872





AD-571757.1
CGUUGGCUCAAUGAACAGAGU
1358
3853-3873
ACUCUGUUCAUUGAGCCAACGCA
1623
3851-3873





AD-571758.1
GUUGGCUCAAUGAACAGAGAU
1359
3854-3874
AUCUCUGUUCAUUGAGCCAACGC
1624
3852-3874





AD-571759.1
UUGGCUCAAUGAACAGAGAUU
1360
3855-3875
AAUCUCUGUUCAUUGAGCCAACG
1625
3853-3875





AD-571760.1
UGGCUCAAUGAACAGAGAUAU
1361
3856-3876
AUAUCUCUGUUCAUUGAGCCAAC
1626
3854-3876





AD-571761.1
GGCUCAAUGAACAGAGAUACU
1362
3857-3877
AGUAUCUCUGUUCAUUGAGCCAA
1627
3855-3877





AD-571762.1
GCUCAAUGAACAGAGAUACUU
1363
3858-3878
AAGUAUCUCUGUUCAUUGAGCCA
1628
3856-3878





AD-571763.1
CUCAAUGAACAGAGAUACUAU
1364
3859-3879
AUAGUAUCUCUGUUCAUUGAGCC
1629
3857-3879





AD-571764.1
UCAAUGAACAGAGAUACUACU
1365
3860-3880
AGUAGUAUCUCUGUUCAUUGAGC
1630
3858-3880





AD-571765.2
CAAUGAACAGAGAUACUACGU
1366
3861-3881
ACGUAGUAUCUCUGUUCAUUGAG
1631
3859-3881





AD-571766.2
AAUGAACAGAGAUACUACGGU
1367
3862-3882
ACCGUAGUAUCUCUGUUCAUUGA
1632
3860-3882





AD-571767.2
AUGAACAGAGAUACUACGGUU
1368
3863-3883
AACCGUAGUAUCUCUGUUCAUUG
1633
3861-3883





AD-572383.1
GCAGUCAAGGUCUACGCCUAU
1369
4519-4539
AUAGGCGUAGACCUUGACUGCUC
1634
4517-4539





AD-572384.1
CAGUCAAGGUCUACGCCUAUU
1370
4520-4540
AAUAGGCGUAGACCUUGACUGCU
1635
4518-4540





AD-572385.1
AGUCAAGGUCUACGCCUAUUU
1371
4521-4541
AAAUAGGCGUAGACCUUGACUGC
1636
4519-4541





AD-572386.1
GUCAAGGUCUACGCCUAUUAU
1372
4522-4542
AUAAUAGGCGUAGACCUUGACUG
1637
4520-4542





AD-572387.4
UCAAGGUCUACGCCUAUUACU
1373
4523-4543
AGUAAUAGGCGUAGACCUUGACU
1638
4521-4543





AD-572391.1
GGUCUACGCCUAUUACAACCU
1374
4527-4547
AGGUUGUAAUAGGCGUAGACCUU
1639
4525-4547





AD-572392.1
GUCUACGCCUAUUACAACCUU
1375
4528-4548
AAGGUUGUAAUAGGCGUAGACCU
1640
4526-4548





AD-572393.2
UCUACGCCUAUUACAACCUGU
1376
4529-4549
ACAGGUUGUAAUAGGCGUAGACC
1641
4527-4549





AD-572394.1
CUACGCCUAUUACAACCUGGU
1377
4530-4550
ACCAGGUUGUAAUAGGCGUAGAC
1642
4528-4550





AD-572395.1
UACGCCUAUUACAACCUGGAU
1378
4531-4551
AUCCAGGUUGUAAUAGGCGUAGA
1643
4529-4551





AD-572396.1
ACGCCUAUUACAACCUGGAGU
1379
4532-4552
ACUCCAGGUUGUAAUAGGCGUAG
1644
4530-4552





AD-572397.1
CGCCUAUUACAACCUGGAGGU
1380
4533-4553
ACCUCCAGGUUGUAAUAGGCGUA
1645
4531-4553





AD-572495.1
GCUGAGGAGAAUUGCUUCAUU
1381
4633-4653
AAUGAAGCAAUUCUCCUCAGCAC
1646
4631-4653





AD-572569.1
GCCAGGAGUGGACUAUGUGUU
1382
4707-4727
AACACAUAGUCCACUCCUGGCUC
1647
4705-4727





AD-572570.1
CCAGGAGUGGACUAUGUGUAU
1383
4708-4728
AUACACAUAGUCCACUCCUGGCU
1648
4706-4728





AD-572571.1
CAGGAGUGGACUAUGUGUACU
1384
4709-4729
AGUACACAUAGUCCACUCCUGGC
1649
4707-4729





AD-572572.1
AGGAGUGGACUAUGUGUACAU
1385
4710-4730
AUGUACACAUAGUCCACUCCUGG
1650
4708-4730





AD-572573.1
GGAGUGGACUAUGUGUACAAU
1386
4711-4731
AUUGUACACAUAGUCCACUCCUG
1651
4709-4731





AD-572574.1
GAGUGGACUAUGUGUACAAGU
1387
4712-4732
ACUUGUACACAUAGUCCACUCCU
1652
4710-4732





AD-572575.1
AGUGGACUAUGUGUACAAGAU
1388
4713-4733
AUCUUGUACACAUAGUCCACUCC
1653
4711-4733





AD-572576.1
GUGGACUAUGUGUACAAGACU
1389
4714-4734
AGUCUUGUACACAUAGUCCACUC
1654
4712-4734





AD-572577.1
UGGACUAUGUGUACAAGACCU
1390
4715-4735
AGGUCUUGUACACAUAGUCCACU
1655
4713-4735





AD-572580.1
ACUAUGUGUACAAGACCCGAU
1391
4718-4738
AUCGGGUCUUGUACACAUAGUCC
1656
4716-4738





AD-572581.1
CUAUGUGUACAAGACCCGACU
1392
4719-4739
AGUCGGGUCUUGUACACAUAGUC
1657
4717-4739
















TABLE 21







Modified Sense and Antisense Strand Sequences


of Complement Component C3 dsRNA Agents
















Anti-

mRNA




Sense

sense

Target




Se-
SEQ
Se-
SEQ
Se-
SEQ


Duplex
quence
ID
quence
ID
quence
ID


Name
5′ to 3′
NO:
5′ to 3′
NO:
5′ to 3′
NO:





AD-
gsusgc
1658
asUfsu
1923
UCGUGC
2188


570137.1
ugAfaU

guUfcU

UGAAUA




fAfAfg

fUfcuu

AGAAGA




aagaac

aUfuCf

ACAAA




aauL96

agcacs








gsa








AD-
usgscu
1659
asUfsu
1924
CGUGCU
2189


570138.1
gaAfuA

ugUfuC

GAAUAA




fAfGfa

fUfucu

GAAGAA




agaaca

uAfuUf

CAAAC




aauL96

cagcas








csg








AD-
gscsug
1660
asGfsu
1925
GUGCUG
2190


570139.1
aaUfaA

uuGfuU

AAUAAG




fGfAfa

fCfuuc

AAGAAC




gaacaa

uUfaUf

AAACU




acuL96

ucagcs








asc








AD-
csusga
1661
asAfsg
1926
UGCUGA
2191


570140.1
auAfaG

uuUfgU

AUAAGA




fAfAfg

fUfcuu

AGAACA




aacaaa

cUfuAf

AACUG




cuuL96

uucags








csa








AD-
usgsaa
1662
asCfsa
1927
GCUGAA
2192


570141.1
uaAfgA

guUfuG

UAAGAA




fAfGfa

fUfucu

GAACAA




acaaac

uCfuUf

ACUGA




uguL96

auucas








gsc








AD-
gsasau
1663
asUfsc
1928
CUGAAU
2193


570142.1
aaGfaA

agUfuU

AAGAAG




fGfAfa

fGfuuc

AACAAA




caaacu

uUfcUf

CUGAC




gauL96

uauucs








asg








AD-
asasua
1664
asGfsu
1929
UGAAUA
2194


570143.1
agAfaG

caGfuU

AGAAGA




fAfAfc

fUfguu

ACAAAC




aaacug

cUfuCf

UGACG




acuL96

uuauus








csa








AD-
asusaa
1665
asCfsg
1930
GAAUAA
2195


570144.1
gaAfgA

ucAfgU

GAAGAA




fAfCfa

fUfugu

CAAACU




aacuga

uCfuUf

GACGC




cguL96

cuuaus








usc








AD-
usasag
1666
asGfsc
1931
AAUAAG
2196


570145.1
aaGfaA

guCfaG

AAGAAC




fCfAfa

fUfuug

AAACUG




acugac

uUfcUf

ACGCA




gcuL96

ucuuas








usu








AD-
asasga
1667
asUfsg
1932
AUAAGA
2197


570146.1
agAfaC

cgUfcA

AGAACA




fAfAfa

fGfuuu

AACUGA




cugacg

gUfuCf

CGCAG




cauL96

uucuus








asu








AD-
asgsaa
1668
asCfsu
1933
UAAGAA
2198


570147.1
gaAfcA

gcGfuC

GAACAA




fAfAfc

fAfguu

ACUGAC




ugacgc

uGfuUf

GCAGA




aguL96

cuucus








usa








AD-
gsasag
1669
asUfsc
1934
AAGAAG
2199


570148.1
aaCfaA

ugCfgU

AACAAA




fAfCfu

fCfagu

CUGACG




gacgca

uUfgUf

CAGAG




gauL96

ucuucs








usu








AD-
asasga
1670
asCfsu
1935
AGAAGA
2200


570149.1
acAfaA

cuGfcG

ACAAAC




fCfUfg

fUfcag

UGACGC




acgcag

uUfuGf

AGAGU




aguL96

uucuus








csu








AD-
asgsaa
1671
asAfsc
1936
GAAGAA
2201


570150.1
caAfaC

ucUfgC

CAAACU




fUfGfa

fGfuca

GACGCA




cgcaga

gUfuUf

GAGUA




guuL96

guucus








usc








AD-
gsasac
1672
asUfsa
1937
AAGAAC
2202


570151.1
aaAfcU

cuCfuG

AAACUG




fGfAfg

fCfguc

ACGCAG




cagagu

aGfuUf

AGUAA




auL96

uguucs








usu








AD-
asasca
1673
asUfsu
1938
AGAACA
2203


570152.1
aaCfuG

acUfcU

AACUGA




fAfCfg

fGfcgu

CGCAGA




cagagu

cAfgUf

GUAAG




aauL96

uuguus








csu








AD-
ascsaa
1674
asCfsu
1939
GAACAA
2204


570153.1
acUfgA

uaCfuC

ACUGAC




fCfGfc

fUfgcg

GCAGAG




agagua

uCfaGf

UAAGA




aguL96

uuugus








usc








AD-
csasaa
1675
asUfsc
1940
AACAAA
2205


570154.1
cuGfaC

uuAfcU

CUGACG




fGfCfa

fCfugc

CAGAGU




gaguaa

gUfcAf

AAGAU




gauL96

guuugs








usu








AD-
asasac
1676
asAfsu
1941
ACAAAC
2206


570155.1
ugAfcG

cuUfaC

UGACGC




fCfAfg

fUfcug

AGAGUA




aguaag

cGfuCf

AGAUC




auuL96

aguuus








gsu








AD-
asascu
1677
asGfsa
1942
CAAACU
2207


570156.2
gaCfgC

ucUfuA

GACGCA




fAfGfa

fCfucu

GAGUAA




guaaga

gCfgUf

GAUCU




ucuL96

caguus








usg








AD-
csusga
1678
asCfsa
1943
AACUGA
2208


570158.1
cgCfaG

gaUfcU

CGCAGA




fAfGfu

fUfacu

GUAAGA




aagauc

cUfgCf

UCUGG




uguL96

gucags








usu








AD-
usgsac
1679
asCfsc
1944
ACUGAC
2209


570159.1
gcAfgA

agAfuC

GCAGAG




fGfUfa

fUfuac

UAAGAU




agaucu

uCfuGf

CUGGG




gguL96

cgucas








gsu








AD-
gsascg
1680
asCfsc
1945
CUGACG
2210


570160.1
caGfaG

caGfaU

CAGAGU




fUfAfa

fCfuua

AAGAUC




gaucug

cUfcUf

UGGGA




gguL96

gcgucs








asg








AD-
ascsgc
1681
asUfsc
1946
UGACGC
2211


570161.1
agAfgU

ccAfgA

AGAGUA




fAfAfg

fUfcuu

AGAUCU




aucugg

aCfuCf

GGGAC




gauL96

ugcgus








csa








AD-
usgsag
1682
asUfsu
1947
UGUGAG
2212


570611.1
caUfgU

ucUfuG

CAUGUC




fCfGfg

fUfccg

GGACAA




acaaga

aCfaUf

GAAAG




aauL96

gcucas








csa








AD-
gsasgc
1683
asCfsu
1948
GUGAGC
2213


570612.1
auGfuC

uuCfuU

AUGUCG




fGfGfa

fGfucc

GACAAG




caagaa

gAfcAf

AAAGG




aguL96

ugcucs








asc








AD-
asgsca
1684
asCfsc
1949
UGAGCA
2214


570613.1
ugUfcG

uuUfcU

UGUCGG




fGfAfc

fUfguc

ACAAGA




aagaaa

cGfaCf

AAGGG




gguL96

augcus








csa








AD-
gscsau
1685
asCfsc
1950
GAGCAU
2215


570614.1
guCfgG

cuUfuC

GUCGGA




fAfCfa

fUfugu

CAAGAA




agaaag

cCfgAf

AGGGA




gguL96

caugcs








usc








AD-
csasug
1686
asUfsc
1951
AGCAUG
2216


570615.1
ucGfgA

ccUfuU

UCGGAC




fCfAfa

fCfuug

AAGAAA




gaaagg

uCfcGf

GGGAU




gauL96

acaugs








csu








AD-
asusgu
1687
asAfsu
1952
GCAUGU
2217


570616.1
cgGfaC

ccCfuU

CGGACA




fAfAfg

fUfcuu

AGAAAG




aaaggg

gUfcCf

GGAUC




auuL96

gacaus








gsc








AD-
usgsuc
1688
asGfsa
1953
CAUGUC
2218


570617.1
ggAfcA

ucCfcU

GGACAA




fAfGfa

fUfucu

GAAAGG




aaggga

uGfuCf

GAUCU




ucuL96

egacas








usg








AD-
gsuscg
1689
asAfsg
1954
AUGUCG
2219


570618.1
gaCfaA

auCfcC

GACAAG




fGfAfa

fUfuuc

AAAGGG




agggau

uUfgUf

AUCUG




cuuL96

ccgacs








asu








AD-
uscsgg
1690
asCfsa
1955
UGUCGG
2220


570619.1
acAfaG

gaUfcC

ACAAGA




fAfAfa

fCfuuu

AAGGGA




gggauc

cUfuGf

UCUGU




uguL96

uccgas








csa








AD-
csgsga
1691
asAfsc
1956
GUCGGA
2221


570620.3
caAfgA

agAfuC

CAAGAA




fAfAfg

fCfcuu

AGGGAU




ggaucu

uCfuUf

CUGUG




guuL96

guccgs








asc








AD-
gsgsac
1692
asCfsa
1957
UCGGAC
2222


570621.2
aaGfaA

caGfaU

AAGAAA




fAfGfg

fCfccu

GGGAUC




gaucug

uUfcUf

UGUGU




uguL96

uguccs








gsa








AD-
gsasca
1693
asAfsc
1958
CGGACA
2223


570622.2
agAfaA

acAfgA

AGAAAG




fGfGfg

fUfccc

GGAUCU




aucugu

uUfuCf

GUGUG




guuL96

uugucs








csg








AD-
ascsaa
1694
asCfsa
1959
GGACAA
2224


570623.4
gaAfaG

caCfaG

GAAAGG




fGfGfa

fAfucc

GAUCUG




ucugug

cUfuUf

UGUGG




uguL96

cuugus








csc








AD-
csasag
1695
asCfsc
1960
GACAAG
2225


570624.2
aaAfgG

acAfcA

AAAGGG




fGfAfu

fGfauc

AUCUGU




cugugu

cCfuUf

GUGGC




gguL96

ucuugs








usc








AD-
asasga
1696
asGfsc
1961
ACAAGA
2226


570625.2
aaGfgG

caCfaC

AAGGGA




fAfUfc

fAfgau

UCUGUG




ugugug

cCfcUf

UGGCA




gcuL96

uucuus








gsu








AD-
asgsaa
1697
asUfsg
1962
CAAGAA
2227


570626.1
agGfgA

ccAfcA

AGGGAU




fUfCfu

fCfaga

CUGUGU




gugugg

uCfcCf

GGCAG




cauL96

uuucus








usg








AD-
gsasaa
1698
asCfsu
1963
AAGAAA
2228


570627.2
ggGfaU

gcCfaC

GGGAUC




fCfUfg

fAfcag

UGUGUG




uguggc

aUfcCf

GCAGA




aguL96

cuuucs








usu








AD-
asasag
1699
asUfsc
1964
AGAAAG
2229


570628.1
ggAfuC

ugCfcA

GGAUCU




fUfGfu

fCfaca

GUGUGG




guggca

gAfuCf

CAGAC




gauL96

ccuuus








csu








AD-
asasgg
1700
asGfsu
1965
GAAAGG
2230


570629.1
gaUfcU

cuGfcC

GAUCUG




fGfUfg

fAfcac

UGUGGC




uggcag

aGfaUf

AGACC




acuL96

cccuus








usc








AD-
asgsgg
1701
asGfsg
1966
AAAGGG
2231


570630.1
auCfuG

ucUfgC

AUCUGU




fUfGfu

fCfaca

GUGGCA




ggcaga

cAfgAf

GACCC




ccuL96

ucccus








usu








AD-
gsgsga
1702
asGfsg
1967
AAGGGA
2232


1069837.1
ucUfgU

guCfuG

UCUGUG




fGfUfg

fCfcac

UGGCAG




gcagac

aCfaGf

ACCCC




ccuL96

aucccs








usu








AD-
gsasaa
1703
asUfsa
1968
UGGAAA
2233


570707.1
ucCfgA

gaGfaA

UCCGAG




fGfCfc

fCfggc

CCGUUC




guucuc

uCfgGf

UCUAC




uauL96

auuucs








csa








AD-
asasau
1704
asGfsu
1969
GGAAAU
2234


570708.1
ccGfaG

agAfgA

CCGAGC




fCfCfg

fAfcgg

CGUUCU




uucucu

cUfcGf

CUACA




acuL96

gauuus








csc








AD-
asasuc
1705
asUfsg
1970
GAAAUC
2235


570709.1
cgAfgC

uaGfaG

CGAGCC




fCfGfu

fAfacg

GUUCUC




ucucua

gCfuCf

UACAA




cauL96

ggauus








usc








AD-
asuscc
1706
asUfsu
1971
AAAUCC
2236


570710.1
gaGfcC

guAfgA

GAGCCG




fGfUfu

fGfaac

UUCUCU




cucuac

gGfcUf

ACAAU




aauL96

cggaus








usu








AD-
asgscc
1707
asGfsg
1972
CGAGCC
2237


570715.1
guUfcU

uaAfuU

GUUCUC




fCfUfa

fGfuag

UACAAU




caauua

aGfaAf

UACCG




ccuL96

cggcus








csg








AD-
gscscg
1708
asCfsg
1973
GAGCCG
2238


570716.1
uuCfuC

guAfaU

UUCUCU




fUfAfc

fUfgua

ACAAUU




aauuac

gAfgAf

ACCGG




cguL96

acggcs








usc








AD-
cscsgu
1709
asCfsc
1974
AGCCGU
2239


570717.2
ucUfcU

ggUfaA

UCUCUA




fAfCfa

fUfugu

CAAUUA




auuacc

aGfaGf

CCGGC




gguL96

aacggs








csu








AD-
csgsuu
1710
asGfsc
1975
GCCGUU
2240


570718.1
cuCfuA

cgGfuA

CUCUAC




fCfAfa

fAfuug

AAUUAC




uuaccg

uAfgAf

CGGCA




gcuL96

gaacgs








gsc








AD-
gsusuc
1711
asUfsg
1976
CCGUUC
2241


570719.1
ucUfaC

ccGfgU

UCUACA




fAfAfu

fAfauu

AUUACC




uaccgg

gUfaGf

GGCAG




cauL96

agaacs








gsg








AD-
ususcu
1712
asCfsu
1977
CGUUCU
2242


570720.1
cuAfcA

gcCfgG

CUACAA




fAfUfu

fUfaau

UUACCG




accggc

uGfuAf

GCAGA




aguL96

gagaas








csg








AD-
uscsuc
1713
asUfsc
1978
GUUCUC
2243


570721.1
uaCfaA

ugCfcG

UACAAU




fUfUfa

fGfuaa

UACCGG




ccggca

uUfgUf

CAGAA




gauL96

agagas








asc








AD-
gsgscu
1714
asGfsa
1979
CUGGCU
2244


571285.1
gaCfcG

ccAfcG

GACCGC




fCfCfu

fUfagg

CUACGU




acgugg

cGfgUf

GGUCA




ucuL96

cagccs








asg








AD-
gscsug
1715
asUfsg
1980
UGGCUG
2245


571286.1
acCfgC

acCfaC

ACCGCC




fCfUfa

fGfuag

UACGUG




cguggu

gCfgGf

GUCAA




cauL96

ucagcs








csa








AD-
csusga
1716
asUfsu
1981
GGCUGA
2246


571287.1
ccGfcC

gaCfcA

CCGCCU




fUfAfc

fCfgua

ACGUGG




gugguc

gGfcGf

UCAAG




aauL96

gucags








csc








AD-
usgsac
1717
asCfsu
1982
GCUGAC
2247


571288.1
cgCfcU

ugAfcC

CGCCUA




fAfCfg

fAfcgu

CGUGGU




ugguca

aGfgCf

CAAGG




aguL96

ggucas








gsc








AD-
gsascc
1718
asCfsc
1983
CUGACC
2248


571289.1
gcCfuA

uuGfaC

GCCUAC




fCfGfu

fCfacg

GUGGUC




ggucaa

uAfgGf

AAGGU




gguL96

cggucs








asg








AD-
ascscg
1719
asAfsc
1984
UGACCG
2249


571290.1
ccUfaC

cuUfgA

CCUACG




fGfUfg

fCfcac

UGGUCA




gucaag

gUfaGf

AGGUC




guuL96

gcggus








csa








AD-
cscsgc
1720
asGfsa
1985
GACCGC
2250


571291.1
cuAfcG

ccUfuG

CUACGU




fUfGfg

fAfcca

GGUCAA




ucaagg

cGfuAf

GGUCU




ucuL96

ggcggs








usc








AD-
csgscc
1721
asAfsg
1986
ACCGCC
2251


571292.1
uaCfgU

acCfuU

UACGUG




fGfGfu

fGfacc

GUCAAG




caaggu

aCfgUf

GUCUU




cuuL96

aggcgs








gsu








AD-
gscscu
1722
asAfsa
1987
CCGCCU
2252


571293.1
acGfuG

gaCfcU

ACGUGG




fGfUfc

fUfgac

UCAAGG




aagguc

cAfcGf

UCUUC




uuuL96

uaggcs








gsg








AD-
cscsua
1723
asGfsa
1988
CGCCUA
2253


571294.1
cgUfgG

agAfcC

CGUGGU




fUfCfa

fUfuga

CAAGGU




aggucu

cCfaCf

CUUCU




ucuL96

guaggs








csg








AD-
csusac
1724
asAfsg
1989
GCCUAC
2254


571295.1
guGfgU

aaGfaC

GUGGUC




fCfAfa

fCfuug

AAGGUC




ggucuu

aCfcAf

UUCUC




cuuL96

cguags








gsc








AD-
usascg
1725
asGfsa
1990
CCUACG
2255


571296.1
ugGfuC

gaAfgA

UGGUCA




fAfAfg

fCfcuu

AGGUCU




gucuuc

gAfcCf

UCUCU




ucuL96

acguas








gsg








AD-
ascsgu
1726
asAfsg
1991
CUACGU
2256


571297.1
ggUfcA

agAfaG

GGUCAA




fAfGfg

fAfccu

GGUCUU




ucuucu

uGfaCf

CUCUC




cuuL96

cacgus








asg








AD-
csgsug
1727
asGfsa
1992
UACGUG
2257


571298.6
guCfaA

gaGfaA

GUCAAG




fGfGfu

fGfacc

GUCUUC




cuucuc

uUfgAf

UCUCU




ucuL96

ccacgs








usa








AD-
gsusgg
1728
asAfsg
1993
ACGUGG
2258


571299.1
ucAfaG

agAfgA

UCAAGG




fGfUfc

fAfgac

UCUUCU




uucucu

cUfuGf

CUCUG




cuuL96

accacs








gsu








AD-
usgsgu
1729
asCfsa
1994
CGUGGU
2259


571300.1
caAfgG

gaGfaG

CAAGGU




fUfCfu

fAfaga

CUUCUC




ucucuc

cCfuUf

UCUGG




uguL96

gaccas








csg








AD-
gsgsuc
1730
asCfsc
1995
GUGGUC
2260


571301.1
aaGfgU

agAfgA

AAGGUC




fCfUfu

fGfaag

UUCUCU




cucucu

aCfcUf

CUGGC




gguL96

ugaccs








asc








AD-
gsusca
1731
asGfsc
1996
UGGUCA
2261


571302.1
agGfuC

caGfaG

AGGUCU




fUfUfc

fAfgaa

UCUCUC




ucucug

gAfcCf

UGGCU




gcuL96

uugacs








csa








AD-
uscsaa
1732
asAfsg
1997
GGUCAA
2262


571303.1
ggUfcU

ccAfgA

GGUCUU




fUfCfu

fGfaga

CUCUCU




cucugg

aGfaCf

GGCUG




cuuL96

cuugas








csc








AD-
csasag
1733
asCfsa
1998
GUCAAG
2263


571304.1
guCfuU

gcCfaG

GUCUUC




fCfUfc

fAfgag

UCUCUG




ucuggc

aAfgAf

GCUGU




uguL96

ccuugs








asc








AD-
asasgg
1734
asAfsc
1999
UCAAGG
2264


571305.1
ucUfuC

agCfcA

UCUUCU




fUfCfu

fGfaga

CUCUGG




cuggcu

gAfaGf

CUGUC




guuL96

accuus








gsa








AD-
asgsgu
1735
asGfsa
2000
CAAGGU
2265


571306.1
cuUfcU

caGfcC

CUUCUC




fCfUfc

fAfgag

UCUGGC




uggcug

aGfaAf

UGUCA




ucuL96

gaccus








usg








AD-
gsgsuc
1736
asUfsg
2001
AAGGUC
2266


571307.1
uuCfuC

acAfgC

UUCUCU




fUfCfu

fCfaga

CUGGCU




ggcugu

gAfgAf

GUCAA




cauL96

agaccs








usu








AD-
gsuscu
1737
asUfsu
2002
AGGUCU
2267


571308.1
ucUfcU

gaCfaG

UCUCUC




fCfUfg

fCfcag

UGGCUG




gcuguc

aGfaGf

UCAAC




aauL96

aagacs








csu








AD-
uscsuu
1738
asGfsu
2003
GGUCUU
2268


571309.1
cuCfuC

ugAfcA

CUCUCU




fUfGfg

fGfcca

GGCUGU




cuguca

gAfgAf

CAACC




acuL96

gaagas








csc








AD-
usasaa
1739
asCfsa
2004
ACUAAA
2269


571526.1
gcAfgG

agGfaA

GCAGGA




fAfGfa

fGfucu

GACUUC




cuuccu

cCfuGf

CUUGA




uguL96

cuuuas








gsu








AD-
asasag
1740
asUfsc
2005
CUAAAG
2270


571527.1
caGfgA

aaGfgA

CAGGAG




fGfAfc

fAfguc

ACUUCC




uuccuu

uCfcUf

UUGAA




gauL96

gcuuus








asg








AD-
asasgc
1741
asUfsu
2006
UAAAGC
2271


571528.1
agGfaG

caAfgG

AGGAGA




fAfCfu

fAfagu

CUUCCU




uccuug

cUfcCf

UGAAG




aauL96

ugcuus








usa








AD-
asgsca
1742
asCfsu
2007
AAAGCA
2272


571529.1
ggAfgA

ucAfaG

GGAGAC




fCfUfu

fGfaag

UUCCUU




ccuuga

uCfuCf

GAAGC




aguL96

cugcus








usu








AD-
gscsag
1743
asGfsc
2008
AAGCAG
2273


571530.1
gaGfaC

uuCfaA

GAGACU




fUfUfc

fGfgaa

UCCUUG




cuugaa

gUfcUf

AAGCC




gcuL96

ccugcs








usu








AD-
csasgg
1744
asGfsg
2009
AGCAGG
2274


571531.1
agAfcU

cuUfcA

AGACUU




fUfCfc

fAfgga

CCUUGA




uugaag

aGfuCf

AGCCA




ccuL96

uccugs








csu








AD-
asgsga
1745
asUfsg
2010
GCAGGA
2275


571532.1
gaCfuU

gcUfuC

GACUUC




fCfCfu

fAfagg

CUUGAA




ugaagc

aAfgUf

GCCAA




cauL96

cuccus








gsc








AD-
gsgsag
1746
asUfsu
2011
CAGGAG
2276


571533.1
acUfuC

ggCfuU

ACUUCC




fCfUfu

fCfaag

UUGAAG




gaagcc

gAfaGf

CCAAC




aauL96

ucuccs








usg








AD-
gsasga
1747
asGfsu
2012
AGGAGA
2277


571534.1
cuUfcC

ugGfcU

CUUCCU




fUfUfg

fUfcaa

UGAAGC




aagcca

gGfaAf

CAACU




acuL96

gucucs








csu








AD-
asgsag
1748
asGfsa
2013
GCAGAG
2278


568955.1
cgGfgU

ugAfaG

CGGGUA




fAfCfc

fAfggu

CCUCUU




ucuuca

aCfcCf

CAUCC




ucuL96

gcucus








gsc








AD-
gsasgc
1749
asGfsg
2014
CAGAGC
2279


568956.1
ggGfuA

auGfaA

GGGUAC




fCfCfu

fGfagg

CUCUUC




cuucau

uAfcCf

AUCCA




ccuL96

cgcucs








usg








AD-
asgscg
1750
asUfsg
2015
AGAGCG
2280


568957.1
ggUfaC

gaUfgA

GGUACC




fCfUfc

fAfgag

UCUUCA




uucauc

gUfaCf

UCCAG




cauL96

ccgcus








csu








AD-
gscsgg
1751
asCfsu
2016
GAGCGG
2281


568958.1
guAfcC

ggAfuG

GUACCU




fUfCfu

fAfaga

CUUCAU




ucaucc

gGfuAf

CCAGA




aguL96

cccgcs








usc








AD-
csgsgg
1752
asUfsc
2017
AGCGGG
2282


568959.1
uaCfcU

ugGfaU

UACCUC




fCfUfu

fGfaag

UUCAUC




caucca

aGfgUf

CAGAC




gauL96

acccgs








csu








AD-
gsgsgu
1753
asGfsu
2018
GCGGGU
2283


568960.1
acCfuC

cuGfgA

ACCUCU




fUfUfc

fUfgaa

UCAUCC




auccag

gAfgGf

AGACA




acuL96

uacccs








gsc








AD-
gsgsua
1754
asUfsg
2019
CGGGUA
2284


568961.1
ccUfcU

ucUfgG

CCUCUU




fUfCfa

fAfuga

CAUCCA




uccaga

aGfaGf

GACAG




cauL96

guaccs








csg








AD-
gsusac
1755
asCfsu
2020
GGGUAC
2285


568962.1
cuCfuU

guCfuG

CUCUUC




fCfAfu

fGfaug

AUCCAG




ccagac

aAfgAf

ACAGA




aguL96

gguacs








csc








AD-
usascc
1756
asUfsc
2021
GGUACC
2286


568963.2
ucUfuC

ugUfcU

UCUUCA




fAfUfc

fGfgau

UCCAGA




cagaca

gAfaGf

CAGAC




gauL96

agguas








csc








AD-
ascscu
1757
asGfsu
2022
GUACCU
2287


568964.1
cuUfcA

cuGfuC

CUUCAU




fUfCfc

fUfgga

CCAGAC




agacag

uGfaAf

AGACA




acuL96

gaggus








asc








AD-
cscsuc
1758
asUfsg
2023
UACCUC
2288


568965.1
uuCfaU

ucUfgU

UUCAUC




fCfCfa

fCfugg

CAGACA




gacaga

aUfgAf

GACAA




cauL96

agaggs








usa








AD-
csuscu
1759
asUfsu
2024
ACCUCU
2289


568966.1
ucAfuC

guCfuG

UCAUCC




fCfAfg

fUfcug

AGACAG




acagac

gAfuGf

ACAAG




aauL96

aagags








gsu








AD-
uscsuu
1760
asCfsu
2025
CCUCUU
2290


568967.1
caUfcC

ugUfcU

CAUCCA




fAfGfa

fGfucu

GACAGA




cagaca

gGfaUf

CAAGA




aguL96

gaagas








gsg








AD-
csusuc
1761
asUfsc
2026
CUCUUC
2291


568968.1
auCfcA

uuGfuC

AUCCAG




fGfAfc

fUfguc

ACAGAC




agacaa

uGfgAf

AAGAC




gauL96

ugaags








asg








AD-
ususca
1762
asGfsu
2027
UCUUCA
2292


568969.1
ucCfaG

cuUfgU

UCCAGA




fAfCfa

fCfugu

CAGACA




gacaag

cUfgGf

AGACC




acuL96

augaas








gsa








AD-
uscsau
1763
asGfsg
2028
CUUCAU
2293


568970.1
ccAfgA

ucUfuG

CCAGAC




fCfAfg

fUfcug

AGACAA




acaaga

uCfuGf

GACCA




ccuL96

gaugas








asg








AD-
csasuc
1764
asUfsg
2029
UUCAUC
2294


568971.1
caGfaC

guCfuU

CAGACA




fAfGfa

fGfucu

GACAAG




caagac

gUfcUf

ACCAU




cauL96

ggaugs








asa








AD-
asuscc
1765
asAfsu
2030
UCAUCC
2295


568972.1
agAfcA

ggUfcU

AGACAG




fGfAfc

fUfguc

ACAAGA




aagacc

uGfuCf

CCAUC




auuL96

uggaus








gsa








AD-
uscsca
1766
asGfsa
2031
CAUCCA
2296


568973.1
gaCfaG

ugGfuC

GACAGA




fAfCfa

fUfugu

CAAGAC




agacca

cUfgUf

CAUCU




ucuL96

cuggas








usg








AD-
cscsag
1767
asAfsg
2032
AUCCAG
2297


568974.1
acAfgA

auGfgU

ACAGAC




fCfAfa

fCfuug

AAGACC




gaccau

uCfuGf

AUCUA




cuuL96

ucuggs








asu








AD-
csasga
1768
asUfsa
2033
UCCAGA
2298


568975.1
caGfaC

gaUfgG

CAGACA




fAfAfg

fUfcuu

AGACCA




accauc

gUfcUf

UCUAC




uauL96

gucugs








gsa








AD-
gsasca
1769
asUfsg
2034
CAGACA
2299


568977.1
gaCfaA

uaGfaU

GACAAG




fGfAfc

fGfguc

ACCAUC




caucua

uUfgUf

UACAC




cauL96

cugucs








usg








AD-
csasga
1770
asGfsg
2035
GACAGA
2300


568979.1
caAfgA

ugUfaG

CAAGAC




fCfCfa

fAfugg

CAUCUA




ucuaca

uCfuUf

CACCC




ccuL96

gucugs








usc








AD-
asgsac
1771
asGfsg
2036
ACAGAC
2301


1069834.1
aaGfaC

guGfuA

AAGACC




fCfAfu

fGfaug

AUCUAC




cuacac

gUfcUf

ACCCC




ccuL96

ugucus








gsu








AD-
gsasca
1772
asGfsg
2037
CAGACA
2302


1069835.1
agAfcC

ggUfgU

AGACCA




fAfUfc

fAfgau

UCUACA




uacacc

gGfuCf

CCCCU




ccuL96

uugucs








usg








AD-
ascsaa
1773
asAfsg
2038
AGACAA
2303


1069836.1
gaCfcA

ggGfuG

GACCAU




fUfCfu

fUfaga

CUACAC




acaccc

uGfgUf

CCCUG




cuuL96

cuugus








csu








AD-
gsgscc
1774
asGfsc
2039
UGGGCC
2304


569154.1
agUfgG

ucGfgA

AGUGGA




fAfAfg

fUfcuu

AGAUCC




auccga

cCfaCf

GAGCC




gcuL96

uggccs








csa








AD-
gscsca
1775
asGfsg
2040
GGGCCA
2305


569155.1
guGfgA

cuCfgG

GUGGAA




fAfGfa

fAfucu

GAUCCG




uccgag

uCfcAf

AGCCU




ccuL96

cuggcs








csc








AD-
cscsag
1776
asAfsg
2041
GGCCAG
2306


569156.1
ugGfaA

gcUfcG

UGGAAG




fGfAfu

fGfauc

AUCCGA




ccgagc

uUfcCf

GCCUA




cuuL96

acuggs








csc








AD-
csasgu
1777
asUfsa
2042
GCCAGU
2307


569157.1
ggAfaG

ggCfuC

GGAAGA




fAfUfc

fGfgau

UCCGAG




cgagcc

cUfuCf

CCUAC




uauL96

cacugs








gsc








AD-
asgsug
1778
asGfsu
2043
CCAGUG
2308


569158.1
gaAfgA

agGfcU

GAAGAU




fUfCfc

fCfgga

CCGAGC




gagccu

uCfuUf

CUACU




acuL96

ccacus








gsg








AD-
gsusgg
1779
asAfsg
2044
CAGUGG
2309


569159.1
aaGfaU

uaGfgC

AAGAUC




fCfCfg

fUfcgg

CGAGCC




agccua

aUfcUf

UACUA




cuuL96

uccacs








usg








AD-
usgsga
1780
asUfsa
2045
AGUGGA
2310


569160.1
agAfuC

guAfgG

AGAUCC




fCfGfa

fCfucg

GAGCCU




gccuac

gAfuCf

ACUAU




uauL96

uuccas








csu








AD-
gsgsaa
1781
asAfsu
2046
GUGGAA
2311


569161.1
gaUfcC

agUfaG

GAUCCG




fGfAfg

fGfcuc

AGCCUA




ccuacu

gGfaUf

CUAUG




auuL96

cuuccs








asc








AD-
gsasag
1782
asCfsa
2047
UGGAAG
2312


569162.1
auCfcG

uaGfuA

AUCCGA




fAfGfc

fGfgcu

GCCUAC




cuacua

cGfgAf

UAUGA




uguL96

ucuucs








csa








AD-
asasga
1783
asUfsc
2048
GGAAGA
2313


569163.1
ucCfgA

auAfgU

UCCGAG




fGfCfc

fAfggc

CCUACU




uacuau

uCfgGf

AUGAA




gauL96

aucuus








csc








AD-
asuscc
1784
asUfsu
2049
AGAUCC
2314


569166.1
gaGfcC

uuCfaU

GAGCCU




fUfAfc

fAfgua

ACUAUG




uaugaa

gGfcUf

AAAAC




aauL96

cggaus








csu








AD-
usescg
1785
asGfsu
2050
GAUCCG
2315


569167.1
agCfcU

uuUfcA

AGCCUA




fAfCfu

fUfagu

CUAUGA




augaaa

aGfgCf

AAACU




acuL96

ucggas








usc








AD-
cscsga
1786
asAfsg
2051
AUCCGA
2316


569168.1
gcCfuA

uuUfuC

GCCUAC




fCfUfa

fAfuag

UAUGAA




ugaaaa

uAfgGf

AACUC




cuuL96

cucggs








asu








AD-
csgsag
1787
asGfsa
2052
UCCGAG
2317


569169.1
ccUfaC

guUfuU

CCUACU




fUfAfu

fCfaua

AUGAAA




gaaaac

gUfaGf

ACUCA




ucuL96

gcucgs








gsa








AD-
gsasgc
1788
asUfsg
2053
CCGAGC
2318


569170.1
cuAfcU

agUfuU

CUACUA




fAfUfg

fUfcau

UGAAAA




aaaacu

aGfuAf

CUCAC




cauL96

ggcucs








gsg








AD-
asgscc
1789
asGfsu
2054
CGAGCC
2319


569171.1
uaCfuA

gaGfuU

UACUAU




fUfGfa

fUfuca

GAAAAC




aaacuc

uAfgUf

UCACC




acuL96

aggcus








csg








AD-
gscscu
1790
asGfsg
2055
GAGCCU
2320


569172.1
acUfaU

ugAfgU

ACUAUG




fGfAfa

fUfuuc

AAAACU




aacuca

aUfaGf

CACCA




ccuL96

uaggcs








usc








AD-
cscsua
1791
asUfsg
2056
AGCCUA
2321


569173.1
cuAfuG

guGfaG

CUAUGA




fAfAfa

fUfuuu

AAACUC




acucac

cAfuAf

ACCAC




cauL96

guaggs








csu








AD-
csusac
1792
asGfsu
2057
GCCUAC
2322


569174.1
uaUfgA

ggUfgA

UAUGAA




fAfAfa

fGfuuu

AACUCA




cucacc

uCfaUf

CCACA




acuL96

aguags








gsc








AD-
usascu
1793
asUfsg
2058
CCUACU
2323


569175.1
auGfaA

ugGfuG

AUGAAA




fAfAfc

fAfguu

ACUCAC




ucacca

uUfcAf

CACAG




cauL96

uaguas








gsg








AD-
cscsua
1794
asUfsa
2059
AGCCUA
2324


569262.1
caGfaG

guAfgA

CAGAGA




fAfAfa

fAfuuu

AAUUCU




uucuac

cUfcUf

ACUAC




uauL96

guaggs








csu








AD-
csusac
1795
asGfsu
2060
GCCUAC
2325


569263.1
agAfgA

agUfaG

AGAGAA




fAfAfu

fAfauu

AUUCUA




ucuacu

uCfuCf

CUACA




acuL96

uguags








gsc








AD-
usasca
1796
asUfsg
2061
CCUACA
2326


569264.1
gaGfaA

uaGfuA

GAGAAA




fAfUfu

fGfaau

UUCUAC




cuacua

uUfcUf

UACAU




cauL96

cuguas








gsg








AD-
ascsag
1797
asAfsu
2062
CUACAG
2327


569265.1
agAfaA

guAfgU

AGAAAU




fUfUfc

fAfgaa

UCUACU




uacuac

uUfuCf

ACAUC




auuL96

ucugus








asg








AD-
csasga
1798
asGfsa
2063
UACAGA
2328


569266.1
gaAfaU

ugUfaG

GAAAUU




fUfCfu

fUfaga

CUACUA




acuaca

aUfuUf

CAUCU




ucuL96

cucugs








usa








AD-
asgsag
1799
asAfsg
2064
ACAGAG
2329


569267.1
aaAfuU

auGfuA

AAAUUC




fCfUfa

fGfuag

UACUAC




cuacau

aAfuUf

AUCUA




cuuL96

ucucus








gsu








AD-
gsasga
1800
asUfsa
2065
CAGAGA
2330


569268.1
aaUfuC

gaUfgU

AAUUCU




fUfAfc

fAfgua

ACUACA




uacauc

gAfaUf

UCUAU




uauL96

uucucs








usg








AD-
asgsaa
1801
asAfsu
2066
AGAGAA
2331


569269.1
auUfcU

agAfuG

AUUCUA




fAfCfu

fUfagu

CUACAU




acaucu

aGfaAf

CUAUA




auuL96

uuucus








csu








AD-
gsasaa
1802
asUfsa
2067
GAGAAA
2332


569270.1
uuCfuA

uaGfaU

UUCUAC




fCfUfa

fGfuag

UACAUC




caucua

uAfgAf

UAUAA




uauL96

auuucs








usc








AD-
asasau
1803
asUfsu
2068
AGAAAU
2333


569271.1
ucUfaC

auAfgA

UCUACU




fUfAfc

fUfgua

ACAUCU




aucuau

gUfaGf

AUAAC




aauL96

aauuus








csu








AD-
asusuc
1804
asCfsg
2069
AAAUUC
2334


569273.1
uaCfuA

uuAfuA

UACUAC




fCfAfu

fGfaug

AUCUAU




cuauaa

uAfgUf

AACGA




cguL96

agaaus








usu








AD-
ususcu
1805
asUfsc
2070
AAUUCU
2335


569274.1
acUfaC

guUfaU

ACUACA




fAfUfc

fAfgau

UCUAUA




uauaac

gUfaGf

ACGAG




gauL96

uagaas








usu








AD-
uscsua
1806
asCfsu
2071
AUUCUA
2336


569275.1
cuAfcA

cgUfuA

CUACAU




fUfCfu

fUfaga

CUAUAA




auaacg

uGfuAf

CGAGA




aguL96

guagas








asu








AD-
csusac
1807
asUfsc
2072
UUCUAC
2337


569276.1
uaCfaU

ucGfuU

UACAUC




fCfUfa

fAfuag

UAUAAC




uaacga

aUfgUf

GAGAA




gauL96

aguags








asa








AD-
usascu
1808
asUfsu
2073
UCUACU
2338


569277.1
acAfuC

cuCfgU

ACAUCU




fUfAfu

fUfaua

AUAACG




aacgag

gAfuGf

AGAAG




aauL96

uaguas








gsa








AD-
ascsua
1809
asCfsu
2074
CUACUA
2339


569278.1
caUfcU

ucUfcG

CAUCUA




fAfUfa

fUfuau

UAACGA




acgaga

aGfaUf

GAAGG




aguL96

guagus








asg








AD-
csusac
1810
asCfsc
2075
UACUAC
2340


569279.1
auCfuA

uuCfuC

AUCUAU




fUfAfa

fGfuua

AACGAG




cgagaa

uAfgAf

AAGGG




gguL96

uguags








usa








AD-
usasca
1811
asCfsc
2076
ACUACA
2341


569280.1
ucUfaU

cuUfcU

UCUAUA




fAfAfc

fCfguu

ACGAGA




gagaag

aUfaGf

AGGGC




gguL96

auguas








gsu








AD-
ascsau
1812
asGfsc
2077
CUACAU
2342


569281.1
cuAfuA

ccUfuC

CUAUAA




fAfCfg

fUfcgu

CGAGAA




agaagg

uAfuAf

GGGCC




gcuL96

gaugus








asg








AD-
csasuc
1813
asGfsg
2078
UACAUC
2343


569282.1
uaUfaA

ccCfuU

UAUAAC




fCfGfa

fCfucg

GAGAAG




gaaggg

uUfaUf

GGCCU




ccuL96

agaugs








usa








AD-
cscsuc
1814
asGfsu
2079
GACCUC
2344


569506.1
ucCfcU

ggAfuC

UCCCUA




fAfCfc

fUfggu

CCAGAU




agaucc

aGfgGf

CCACU




acuL96

agaggs








usc








AD-
csuscu
1815
asAfsg
2080
ACCUCU
2345


569507.1
ccCfuA

ugGfaU

CCCUAC




fCfCfa

fCfugg

CAGAUC




gaucca

uAfgGf

CACUU




cuuL96

gagags








gsu








AD-
uscsuc
1816
asAfsa
2081
CCUCUC
2346


569508.1
ccUfaC

guGfgA

CCUACC




fCfAfg

fUfcug

AGAUCC




auccac

gUfaGf

ACUUC




uuuL96

ggagas








gsg








AD-
csuscc
1817
asGfsa
2082
CUCUCC
2347


569509.1
cuAfcC

agUfgG

CUACCA




fAfGfa

fAfucu

GAUCCA




uccacu

gGfuAf

CUUCA




ucuL96

gggags








asg








AD-
uscscc
1818
asUfsg
2083
UCUCCC
2348


569510.1
uaCfcA

aaGfuG

UACCAG




fGfAfu

fGfauc

AUCCAC




ccacuu

uGfgUf

UUCAC




cauL96

agggas








gsa








AD-
cscscu
1819
asGfsu
2084
CUCCCU
2349


569511.1
acCfaG

gaAfgU

ACCAGA




fAfUfc

fGfgau

UCCACU




cacuuc

cUfgGf

UCACC




acuL96

uagggs








asg








AD-
cscsua
1820
asGfsg
2085
UCCCUA
2350


569512.1
ccAfgA

ugAfaG

CCAGAU




fUfCfc

fUfgga

CCACUU




acuuca

uCfuGf

CACCA




ccuL96

guaggs








gsa








AD-
csusac
1821
asUfsg
2086
CCCUAC
2351


569513.1
caGfaU

guGfaA

CAGAUC




fCfCfa

fGfugg

CACUUC




cuucac

aUfcUf

ACCAA




cauL96

gguags








gsg








AD-
usascc
1822
asUfsu
2087
CCUACC
2352


569514.1
agAfuC

ggUfgA

AGAUCC




fCfAfc

fAfgug

ACUUCA




uucacc

gAfuCf

CCAAG




aauL96

ugguas








gsg








AD-
ascsca
1823
asCfsu
2088
CUACCA
2353


569515.1
gaUfcC

ugGfuG

GAUCCA




fAfCfu

fAfagu

CUUCAC




ucacca

gGfaUf

CAAGA




aguL96

cuggus








asg








AD-
cscsag
1824
asUfsc
2089
UACCAG
2354


569516.1
auCfcA

uuGfgU

AUCCAC




fCfUfu

fGfaag

UUCACC




caccaa

uGfgAf

AAGAC




gauL96

ucuggs








usa








AD-
csasga
1825
asGfsu
2090
ACCAGA
2355


569517.1
ucCfaC

cuUfgG

UCCACU




fUfUfc

fUfgaa

UCACCA




accaag

gUfgGf

AGACA




acuL96

aucugs








gsu








AD-
asgsau
1826
asUfsg
2091
CCAGAU
2356


569518.1
ccAfcU

ucUfuG

CCACUU




fUfCfa

fGfuga

CACCAA




ccaaga

aGfuGf

GACAC




cauL96

gaucus








gsg








AD-
gsasuc
1827
asGfsu
2092
CAGAUC
2357


569519.1
caCfuU

guCfuU

CACUUC




fCfAfc

fGfgug

ACCAAG




caagac

aAfgUf

ACACC




acuL96

ggaucs








usg








AD-
asuscc
1828
asGfsg
2093
AGAUCC
2358


569520.1
acUfuC

ugUfcU

ACUUCA




fAfCfc

fUfggu

CCAAGA




aagaca

gAfaGf

CACCC




ccuL96

uggaus








csu








AD-
ususug
1829
asAfsc
2094
CCUUUG
2359


569565.1
acCfuC

gaAfcA

ACCUCA




fAfUfg

fCfcau

UGGUGU




guguuc

gAfgGf

UCGUG




guuL96

ucaaas








gsg








AD-
usgsac
1830
asUfsc
2095
UUUGAC
2360


569567.1
cuCfaU

acGfaA

CUCAUG




fGfGfu

fCfacc

GUGUUC




guucgu

aUfgAf

GUGAC




gauL96

ggucas








asa








AD-
asgsgg
1831
asAfsu
2096
CAAGGG
2361


570126.1
cgUfgU

ucAfgC

CGUGUU




fUfCfg

fAfcga

CGUGCU




ugcuga

aCfaCf

GAAUA




auuL96

gcccus








usg








AD-
gsgsgc
1832
asUfsa
2097
AAGGGC
2362


570127.1
guGfuU

uuCfaG

GUGUUC




fCfGfu

fCfacg

GUGCUG




gcugaa

aAfcAf

AAUAA




uauL96

cgcccs








usu








AD-
gsgscg
1833
asUfsu
2098
AGGGCG
2363


570128.1
ugUfuC

auUfcA

UGUUCG




fGfUfg

fGfcac

UGCUGA




cugaau

gAfaCf

AUAAG




aauL96

acgccs








csu








AD-
gscsgu
1834
asCfsu
2099
GGGCGU
2364


570129.1
guUfcG

uaUfuC

GUUCGU




fUfGfc

fAfgca

GCUGAA




ugaaua

cGfaAf

UAAGA




aguL96

cacgcs








csc








AD-
gsusgu
1835
asUfsu
2100
GCGUGU
2365


570131.1
ucGfuG

cuUfaU

UCGUGC




fCfUfg

fUfcag

UGAAUA




aauaag

cAfcGf

AGAAG




aauL96

aacacs








gsc








AD-
uscsgu
1836
asGfsu
2101
GUUCGU
2366


570135.1
gcUfgA

ucUfuC

GCUGAA




fAfUfa

fUfuau

UAAGAA




agaaga

uCfaGf

GAACA




acuL96

cacgas








asc








AD-
csgsug
1837
asUfsg
2102
UUCGUG
2367


570136.1
cuGfaA

uuCfuU

CUGAAU




fUfAfa

fCfuua

AAGAAG




gaagaa

uUfcAf

AACAA




cauL96

gcacgs








asa








AD-
asgsac
1838
asAfsg
2103
GGAGAC
2368


571535.1
uuCfcU

uuGfgC

UUCCUU




fUfGfa

fUfuca

GAAGCC




agccaa

aGfgAf

AACUA




cuuL96

agucus








csc








AD-
gsascu
1839
asUfsa
2104
GAGACU
2369


571536.1
ucCfuU

guUfgG

UCCUUG




fGfAfa

fCfuuc

AAGCCA




gccaac

aAfgGf

ACUAC




uauL96

aagucs








usc








AD-
ascsuu
1840
asGfsu
2105
AGACUU
2370


571537.1
ccUfuG

agUfuG

CCUUGA




fAfAfg

fGfcuu

AGCCAA




ccaacu

cAfaGf

CUACA




acuL96

gaagus








csu








AD-
csusuc
1841
asUfsg
2106
GACUUC
2371


571538.1
cuUfgA

uaGfuU

CUUGAA




fAfGfc

fGfgcu

GCCAAC




caacua

uCfaAf

UACAU




cauL96

ggaags








usc








AD-
uscscu
1842
asCfsa
2107
CUUCCU
2372


571540.1
ugAfaG

ugUfaG

UGAAGC




fCfCfa

fUfugg

CAACUA




acuaca

cUfuCf

CAUGA




uguL96

aaggas








asg








AD-
cscsuu
1843
asUfsc
2108
UUCCUU
2373


571541.1
gaAfgC

auGfuA

GAAGCC




fCfAfa

fGfuug

AACUAC




cuacau

gCfuUf

AUGAA




gauL96

caaggs








asa








AD-
csusug
1844
asUfsu
2109
UCCUUG
2374


571542.1
aaGfcC

caUfgU

AAGCCA




fAfAfc

fAfguu

ACUACA




uacaug

gGfcUf

UGAAC




aauL96

ucaags








gsa








AD-
ususga
1845
asGfsu
2110
CCUUGA
2375


571543.1
agCfcA

ucAfuG

AGCCAA




fAfCfu

fUfagu

CUACAU




acauga

uGfgCf

GAACC




acuL96

uucaas








gsg








AD-
usgsaa
1846
asGfsg
2111
CUUGAA
2376


571544.1
gcCfaA

uuCfaU

GCCAAC




fCfUfa

fGfuag

UACAUG




caugaa

uUfgGf

AACCU




ccuL96

cuucas








asg








AD-
gsasag
1847
asAfsg
2112
UUGAAG
2377


571545.1
ccAfaC

guUfcA

CCAACU




fUfAfc

fUfgua

ACAUGA




augaac

gUfuGf

ACCUA




cuuL96

gcuucs








asa








AD-
asasgc
1848
asUfsa
2113
UGAAGC
2378


571546.1
caAfcU

ggUfuC

CAACUA




fAfCfa

fAfugu

CAUGAA




ugaacc

aGfuUf

CCUAC




uauL96

ggcuus








csa








AD-
asgscc
1849
asGfsu
2114
GAAGCC
2379


571547.1
aaCfuA

agGfuU

AACUAC




fCfAfu

fCfaug

AUGAAC




gaaccu

uAfgUf

CUACA




acuL96

uggcus








usc








AD-
gscsca
1850
asUfsg
2115
AAGCCA
2380


571548.1
acUfaC

uaGfgU

ACUACA




fAfUfg

fUfcau

UGAACC




aaccua

gUfaGf

UACAG




cauL96

uuggcs








usu








AD-
cscsaa
1851
asCfsu
2116
AGCCAA
2381


571549.1
cuAfcA

guAfgG

CUACAU




fUfGfa

fUfuca

GAACCU




accuac

uGfuAf

ACAGA




aguL96

guuggs








csu








AD-
csasac
1852
asUfsc
2117
GCCAAC
2382


571550.1
uaCfaU

ugUfaG

UACAUG




fGfAfa

fGfuuc

AACCUA




ccuaca

aUfgUf

CAGAG




gauL96

aguugs








gsc








AD-
asascu
1853
asCfsu
2118
CCAACU
2383


571551.1
acAfuG

cuGfuA

ACAUGA




fAfAfc

fGfguu

ACCUAC




cuacag

cAfuGf

AGAGA




aguL96

uaguus








gsg








AD-
ascsua
1854
asUfsc
2119
CAACUA
2384


571552.1
caUfgA

ucUfgU

CAUGAA




fAfCfc

fAfggu

CCUACA




uacaga

uCfaUf

GAGAU




gauL96

guagus








usg








AD-
csusac
1855
asAfsu
2120
AACUAC
2385


571553.1
auGfaA

cuCfuG

AUGAAC




fCfCfu

fUfagg

CUACAG




acagag

uUfcAf

AGAUC




auuL96

uguags








usu








AD-
usasca
1856
asGfsa
2121
ACUACA
2386


571554.1
ugAfaC

ucUfcU

UGAACC




fCfUfa

fGfuag

UACAGA




cagaga

gUfuCf

GAUCC




ucuL96

auguas








gsu








AD-
ascsau
1857
asGfsg
2122
CUACAU
2387


571555.1
gaAfcC

auCfuC

GAACCU




fUfAfc

fUfgua

ACAGAG




agagau

gGfuUf

AUCCU




ccuL96

caugus








asg








AD-
csasug
1858
asAfsg
2123
UACAUG
2388


571556.1
aaCfcU

gaUfcU

AACCUA




fAfCfa

fCfugu

CAGAGA




gagauc

aGfgUf

UCCUA




cuuL96

ucaugs








usa








AD-
asusga
1859
asUfsa
2124
ACAUGA
2389


571557.1
acCfuA

ggAfuC

ACCUAC




fCfAfg

fUfcug

AGAGAU




agaucc

uAfgGf

CCUAC




uauL96

uucaus








gsu








AD-
usgsaa
1860
asGfsu
2125
CAUGAA
2390


571558.1
ccUfaC

agGfaU

CCUACA




fAfGfa

fCfucu

GAGAUC




gauccu

gUfaGf

CUACA




acuL96

guucas








usg








AD-
gsasac
1861
asUfsg
2126
AUGAAC
2391


571559.1
cuAfcA

uaGfgA

CUACAG




fGfAfg

fUfcuc

AGAUCC




auccua

uGfuAf

UACAC




cauL96

gguucs








asu








AD-
asascc
1862
asGfsu
2127
UGAACC
2392


571560.1
uaCfaG

guAfgG

UACAGA




fAfGfa

fAfucu

GAUCCU




uccuac

cUfgUf

ACACU




acuL96

agguus








csa








AD-
gsgscc
1863
asCfsu
2128
UUGGCC
2393


571711.1
cuAfcU

uuUfaG

CUACUG




fGfCfa

fCfugc

CAGCUA




gcuaaa

aGfuAf

AAAGA




aguL96

gggccs








asa








AD-
gscscc
1864
asUfsc
2129
UGGCCC
2394


571712.1
uaCfuG

uuUfuA

UACUGC




fCfAfg

fGfcug

AGCUAA




cuaaaa

cAfgUf

AAGAC




gauL96

agggcs








csa








AD-
cscscu
1865
asGfsu
2130
GGCCCU
2395


571713.1
acUfgC

cuUful

ACUGCA




fAfGfc

lfAfgc

GCUAAA




uaaaag

ugCfaG

AGACU




acuL96

fuaggg








scsc








AD-
cscsua
1866
asAfsg
2131
GCCCUA
2396


571714.1
cuGfcA

ucUful

CUGCAG




fGfCfu

lfUfag

CUAAAA




aaaaga

cuGfcA

GACUU




cuuL96

fguagg








sgsc








AD-
usascu
1867
asAfsa
2132
CCUACU
2397


571716.1
gcAfgC

agUfcU

GCAGCU




fUfAfa

fUfuua

AAAAGA




aagacu

gCfuGf

CUUUG




uuuL96

caguas








gsg








AD-
ascsug
1868
asCfsa
2133
CUACUG
2398


571717.1
caGfcU

aaGfuC

CAGCUA




fAfAfa

fUfuuu

AAAGAC




agacuu

aGfcUf

UUUGA




uguL96

gcagus








asg








AD-
csusgc
1869
asUfsc
2134
UACUGC
2399


571718.1
agCfuA

aaAfgU

AGCUAA




fAfAfa

fCfuuu

AAGACU




gacuuu

uAfgCf

UUGAC




gauL96

ugcags








usa








AD-
usgsca
1870
asGfsu
2135
ACUGCA
2400


571719.2
gcUfaA

caAfaG

GCUAAA




fAfAfg

fUfcuu

AGACUU




acuuug

uUfaGf

UGACU




acuL96

cugcas








gsu








AD-
gscsag
1871
asAfsg
2136
CUGCAG
2401


571720.1
cuAfaA

ucAfaA

CUAAAA




fAfGfa

fGfucu

GACUUU




cuuuga

uUfuAf

GACUU




cuuL96

gcugcs








asg








AD-
csasgc
1872
asAfsa
2137
UGCAGC
2402


571721.1
uaAfaA

guCfaA

UAAAAG




fGfAfc

fAfguc

ACUUUG




uuugac

uUfull

ACUUU




uuuL96

fagcug








scsa








AD-
asgscu
1873
asAfsa
2138
GCAGCU
2403


571722.1
aaAfaG

agUfcA

AAAAGA




fAfCfu

fAfagu

CUUUGA




uugacu

cUfuUf

CUUUG




uuuL96

uagcus








gsc








AD-
gscsua
1874
asCfsa
2139
CAGCUA
2404


571723.1
aaAfgA

aaGfuC

AAAGAC




fCfUfu

fAfaag

UUUGAC




ugacuu

uCfuUf

UUUGU




uguL96

uuagcs








usg








AD-
gsusgc
1875
asCfsa
2140
UUGUGC
2405


571742.1
cuCfcC

acGfcA

CUCCCG




fGfUfc

fCfgac

UCGUGC




gugcgu

gGfgAf

GUUGG




uguL96

ggcacs








asa








AD-
usgscc
1876
asCfsc
2141
UGUGCC
2406


571743.1
ucCfcG

aaCfgC

UCCCGU




fUfCfg

fAfcga

CGUGCG




ugcguu

cGfgGf

UUGGC




gguL96

aggcas








csa








AD-
gscscu
1877
asGfsc
2142
GUGCCU
2407


571744.1
ccCfgU

caAfcG

CCCGUC




fCfGfu

fCfacg

GUGCGU




gcguug

aCfgGf

UGGCU




gcuL96

gaggcs








asc








AD-
cscsuc
1878
asAfsg
2143
UGCCUC
2408


571745.1
ccGfuC

ccAfaC

CCGUCG




fGfUfg

fGfcac

UGCGUU




cguugg

gAfcGf

GGCUC




cuuL96

ggaggs








csa








AD-
csuscc
1879
asGfsa
2144
GCCUCC
2409


571746.1
cgUfcG

gcCfaA

CGUCGU




fUfGfc

fCfgca

GCGUUG




guuggc

cGfaCf

GCUCA




ucuL96

gggags








gsc








AD-
uscscc
1880
asUfsg
2145
CCUCCC
2410


571747.1
guCfgU

agCfcA

GUCGUG




fGfCfg

fAfcgc

CGUUGG




uuggcu

aCfgAf

CUCAA




cauL96

cgggas








gsg








AD-
csescg
1881
asUfsu
2146
CUCCCG
2411


571748.1
ucGfuG

gaGfcC

UCGUGC




fCfGfu

fAfacg

GUUGGC




uggcuc

cAfcGf

UCAAU




aauL96

acgggs








asg








AD-
cscsgu
1882
asAfsu
2147
UCCCGU
2412


571749.1
cgUfgC

ugAfgC

CGUGCG




fGfUfu

fCfaac

UUGGCU




ggcuca

gCfaCf

CAAUG




auuL96

gacggs








gsa








AD-
csgsuc
1883
asCfsa
2148
CCCGUC
2413


571750.1
guGfcG

uuGfaG

GUGCGU




fUfUfg

fCfcaa

UGGCUC




gcucaa

cGfcAf

AAUGA




uguL96

cgacgs








gsg








AD-
gsuscg
1884
asUfsc
2149
CCGUCG
2414


571751.1
ugCfgU

auUfgA

UGCGUU




fUfGfg

fGfcca

GGCUCA




cucaau

aCfgCf

AUGAA




gauL96

acgacs








gsg








AD-
csgsug
1885
asGfsu
2150
GUCGUG
2415


571753.2
egUfuG

ucAfuU

CGUUGG




fGfCfu

fGfagc

CUCAAU




caauga

cAfaCf

GAACA




acuL96

gcacgs








asc








AD-
usgscg
1886
asCfsu
2151
CGUGCG
2416


571755.1
uuGfgC

guUfcA

UUGGCU




fUfCfa

fUfuga

CAAUGA




augaac

gCfcAf

ACAGA




aguL96

acgcas








csg








AD-
gscsgu
1887
asUfsc
2152
GUGCGU
2417


571756.1
ugGfcU

ugUfuC

UGGCUC




fCfAfa

fAfuug

AAUGAA




ugaaca

aGfcCf

CAGAG




gauL96

aacgcs








asc








AD-
csgsuu
1888
asCfsu
2153
UGCGUU
2418


571757.1
ggCfuC

cuGfuU

GGCUCA




fAfAfu

fCfauu

AUGAAC




gaacag

gAfgCf

AGAGA




aguL96

caacgs








csa








AD-
gsusug
1889
asUfsc
2154
GCGUUG
2419


571758.1
gcUfcA

ucUfgU

GCUCAA




fAfUfg

fUfcau

UGAACA




aacaga

uGfaGf

GAGAU




gauL96

ccaacs








gsc








AD-
ususgg
1890
asAfsu
2155
CGUUGG
2420


571759.1
cuCfaA

cuCfuG

CUCAAU




fUfGfa

fUfuca

GAACAG




acagag

uUfgAf

AGAUA




auuL96

gccaas








csg








AD-
usgsgc
1891
asUfsa
2156
GUUGGC
2421


571760.1
ucAfaU

ucUfcU

UCAAUG




fGfAfa

fGfuuc

AACAGA




cagaga

aUfuGf

GAUAC




uauL96

agccas








asc








AD-
gsgscu
1892
asGfsu
2157
UUGGCU
2422


571761.1
caAfuG

auCfuC

CAAUGA




fAfAfc

fUfguu

ACAGAG




agagau

cAfuUf

AUACU




acuL96

gagccs








asa








AD-
gscsuc
1893
asAfsg
2158
UGGCUC
2423


571762.1
aaUfgA

uaUfcU

AAUGAA




fAfCfa

fCfugu

CAGAGA




gagaua

uCfaUf

UACUA




cuuL96

ugagcs








csa








AD-
csusca
1894
asUfsa
2159
GGCUCA
2424


571763.1
auGfaA

guAfuC

AUGAAC




fCfAfg

fUfcug

AGAGAU




agauac

uUfcAf

ACUAC




uauL96

uugags








csc








AD-
uscsaa
1895
asGfsu
2160
GCUCAA
2425


571764.1
ugAfaC

agUfaU

UGAACA




fAfGfa

fCfucu

GAGAUA




gauacu

gUfuCf

CUACG




acuL96

auugas








gsc








AD-
csasau
1896
asCfsg
2161
CUCAAU
2426


571765.2
gaAfcA

uaGfuA

GAACAG




fGfAfg

fUfcuc

AGAUAC




auacua

uGfuUf

UACGG




cguL96

cauugs








asg








AD-
asasug
1897
asCfsc
2162
UCAAUG
2427


571766.2
aaCfaG

guAfgU

AACAGA




fAfGfa

fAfucu

GAUACU




uacuac

cUfgUf

ACGGU




gguL96

ucauus








gsa








AD-
asusga
1898
asAfsc
2163
CAAUGA
2428


571767.2
acAfgA

cgUfaG

ACAGAG




fGfAfu

fUfauc

AUACUA




acuacg

uCfuGf

CGGUG




guuL96

uucaus








usg








AD-
gscsag
1899
asUfsa
2164
GAGCAG
2429


572383.1
ucAfaG

ggCfgU

UCAAGG




fGfUfc

fAfgac

UCUACG




uacgcc

cUfuGf

CCUAU




uauL96

acugcs








usc








AD-
csasgu
1900
asAfsu
2165
AGCAGU
2430


572384.1
caAfgG

agGfcG

CAAGGU




fUfCfu

fUfaga

CUACGC




acgccu

cCfuUf

CUAUU




auuL96

gacugs








csu








AD-
asgsuc
1901
asAfsa
2166
GCAGUC
2431


572385.1
aaGfgU

uaGfgC

AAGGUC




fCfUfa

fGfuag

UACGCC




cgccua

aCfcUf

UAUUA




uuuL96

ugacus








gsc








AD-
gsusca
1902
asUfsa
2167
CAGUCA
2432


572386.1
agGfuC

auAfgG

AGGUCU




fUfAfc

fCfgua

ACGCCU




gccuau

gAfcCf

AUUAC




uauL96

uugacs








usg








AD-
uscsaa
1903
asGfsu
2168
AGUCAA
2433


572387.4
ggUfcU

aaUfaG

GGUCUA




fAfCfg

fGfcgu

CGCCUA




ccuauu

aGfaCf

UUACA




acuL96

cuugas








csu








AD-
gsgsuc
1904
asGfsg
2169
AAGGUC
2434


572391.1
uaCfgC

uuGfuA

UACGCC




fCfUfa

fAfuag

UAUUAC




uuacaa

gCfgUf

AACCU




ccuL96

agaccs








usu








AD-
gsuscu
1905
asAfsg
2170
AGGUCU
2435


572392.1
acGfcC

guUfgU

ACGCCU




fUfAfu

fAfaua

AUUACA




uacaac

gGfcGf

ACCUG




cuuL96

uagacs








csu








AD-
uscsua
1906
asCfsa
2171
GGUCUA
2436


572393.2
cgCfcU

ggUfuG

CGCCUA




fAfUfu

fUfaau

UUACAA




acaacc

aGfgCf

CCUGG




uguL96

guagas








csc








AD-
csusac
1907
asCfsc
2172
GUCUAC
2437


572394.1
gcCfuA

agGfuU

GCCUAU




fUfUfa

fGfuaa

UACAAC




caaccu

uAfgGf

CUGGA




gguL96

cguags








asc








AD-
usascg
1908
asUfsc
2173
UCUACG
2438


572395.1
ccUfaU

caGfgU

CCUAUU




fUfAfc

fUfgua

ACAACC




aaccug

aUfaGf

UGGAG




gauL96

gcguas








gsa








AD-
ascsgc
1909
asCfsu
2174
CUACGC
2439


572396.1
cuAfuU

ccAfgG

CUAUUA




fAfCfa

fUfugu

CAACCU




accugg

aAfuAf

GGAGG




aguL96

ggcgus








asg








AD-
csgscc
1910
asCfsc
2175
UACGCC
2440


572397.1
uaUfuA

ucCfaG

UAUUAC




fCfAfa

fGfuug

AACCUG




ccugga

uAfaUf

GAGGA




gguL96

aggcgs








usa








AD-
gscsug
1911
asAfsu
2176
GUGCUG
2441


572495.1
agGfaG

gaAfgC

AGGAGA




fAfAfu

fAfauu

AUUGCU




ugcuuc

cUfcCf

UCAUA




auuL96

ucagcs








asc








AD-
gscsca
1912
asAfsc
2177
GAGCCA
2442


572569.1
ggAfgU

acAfuA

GGAGUG




fGfGfa

fGfucc

GACUAU




cuaugu

aCfuCf

GUGUA




guuL96

cuggcs








usc








AD-
cscsag
1913
asUfsa
2178
AGCCAG
2443


572570.1
gaGfuG

caCfaU

GAGUGG




fGfAfc

fAfguc

ACUAUG




uaugug

cAfcUf

UGUAC




uauL96

ccuggs








csu








AD-
csasgg
1914
asGfsu
2179
GCCAGG
2444


572571.1
agUfgG

acAfcA

AGUGGA




fAfCfu

fUfagu

CUAUGU




augugu

cCfaCf

GUACA




acuL96

uccugs








gsc








AD-
asgsga
1915
asUfsg
2180
CCAGGA
2445


572572.1
guGfgA

uaCfaC

GUGGAC




fCfUfa

fAfuag

UAUGUG




ugugua

uCfcAf

UACAA




cauL96

cuccus








gsg








AD-
gsgsag
1916
asUfsu
2181
CAGGAG
2446


572573.1
ugGfaC

guAfcA

UGGACU




fUfAfu

fCfaua

AUGUGU




guguac

gUfcCf

ACAAG




aauL96

acuccs








usg








AD-
gsasgu
1917
asCfsu
2182
AGGAGU
2447


572574.1
ggAfcU

ugUfaC

GGACUA




fAfUfg

fAfcau

UGUGUA




uguaca

aGfuCf

CAAGA




aguL96

cacucs








csu








AD-
asgsug
1918
asUfsc
2183
GGAGUG
2448


572575.1
gaCfuA

uuGfuA

GACUAU




fUfGfu

fCfaca

GUGUAC




guacaa

uAfgUf

AAGAC




gauL96

ccacus








csc








AD-
gsusgg
1919
asGfsu
2184
GAGUGG
2449


572576.1
acUfaU

cuUfgU

ACUAUG




fGfUfg

fAfcac

UGUACA




uacaag

aUfaGf

AGACC




acuL96

uccacs








usc








AD-
usgsga
1920
asGfsg
2185
AGUGGA
2450


572577.1
cuAfuG

ucUfuG

CUAUGU




fUfGfu

fUfaca

GUACAA




acaaga

cAfuAf

GACCC




ccuL96

guccas








csu








AD-
ascsua
1921
asUfsc
2186
GGACUA
2451


572580.1
ugUfgU

ggGfuC

UGUGUA




fAfCfa

fUfugu

CAAGAC




agaccc

aCfaCf

CCGAC




gauL96

auagus








esc








AD-
csusau
1922
asGfsu
2187
GACUAU
2452


572581.1
guGfuA

cgGfgU

GUGUAC




fCfAfa

fCfuug

AAGACC




gacccg

uAfcAf

CGACU




acuL96

cauags








usc
















TABLE 22







Unmodified Sense and Antisense Strand Sequences of Complement Component C3 dsRNA Agents















SEQ
Range in

SEQ
Range in



Sense
ID
NM_
Antisense 
ID
NM_


Duplex Name
Sequence 5’ to 3’
NO:
000064.3
Sequence 5’ to 3’
NO:
000064.3





AD-564723.1
AGAGCGGGUACCUCUUCAUCU
2453
 470-490
AGAUGAAGAGGUACCCGCUCUGC
2714
 468-490





AD-564724.1
GAGCGGGUACCUCUUCAUCCU
2454
 471-491
AGGAUGAAGAGGUACCCGCUCUG
2715
 469-491





AD-1069838.1
AGCGGGUACCUCUUCAUCCAU
2455
 472-492
AUGGAUGAAGAGGUACCCGCUCU
2716
 470-492





AD-564726.1
GCGGGUACCUCUUCAUCCAGU
2456
 473-493
ACUGGATGAAGAGGUACCCGCUC
2717
 471-493





AD-564727.3
CGGGUACCUCUUCAUCCAGAU
2457
 474-494
AUCUGGAUGAAGAGGUACCCGCU
2718
 472-494





AD-1069839.1
GGGUACCUCUUCAUCCAGACU
2458
 475-495
AGUCUGGAUGAAGAGGUACCCGC
2719
 473-495





AD-1069840.1
GGUACCUCUUCAUCCAGACAU
2459
 476-496
AUGUCUGGAUGAAGAGGUACCCG
2720
 474-496





AD-564730.3
GUACCUCUUCAUCCAGACAGU
2460
 477-497
ACUGUCTGGAUGAAGAGGUACCC
2721
 475-497





AD-1069841.1
UACCUCUUCAUCCAGACAGAU
2461
 478-498
AUCUGUCUGGAUGAAGAGGUACC
2722
 476-498





AD-564732.1
ACCUCUUCAUCCAGACAGACU
2462
 479-499
AGUCUGTCUGGAUGAAGAGGUAC
2723
 477-499





AD-1069842.1
CCUCUUCAUCCAGACAGACAU
2463
 480-500
AUGUCUGUCUGGAUGAAGAGGUA
2724
 478-500





AD-564734.1
CUCUUCAUCCAGACAGACAAU
2464
 481-501
AUUGUCTGUCUGGAUGAAGAGGU
2725
 479-501





AD-1069843.1
UCUUCAUCCAGACAGACAAGU
2465
 482-502
ACUUGUCUGUCUGGAUGAAGAGG
2726
 480-502





AD-564736.1
CUUCAUCCAGACAGACAAGAU
2466
 483-503
AUCUUGTCUGUCUGGAUGAAGAG
2727
 481-503





AD-1069844.1
UUCAUCCAGACAGACAAGACU
2467
 484-504
AGUCUUGUCUGUCUGGAUGAAGA
2728
 482-504





AD-564738.1
UCAUCCAGACAGACAAGACCU
2468
 485-505
AGGUCUTGUCUGUCUGGAUGAAG
2729
 483-505





AD-564739.2
CAUCCAGACAGACAAGACCAU
2469
 486-506
AUGGUCTUGUCUGUCUGGAUGAA
2730
 484-506





AD-1069845.1
AUCCAGACAGACAAGACCAUU
2470
 487-507
AAUGGUCUUGUCUGUCUGGAUGA
2731
 485-507





AD-564741.1
UCCAGACAGACAAGACCAUCU
2471
 488-508
AGAUGGTCUUGUCUGUCUGGAUG
2732
 486-508





AD-1069846.1
CCAGACAGACAAGACCAUCUU
2472
 489-509
AAGAUGGUCUUGUCUGUCUGGAU
2733
 487-509





AD-1069847.1
CAGACAGACAAGACCAUCUAU
2473
 490-510
AUAGAUGGUCUUGUCUGUCUGGA
2734
 488-510





AD-564745.3
GACAGACAAGACCAUCUACAU
2474
 492-512
AUGUAGAUGGUCUUGUCUGUCUG
2735
 490-512





AD-564747.1
CAGACAAGACCAUCUACACCU
2475
 494-514
AGGUGUAGAUGGUCUUGUCUGUC
2736
 492-514





AD-1069850.1
ACAAGACCAUCUACACCCCUU
2476
 497-517
AAGGGGTGUAGAUGGUCUUGUCU
2737
 495-517





AD-1069851.1
GGCCAGUGGAAGAUCCGAGCU
2477
 697-717
AGCUCGGAUCUUCCACUGGCCCA
2738
 695-717





AD-1069852.1
GCCAGUGGAAGAUCCGAGCCU
2478
 698-718
AGGCUCGGAUCUUCCACUGGCCC
2739
 696-718





AD-1069853.1
CCAGUGGAAGAUCCGAGCCUU
2479
 699-719
AAGGCUCGGAUCUUCCACUGGCC
2740
 697-719





AD-564925.1
CAGUGGAAGAUCCGAGCCUAU
2480
 700-720
AUAGGCTCGGAUCUUCCACUGGC
2741
 698-720





AD-1069854.1
AGUGGAAGAUCCGAGCCUACU
2481
 701-721
AGUAGGCUCGGAUCUUCCACUGG
2742
 699-721





AD-1069855.1
GUGGAAGAUCCGAGCCUACUU
2482
 702-722
AAGUAGGCUCGGAUCUUCCACUG
2743
 700-722





AD-1069856.1
UGGAAGAUCCGAGCCUACUAU
2483
 703-723
AUAGUAGGCUCGGAUCUUCCACU
2744
 701-723





AD-564929.1
GGAAGAUCCGAGCCUACUAUU
2484
 704-724
AAUAGUAGGCUCGGAUCUUCCAC
2745
 702-724





AD-564930.1
GAAGAUCCGAGCCUACUAUGU
2485
 705-725
ACAUAGTAGGCUCGGAUCUUCCA
2746
 703-725





AD-1069857.1
AAGAUCCGAGCCUACUAUGAU
2486
 706-726
AUCAUAGUAGGCUCGGAUCUUCC
2747
 704-726





AD-564934.1
AUCCGAGCCUACUAUGAAAAU
2487
 709-729
AUUUUCAUAGUAGGCUCGGAUCU
2748
 707-729





AD-1069858.1
UCCGAGCCUACUAUGAAAACU
2488
 710-730
AGUUUUCAUAGUAGGCUCGGAUC
2749
 708-730





AD-564936.1
CCGAGCCUACUAUGAAAACUU
2489
 711-731
AAGUUUTCAUAGUAGGCUCGGAU
2750
 709-731





AD-564937.1
CGAGCCUACUAUGAAAACUCU
2490
 712-732
AGAGUUTUCAUAGUAGGCUCGGA
2751
 710-732





AD-564938.1
GAGCCUACUAUGAAAACUCAU
2491
 713-733
AUGAGUTUUCAUAGUAGGCUCGG
2752
 711-733





AD-1069859.1
GCCUACUAUGAAAACUCACCU
2492
 715-735
AGGUGAGUUUUCAUAGUAGGCUC
2753
 713-735





AD-564941.1
CCUACUAUGAAAACUCACCAU
2493
 716-736
AUGGUGAGUUUUCAUAGUAGGCU
2754
 714-736





AD-1069860.1
CUACUAUGAAAACUCACCACU
2494
 717-737
AGUGGUGAGUUUUCAUAGUAGGC
2755
 715-737





AD-564943.1
UACUAUGAAAACUCACCACAU
2495
 718-738
AUGUGGTGAGUUUUCAUAGUAGG
2756
 716-738





AD-1069861.1
CCUACAGAGAAAUUCUACUAU
2496
 805-825
AUAGUAGAAUUUCUCUGUAGGCU
2757
 803-825





AD-565031.1
CUACAGAGAAAUUCUACUACU
2497
 806-826
AGUAGUAGAAUUUCUCUGUAGGC
2758
 804-826





AD-565032.1
UACAGAGAAAUUCUACUACAU
2498
 807-827
AUGUAGTAGAAUUUCUCUGUAGG
2759
 805-827





AD-1069862.1
ACAGAGAAAUUCUACUACAUU
2499
 808-828
AAUGUAGUAGAAUUUCUCUGUAG
2760
 806-828





AD-565034.1
CAGAGAAAUUCUACUACAUCU
2500
 809-829
AGAUGUAGUAGAAUUUCUCUGUA
2761
 807-829





AD-565035.1
AGAGAAAUUCUACUACAUCUU
2501
 810-830
AAGAUGTAGUAGAAUUUCUCUGU
2762
 808-830





AD-1069863.1
GAGAAAUUCUACUACAUCUAU
2502
 811-831
AUAGAUGUAGUAGAAUUUCUCUG
2763
 809-831





AD-565037.1
AGAAAUUCUACUACAUCUAUU
2503
 812-832
AAUAGATGUAGUAGAAUUUCUCU
2764
 810-832





AD-565038.1
GAAAUUCUACUACAUCUAUAU
2504
 813-833
AUAUAGAUGUAGUAGAAUUUCUC
2765
 811-833





AD-1069864.1
AAAUUCUACUACAUCUAUAAU
2505
 814-834
AUUAUAGAUGUAGUAGAAUUUCU
2766
 812-834





AD-565041.1
AUUCUACUACAUCUAUAACGU
2506
 816-836
ACGUUATAGAUGUAGUAGAAUUU
2767
 814-836





AD-565042.1
UUCUACUACAUCUAUAACGAU
2507
 817-837
AUCGUUAUAGAUGUAGUAGAAUU
2768
 815-837





AD-565043.1
UCUACUACAUCUAUAACGAGU
2508
 818-838
ACUCGUTAUAGAUGUAGUAGAAU
2769
 816-838





AD-565044.1
CUACUACAUCUAUAACGAGAU
2509
 819-839
AUCUCGTUAUAGAUGUAGUAGAA
2770
 817-839





AD-1069865.1
UACUACAUCUAUAACGAGAAU
2510
 820-840
AUUCUCGUUAUAGAUGUAGUAGA
2771
 818-840





AD-1069866.1
ACUACAUCUAUAACGAGAAGU
2511
 821-841
ACUUCUCGUUAUAGAUGUAGUAG
2772
 819-841





AD-565047.1
CUACAUCUAUAACGAGAAGGU
2512
 822-842
ACCUUCTCGUUAUAGAUGUAGUA
2773
 820-842





AD-1069867.1
UACAUCUAUAACGAGAAGGGU
2513
 823-843
ACCCUUCUCGUUAUAGAUGUAGU
2774
 821-843





AD-565049.1
ACAUCUAUAACGAGAAGGGCU
2514
 824-844
AGCCCUTCUCGUUAUAGAUGUAG
2775
 822-844





AD-565050.1
CAUCUAUAACGAGAAGGGCCU
2515
 825-845
AGGCCCTUCUCGUUAUAGAUGUA
2776
 823-845





AD-565274.1
CCUCUCCCUACCAGAUCCACU
2516
1142-1162
AGUGGATCUGGUAGGGAGAGGUC
2777
1140-1162





AD-565275.1
CUCUCCCUACCAGAUCCACUU
2517
1143-1163
AAGUGGAUCUGGUAGGGAGAGGU
2778
1141-1163





AD-1069868.1
UCUCCCUACCAGAUCCACUUU
2518
1144-1164
AAAGUGGAUCUGGUAGGGAGAGG
2779
1142-1164





AD-1069869.1
CUCCCUACCAGAUCCACUUCU
2519
1145-1165
AGAAGUGGAUCUGGUAGGGAGAG
2780
1143-1165





AD-565278.2
UCCCUACCAGAUCCACUUCAU
2520
1146-1166
AUGAAGTGGAUCUGGUAGGGAGA
2781
1144-1166





AD-1069870.1
CCCUACCAGAUCCACUUCACU
2521
1147-1167
AGUGAAGUGGAUCUGGUAGGGAG
2782
1145-1167





AD-565280.1
CCUACCAGAUCCACUUCACCU
2522
1148-1168
AGGUGAAGUGGAUCUGGUAGGGA
2783
1146-1168





AD-565281.3
CUACCAGAUCCACUUCACCAU
2523
1149-1169
AUGGUGAAGUGGAUCUGGUAGGG
2784
1147-1169





AD-1069871.1
UACCAGAUCCACUUCACCAAU
2524
1150-1170
AUUGGUGAAGUGGAUCUGGUAGG
2785
1148-1170





AD-565283.1
ACCAGAUCCACUUCACCAAGU
2525
1151-1171
ACUUGGTGAAGUGGAUCUGGUAG
2786
1149-1171





AD-1069872.1
CCAGAUCCACUUCACCAAGAU
2526
1152-1172
AUCUUGGUGAAGUGGAUCUGGUA
2787
1150-1172





AD-1069873.1
CAGAUCCACUUCACCAAGACU
2527
1153-1173
AGUCUUGGUGAAGUGGAUCUGGU
2788
1151-1173





AD-565286.1
AGAUCCACUUCACCAAGACAU
2528
1154-1174
AUGUCUTGGUGAAGUGGAUCUGG
2789
1152-1174





AD-565287.1
GAUCCACUUCACCAAGACACU
2529
1155-1175
AGUGUCTUGGUGAAGUGGAUCUG
2790
1153-1175





AD-1069874.1
AUCCACUUCACCAAGACACCU
2530
1156-1176
AGGUGUCUUGGUGAAGUGGAUCU
2791
1154-1176





AD-1069875.1
UUUGACCUCAUGGUGUUCGUU
2531
1201-1221
AACGAACACCAUGAGGUCAAAGG
2792
1199-1221





AD-565335.1
UGACCUCAUGGUGUUCGUGAU
2532
1203-1223
AUCACGAACACCAUGAGGUCAAA
2793
1201-1223





AD-1069876.1
AGGGCGUGUUCGUGCUGAAUU
2533
1892-1912
AAUUCAGCACGAACACGCCCUUG
2794
1890-1912





AD-565895.1
GGGCGUGUUCGUGCUGAAUAU
2534
1893-1913
AUAUUCAGCACGAACACGCCCUU
2795
1891-1913





AD-1069877.1
GGCGUGUUCGUGCUGAAUAAU
2535
1894-1914
AUUAUUCAGCACGAACACGCCCU
2796
1892-1914





AD-565897.1
GCGUGUUCGUGCUGAAUAAGU
2536
1895-1915
ACUUAUTCAGCACGAACACGCCC
2797
1893-1915





AD-565899.1
GUGUUCGUGCUGAAUAAGAAU
2537
1897-1917
AUUCUUAUUCAGCACGAACACGC
2798
1895-1917





AD-565903.1
UCGUGCUGAAUAAGAAGAACU
2538
1901-1921
AGUUCUTCUUAUUCAGCACGAAC
2799
1899-1921





AD-565904.3
CGUGCUGAAUAAGAAGAACAU
2539
1902-1922
AUGUUCTUCUUAUUCAGCACGAA
2800
1900-1922





AD-1069878.1
GUGCUGAAUAAGAAGAACAAU
2540
1903-1923
AUUGUUCUUCUUAUUCAGCACGA
2801
1901-1923





AD-565906.1
UGCUGAAUAAGAAGAACAAAU
2541
1904-1924
AUUUGUTCUUCUUAUUCAGCACG
2802
1902-1924





AD-565907.1
GCUGAAUAAGAAGAACAAACU
2542
1905-1925
AGUUUGTUCUUCUUAUUCAGCAC
2803
1903-1925





AD-1069879.1
CUGAAUAAGAAGAACAAACUU
2543
1906-1926
AAGUUUGUUCUUCUUAUUCAGCA
2804
1904-1926





AD-565909.1
UGAAUAAGAAGAACAAACUGU
2544
1907-1927
ACAGUUTGUUCUUCUUAUUCAGC
2805
1905-1927





AD-565910.1
GAAUAAGAAGAACAAACUGAU
2545
1908-1928
AUCAGUTUGUUCUUCUUAUUCAG
2806
1906-1928





AD-565911.1
AAUAAGAAGAACAAACUGACU
2546
1909-1929
AGUCAGTUUGUUCUUCUUAUUCA
2807
1907-1929





AD-1069880.1
AUAAGAAGAACAAACUGACGU
2547
1910-1930
ACGUCAGUUUGUUCUUCUUAUUC
2808
1908-1930





AD-565913.1
UAAGAAGAACAAACUGACGCU
2548
1911-1931
AGCGUCAGUUUGUUCUUCUUAUU
2809
1909-1931





AD-1069881.1
AAGAAGAACAAACUGACGCAU
2549
1912-1932
AUGCGUCAGUUUGUUCUUCUUAU
2810
1910-1932





AD-565915.1
AGAAGAACAAACUGACGCAGU
2550
1913-1933
ACUGCGTCAGUUUGUUCUUCUUA
2811
1911-1933





AD-1069882.1
GAAGAACAAACUGACGCAGAU
2551
1914-1934
AUCUGCGUCAGUUUGUUCUUCUU
2812
1912-1934





AD-1069883.1
AAGAACAAACUGACGCAGAGU
2552
1915-1935
ACUCUGCGUCAGUUUGUUCUUCU
2813
1913-1935





AD-1069884.1
AGAACAAACUGACGCAGAGUU
2553
1916-1936
AACUCUGCGUCAGUUUGUUCUUC
2814
1914-1936





AD-565919.1
GAACAAACUGACGCAGAGUAU
2554
1917-1937
AUACUCTGCGUCAGUUUGUUCUU
2815
1915-1937





AD-1069885.1
AACAAACUGACGCAGAGUAAU
2555
1918-1938
AUUACUCUGCGUCAGUUUGUUCU
2816
1916-1938





AD-565921.1
ACAAACUGACGCAGAGUAAGU
2556
1919-1939
ACUUACTCUGCGUCAGUUUGUUC
2817
1917-1939





AD-1069886.1
CAAACUGACGCAGAGUAAGAU
2557
1920-1940
AUCUUACUCUGCGUCAGUUUGUU
2818
1918-1940





AD-565923.1
AAACUGACGCAGAGUAAGAUU
2558
1921-1941
AAUCUUACUCUGCGUCAGUUUGU
2819
1919-1941





AD-565924.1
AACUGACGCAGAGUAAGAUCU
2559
1922-1942
AGAUCUTACUCUGCGUCAGUUUG
2820
1920-1942





AD-1069887.1
CUGACGCAGAGUAAGAUCUGU
2560
1924-1944
ACAGAUCUUACUCUGCGUCAGUU
2821
1922-1944





AD-565927.1
UGACGCAGAGUAAGAUCUGGU
2561
1925-1945
ACCAGATCUUACUCUGCGUCAGU
2822
1923-1945





AD-565928.1
GACGCAGAGUAAGAUCUGGGU
2562
1926-1946
ACCCAGAUCUUACUCUGCGUCAG
2823
1924-1946





AD-1069888.1
ACGCAGAGUAAGAUCUGGGAU
2563
1927-1947
AUCCCAGAUCUUACUCUGCGUCA
2824
1925-1947





AD-566379.1
UGAGCAUGUCGGACAAGAAAU
2564
2513-2533
AUUUCUTGUCCGACAUGCUCACA
2825
2511-2533





AD-566380.1
GAGCAUGUCGGACAAGAAAGU
2565
2514-2534
ACUUUCTUGUCCGACAUGCUCAC
2826
2512-2534





AD-1069889.1
AGCAUGUCGGACAAGAAAGGU
2566
2515-2535
ACCUUUCUUGUCCGACAUGCUCA
2827
2513-2535





AD-566382.1
GCAUGUCGGACAAGAAAGGGU
2567
2516-2536
ACCCUUTCUUGUCCGACAUGCUC
2828
2514-2536





AD-566383.2
CAUGUCGGACAAGAAAGGGAU
2568
2517-2537
AUCCCUTUCUUGUCCGACAUGCU
2829
2515-2537





AD-566384.2
AUGUCGGACAAGAAAGGGAUU
2569
2518-2538
AAUCCCTUUCUUGUCCGACAUGC
2830
2516-2538





AD-1069890.1
UGUCGGACAAGAAAGGGAUCU
2570
2519-2539
AGAUCCCUUUCUUGUCCGACAUG
2831
2517-2539





AD-1069891.1
GUCGGACAAGAAAGGGAUCUU
2571
2520-2540
AAGAUCCCUUUCUUGUCCGACAU
2832
2518-2540





AD-1069892.1
UCGGACAAGAAAGGGAUCUGU
2572
2521-2541
ACAGAUCCCUUUCUUGUCCGACA
2833
2519-2541





AD-566388.2
CGGACAAGAAAGGGAUCUGUU
2573
2522-2542
AACAGATCCCUUUCUUGUCCGAC
2834
2520-2542





AD-566389.1
GGACAAGAAAGGGAUCUGUGU
2574
2523-2543
ACACAGAUCCCUUUCUUGUCCGA
2835
2521-2543





AD-1069893.1
GACAAGAAAGGGAUCUGUGUU
2575
2524-2544
AACACAGAUCCCUUUCUUGUCCG
2836
2522-2544





AD-566391.1
ACAAGAAAGGGAUCUGUGUGU
2576
2525-2545
ACACACAGAUCCCUUUCUUGUCC
2837
2523-2545





AD-1069894.1
CAAGAAAGGGAUCUGUGUGGU
2577
2526-2546
ACCACACAGAUCCCUUUCUUGUC
2838
2524-2546





AD-566393.1
AAGAAAGGGAUCUGUGUGGCU
2578
2527-2547
AGCCACACAGAUCCCUUUCUUGU
2839
2525-2547





AD-566395.1
GAAAGGGAUCUGUGUGGCAGU
2579
2529-2549
ACUGCCACACAGAUCCCUUUCUU
2840
2527-2549





AD-1069896.1
AAAGGGAUCUGUGUGGCAGAU
2580
2530-2550
AUCUGCCACACAGAUCCCUUUCU
2841
2528-2550





AD-1069897.1
AAGGGAUCUGUGUGGCAGACU
2581
2531-2551
AGUCUGCCACACAGAUCCCUUUC
2842
2529-2551





AD-1069898.1
AGGGAUCUGUGUGGCAGACCU
2582
2532-2552
AGGUCUGCCACACAGAUCCCUUU
2843
2530-2552





AD-1069899.1
GGGAUCUGUGUGGCAGACCCU
2583
2533-2553
AGGGUCTGCCACACAGAUCCCUU
2844
2531-2553





AD-566475.1
GAAAUCCGAGCCGUUCUCUAU
2584
2629-2649
AUAGAGAACGGCUCGGAUUUCCA
2845
2627-2649





AD-1069900.1
AAAUCCGAGCCGUUCUCUACU
2585
2630-2650
AGUAGAGAACGGCUCGGAUUUCC
2846
2628-2650





AD-566477.1
AAUCCGAGCCGUUCUCUACAU
2586
2631-2651
AUGUAGAGAACGGCUCGGAUUUC
2847
2629-2651





AD-1069901.1
AUCCGAGCCGUUCUCUACAAU
2587
2632-2652
AUUGUAGAGAACGGCUCGGAUUU
2848
2630-2652





AD-566483.1
AGCCGUUCUCUACAAUUACCU
2588
2637-2657
AGGUAATUGUAGAGAACGGCUCG
2849
2635-2657





AD-566484.1
GCCGUUCUCUACAAUUACCGU
2589
2638-2658
ACGGUAAUUGUAGAGAACGGCUC
2850
2636-2658





AD-566485.2
CCGUUCUCUACAAUUACCGGU
2590
2639-2659
ACCGGUAAUUGUAGAGAACGGCU
2851
2637-2659





AD-566486.1
CGUUCUCUACAAUUACCGGCU
2591
2640-2660
AGCCGGTAAUUGUAGAGAACGGC
2852
2638-2660





AD-1069902.1
GUUCUCUACAAUUACCGGCAU
2592
2641-2661
AUGCCGGUAAUUGUAGAGAACGG
2853
2639-2661





AD-1069903.1
UUCUCUACAAUUACCGGCAGU
2593
2642-2662
ACUGCCGGUAAUUGUAGAGAACG
2854
2640-2662





AD-1069904.1
UCUCUACAAUUACCGGCAGAU
2594
2643-2663
AUCUGCCGGUAAUUGUAGAGAAC
2855
2641-2663





AD-1069905.1
GGCUGACCGCCUACGUGGUCU
2595
3323-3343
AGACCACGUAGGCGGUCAGCCAG
2856
3321-3343





AD-567054.1
GCUGACCGCCUACGUGGUCAU
2596
3324-3344
AUGACCACGUAGGCGGUCAGCCA
2857
3322-3344





AD-1069906.1
CUGACCGCCUACGUGGUCAAU
2597
3325-3345
AUUGACCACGUAGGCGGUCAGCC
2858
3323-3345





AD-1069907.1
UGACCGCCUACGUGGUCAAGU
2598
3326-3346
ACUUGACCACGUAGGCGGUCAGC
2859
3324-3346





AD-567057.1
GACCGCCUACGUGGUCAAGGU
2599
3327-3347
ACCUUGACCACGUAGGCGGUCAG
2860
3325-3347





AD-1069908.1
ACCGCCUACGUGGUCAAGGUU
2600
3328-3348
AACCUUGACCACGUAGGCGGUCA
2861
3326-3348





AD-567059.1
CCGCCUACGUGGUCAAGGUCU
2601
3329-3349
AGACCUTGACCACGUAGGCGGUC
2862
3327-3349





AD-567060.1
CGCCUACGUGGUCAAGGUCUU
2602
3330-3350
AAGACCTUGACCACGUAGGCGGU
2863
3328-3350





AD-1069909.1
GCCUACGUGGUCAAGGUCUUU
2603
3331-3351
AAAGACCUUGACCACGUAGGCGG
2864
3329-3351





AD-1069910.1
CCUACGUGGUCAAGGUCUUCU
2604
3332-3352
AGAAGACCUUGACCACGUAGGCG
2865
3330-3352





AD-567063.4
CUACGUGGUCAAGGUCUUCUU
2605
3333-3353
AAGAAGACCUUGACCACGUAGGC
2866
3331-3353





AD-1069911.1
UACGUGGUCAAGGUCUUCUCU
2606
3334-3354
AGAGAAGACCUUGACCACGUAGG
2867
3332-3354





AD-567065.1
ACGUGGUCAAGGUCUUCUCUU
2607
3335-3355
AAGAGAAGACCUUGACCACGUAG
2868
3333-3355





AD-567066.4
CGUGGUCAAGGUCUUCUCUCU
2608
3336-3356
AGAGAGAAGACCUUGACCACGUA
2869
3334-3356





AD-1069912.1
GUGGUCAAGGUCUUCUCUCUU
2609
3337-3357
AAGAGAGAAGACCUUGACCACGU
2870
3335-3357





AD-567068.1
UGGUCAAGGUCUUCUCUCUGU
2610
3338-3358
ACAGAGAGAAGACCUUGACCACG
2871
3336-3358





AD-1069913.1
GGUCAAGGUCUUCUCUCUGGU
2611
3339-3359
ACCAGAGAGAAGACCUUGACCAC
2872
3337-3359





AD-567070.1
GUCAAGGUCUUCUCUCUGGCU
2612
3340-3360
AGCCAGAGAGAAGACCUUGACCA
2873
3338-3360





AD-1069914.1
UCAAGGUCUUCUCUCUGGCUU
2613
3341-3361
AAGCCAGAGAGAAGACCUUGACC
2874
3339-3361





AD-567072.1
CAAGGUCUUCUCUCUGGCUGU
2614
3342-3362
ACAGCCAGAGAGAAGACCUUGAC
2875
3340-3362





AD-1069915.1
AAGGUCUUCUCUCUGGCUGUU
2615
3343-3363
AACAGCCAGAGAGAAGACCUUGA
2876
3341-3363





AD-1069916.1
AGGUCUUCUCUCUGGCUGUCU
2616
3344-3364
AGACAGCCAGAGAGAAGACCUUG
2877
3342-3364





AD-1069917.1
GGUCUUCUCUCUGGCUGUCAU
2617
3345-3365
AUGACAGCCAGAGAGAAGACCUU
2878
3343-3365





AD-567076.1
GUCUUCUCUCUGGCUGUCAAU
2618
3346-3366
AUUGACAGCCAGAGAGAAGACCU
2879
3344-3366





AD-1069918.1
UCUUCUCUCUGGCUGUCAACU
2619
3347-3367
AGUUGACAGCCAGAGAGAAGACC
2880
3345-3367





AD-567294.1
UAAAGCAGGAGACUUCCUUGU
2620
3603-3623
ACAAGGAAGUCUCCUGCUUUAGU
2881
3601-3623





AD-1069919.1
AAAGCAGGAGACUUCCUUGAU
2621
3604-3624
AUCAAGGAAGUCUCCUGCUUUAG
2882
3602-3624





AD-1069920.1
AAGCAGGAGACUUCCUUGAAU
2622
3605-3625
AUUCAAGGAAGUCUCCUGCUUUA
2883
3603-3625





AD-567297.1
AGCAGGAGACUUCCUUGAAGU
2623
3606-3626
ACUUCAAGGAAGUCUCCUGCUUU
2884
3604-3626





AD-567300.1
AGGAGACUUCCUUGAAGCCAU
2624
3609-3629
AUGGCUTCAAGGAAGUCUCCUGC
2885
3607-3629





AD-567301.1
GGAGACUUCCUUGAAGCCAAU
2625
3610-3630
AUUGGCTUCAAGGAAGUCUCCUG
2886
3608-3630





AD-1069922.1
GAGACUUCCUUGAAGCCAACU
2626
3611-3631
AGUUGGCUUCAAGGAAGUCUCCU
2887
3609-3631





AD-1069923.1
AGACUUCCUUGAAGCCAACUU
2627
3612-3632
AAGUUGGCUUCAAGGAAGUCUCC
2888
3610-3632





AD-1069924.1
GACUUCCUUGAAGCCAACUAU
2628
3613-3633
AUAGUUGGCUUCAAGGAAGUCUC
2889
3611-3633





AD-567305.1
ACUUCCUUGAAGCCAACUACU
2629
3614-3634
AGUAGUTGGCUUCAAGGAAGUCU
2890
3612-3634





AD-567306.1
CUUCCUUGAAGCCAACUACAU
2630
3615-3635
AUGUAGTUGGCUUCAAGGAAGUC
2891
3613-3635





AD-567308.1
UCCUUGAAGCCAACUACAUGU
2631
3617-3637
ACAUGUAGUUGGCUUCAAGGAAG
2892
3615-3637





AD-567309.1
CCUUGAAGCCAACUACAUGAU
2632
3618-3638
AUCAUGTAGUUGGCUUCAAGGAA
2893
3616-3638





AD-1069925.1
CUUGAAGCCAACUACAUGAAU
2633
3619-3639
AUUCAUGUAGUUGGCUUCAAGGA
2894
3617-3639





AD-567311.1
UUGAAGCCAACUACAUGAACU
2634
3620-3640
AGUUCATGUAGUUGGCUUCAAGG
2895
3618-3640





AD-567312.1
UGAAGCCAACUACAUGAACCU
2635
3621-3641
AGGUUCAUGUAGUUGGCUUCAAG
2896
3619-3641





AD-1069926.1
GAAGCCAACUACAUGAACCUU
2636
3622-3642
AAGGUUCAUGUAGUUGGCUUCAA
2897
3620-3642





AD-567314.2
AAGCCAACUACAUGAACCUAU
2637
3623-3643
AUAGGUTCAUGUAGUUGGCUUCA
2898
3621-3643





AD-567315.6
AGCCAACUACAUGAACCUACU
2638
3624-3644
AGUAGGTUCAUGUAGUUGGCUUC
2899
3622-3644





AD-1069927.1
GCCAACUACAUGAACCUACAU
2639
3625-3645
AUGUAGGUUCAUGUAGUUGGCUU
2900
3623-3645





AD-1069928.1
CCAACUACAUGAACCUACAGU
2640
3626-3646
ACUGUAGGUUCAUGUAGUUGGCU
2901
3624-3646





AD-567318.2
CAACUACAUGAACCUACAGAU
2641
3627-3647
AUCUGUAGGUUCAUGUAGUUGGC
2902
3625-3647





AD-567319.1
AACUACAUGAACCUACAGAGU
2642
3628-3648
ACUCUGTAGGUUCAUGUAGUUGG
2903
3626-3648





AD-1069929.1
ACUACAUGAACCUACAGAGAU
2643
3629-3649
AUCUCUGUAGGUUCAUGUAGUUG
2904
3627-3649





AD-567321.1
CUACAUGAACCUACAGAGAUU
2644
3630-3650
AAUCUCTGUAGGUUCAUGUAGUU
2905
3628-3650





AD-1069930.1
UACAUGAACCUACAGAGAUCU
2645
3631-3651
AGAUCUCUGUAGGUUCAUGUAGU
2906
3629-3651





AD-567323.1
ACAUGAACCUACAGAGAUCCU
2646
3632-3652
AGGAUCTCUGUAGGUUCAUGUAG
2907
3630-3652





AD-1069931.1
CAUGAACCUACAGAGAUCCUU
2647
3633-3653
AAGGAUCUCUGUAGGUUCAUGUA
2908
3631-3653





AD-567325.1
AUGAACCUACAGAGAUCCUAU
2648
3634-3654
AUAGGATCUCUGUAGGUUCAUGU
2909
3632-3654





AD-567326.1
UGAACCUACAGAGAUCCUACU
2649
3635-3655
AGUAGGAUCUCUGUAGGUUCAUG
2910
3633-3655





AD-1069932.1
GAACCUACAGAGAUCCUACAU
2650
3636-3656
AUGUAGGAUCUCUGUAGGUUCAU
2911
3634-3656





AD-1069933.1
AACCUACAGAGAUCCUACACU
2651
3637-3657
AGUGUAGGAUCUCUGUAGGUUCA
2912
3635-3657





AD-567479.1
GGCCCUACUGCAGCUAAAAGU
2652
3807-3827
ACUUUUAGCUGCAGUAGGGCCAA
2913
3805-3827





AD-567480.1
GCCCUACUGCAGCUAAAAGAU
2653
3808-3828
AUCUUUTAGCUGCAGUAGGGCCA
2914
3806-3828





AD-567481.1
CCCUACUGCAGCUAAAAGACU
2654
3809-3829
AGUCUUTUAGCUGCAGUAGGGCC
2915
3807-3829





AD-567482.1
CCUACUGCAGCUAAAAGACUU
2655
3810-3830
AAGUCUTUUAGCUGCAGUAGGGC
2916
3808-3830





AD-1069934.1
UACUGCAGCUAAAAGACUUUU
2656
3812-3832
AAAAGUCUUUUAGCUGCAGUAGG
2917
3810-3832





AD-567485.1
ACUGCAGCUAAAAGACUUUGU
2657
3813-3833
ACAAAGTCUUUUAGCUGCAGUAG
2918
3811-3833





AD-1069935.1
CUGCAGCUAAAAGACUUUGAU
2658
3814-3834
AUCAAAGUCUUUUAGCUGCAGUA
2919
3812-3834





AD-567487.2
UGCAGCUAAAAGACUUUGACU
2659
3815-3835
AGUCAAAGUCUUUUAGCUGCAGU
2920
3813-3835





AD-567488.1
GCAGCUAAAAGACUUUGACUU
2660
3816-3836
AAGUCAAAGUCUUUUAGCUGCAG
2921
3814-3836





AD-567489.1
CAGCUAAAAGACUUUGACUUU
2661
3817-3837
AAAGUCAAAGUCUUUUAGCUGCA
2922
3815-3837





AD-1069936.1
AGCUAAAAGACUUUGACUUUU
2662
3818-3838
AAAAGUCAAAGUCUUUUAGCUGC
2923
3816-3838





AD-567491.1
GCUAAAAGACUUUGACUUUGU
2663
3819-3839
ACAAAGTCAAAGUCUUUUAGCUG
2924
3817-3839





AD-1069937.1
GUGCCUCCCGUCGUGCGUUGU
2664
3838-3858
ACAACGCACGACGGGAGGCACAA
2925
3836-3858





AD-1069938.1
UGCCUCCCGUCGUGCGUUGGU
2665
3839-3859
ACCAACGCACGACGGGAGGCACA
2926
3837-3859





AD-1069939.1
GCCUCCCGUCGUGCGUUGGCU
2666
3840-3860
AGCCAACGCACGACGGGAGGCAC
2927
3838-3860





AD-567513.1
CCUCCCGUCGUGCGUUGGCUU
2667
3841-3861
AAGCCAACGCACGACGGGAGGCA
2928
3839-3861





AD-567514.1
CUCCCGUCGUGCGUUGGCUCU
2668
3842-3862
AGAGCCAACGCACGACGGGAGGC
2929
3840-3862





AD-1069940.1
UCCCGUCGUGCGUUGGCUCAU
2669
3843-3863
AUGAGCCAACGCACGACGGGAGG
2930
3841-3863





AD-1069941.1
CCCGUCGUGCGUUGGCUCAAU
2670
3844-3864
AUUGAGCCAACGCACGACGGGAG
2931
3842-3864





AD-1069942.1
CCGUCGUGCGUUGGCUCAAUU
2671
3845-3865
AAUUGAGCCAACGCACGACGGGA
2932
3843-3865





AD-567518.1
CGUCGUGCGUUGGCUCAAUGU
2672
3846-3866
ACAUUGAGCCAACGCACGACGGG
2933
3844-3866





AD-1069943.1
GUCGUGCGUUGGCUCAAUGAU
2673
3847-3867
AUCAUUGAGCCAACGCACGACGG
2934
3845-3867





AD-567521.4
CGUGCGUUGGCUCAAUGAACU
2674
3849-3869
AGUUCATUGAGCCAACGCACGAC
2935
3847-3869





AD-1069944.1
UGCGUUGGCUCAAUGAACAGU
2675
3851-3871
ACUGUUCAUUGAGCCAACGCACG
2936
3849-3871





AD-567524.1
GCGUUGGCUCAAUGAACAGAU
2676
3852-3872
AUCUGUTCAUUGAGCCAACGCAC
2937
3850-3872





AD-567525.1
CGUUGGCUCAAUGAACAGAGU
2677
3853-3873
ACUCUGTUCAUUGAGCCAACGCA
2938
3851-3873





AD-1069945.1
GUUGGCUCAAUGAACAGAGAU
2678
3854-3874
AUCUCUGUUCAUUGAGCCAACGC
2939
3852-3874





AD-567527.1
UUGGCUCAAUGAACAGAGAUU
2679
3855-3875
AAUCUCTGUUCAUUGAGCCAACG
2940
3853-3875





AD-1069946.1
UGGCUCAAUGAACAGAGAUAU
2680
3856-3876
AUAUCUCUGUUCAUUGAGCCAAC
2941
3854-3876





AD-567529.1
GGCUCAAUGAACAGAGAUACU
2681
3857-3877
AGUAUCTCUGUUCAUUGAGCCAA
2942
3855-3877





AD-1069947.1
GCUCAAUGAACAGAGAUACUU
2682
3858-3878
AAGUAUCUCUGUUCAUUGAGCCA
2943
3856-3878





AD-567531.1
CUCAAUGAACAGAGAUACUAU
2683
3859-3879
AUAGUATCUCUGUUCAUUGAGCC
2944
3857-3879





AD-567532.1
UCAAUGAACAGAGAUACUACU
2684
3860-3880
AGUAGUAUCUCUGUUCAUUGAGC
2945
3858-3880





AD-567533.1
CAAUGAACAGAGAUACUACGU
2685
3861-3881
ACGUAGTAUCUCUGUUCAUUGAG
2946
3859-3881





AD-1069948.1
AAUGAACAGAGAUACUACGGU
2686
3862-3882
ACCGUAGUAUCUCUGUUCAUUGA
2947
3860-3882





AD-567535.1
AUGAACAGAGAUACUACGGUU
2687
3863-3883
AACCGUAGUAUCUCUGUUCAUUG
2948
3861-3883





AD-568149.1
GAGCAGUCAAGGUCUACGCCU
2688
4517-4537
AGGCGUAGACCUUGACUGCUCCA
2949
4515-4537





AD-568150.1
AGCAGUCAAGGUCUACGCCUU
2689
4518-4538
AAGGCGTAGACCUUGACUGCUCC
2950
4516-4538





AD-1069949.1
GCAGUCAAGGUCUACGCCUAU
2690
4519-4539
AUAGGCGUAGACCUUGACUGCUC
2951
4517-4539





AD-1069950.1
CAGUCAAGGUCUACGCCUAUU
2691
4520-4540
AAUAGGCGUAGACCUUGACUGCU
2952
4518-4540





AD-1069951.1
AGUCAAGGUCUACGCCUAUUU
2692
4521-4541
AAAUAGGCGUAGACCUUGACUGC
2953
4519-4541





AD-1069952.1
GUCAAGGUCUACGCCUAUUAU
2693
4522-4542
AUAAUAGGCGUAGACCUUGACUG
2954
4520-4542





AD-568155.1
UCAAGGUCUACGCCUAUUACU
2694
4523-4543
AGUAAUAGGCGUAGACCUUGACU
2955
4521-4543





AD-568159.1
GGUCUACGCCUAUUACAACCU
2695
4527-4547
AGGUUGTAAUAGGCGUAGACCUU
2956
4525-4547





AD-1069953.1
GUCUACGCCUAUUACAACCUU
2696
4528-4548
AAGGUUGUAAUAGGCGUAGACCU
2957
4526-4548





AD-568161.2
UCUACGCCUAUUACAACCUGU
2697
4529-4549
ACAGGUTGUAAUAGGCGUAGACC
2958
4527-4549





AD-568162.1
CUACGCCUAUUACAACCUGGU
2698
4530-4550
ACCAGGTUGUAAUAGGCGUAGAC
2959
4528-4550





AD-1069954.1
UACGCCUAUUACAACCUGGAU
2699
4531-4551
AUCCAGGUUGUAAUAGGCGUAGA
2960
4529-4551





AD-1069955.1
ACGCCUAUUACAACCUGGAGU
2700
4532-4552
ACUCCAGGUUGUAAUAGGCGUAG
2961
4530-4552





AD-568165.1
CGCCUAUUACAACCUGGAGGU
2701
4533-4553
ACCUCCAGGUUGUAAUAGGCGUA
2962
4531-4553





AD-1069956.1
GCUGAGGAGAAUUGCUUCAUU
2702
4633-4653
AAUGAAGCAAUUCUCCUCAGCAC
2963
4631-4653





AD-568337.1
GCCAGGAGUGGACUAUGUGUU
2703
4707-4727
AACACATAGUCCACUCCUGGCUC
2964
4705-4727





AD-568338.1
CCAGGAGUGGACUAUGUGUAU
2704
4708-4728
AUACACAUAGUCCACUCCUGGCU
2965
4706-4728





AD-1069957.1
CAGGAGUGGACUAUGUGUACU
2705
4709-4729
AGUACACAUAGUCCACUCCUGGC
2966
4707-4729





AD-568340.1
AGGAGUGGACUAUGUGUACAU
2706
4710-4730
AUGUACACAUAGUCCACUCCUGG
2967
4708-4730





AD-1069958.1
GGAGUGGACUAUGUGUACAAU
2707
4711-4731
AUUGUACACAUAGUCCACUCCUG
2968
4709-4731





AD-568342.1
GAGUGGACUAUGUGUACAAGU
2708
4712-4732
ACUUGUACACAUAGUCCACUCCU
2969
4710-4732





AD-568343.4
AGUGGACUAUGUGUACAAGAU
2709
4713-4733
AUCUUGTACACAUAGUCCACUCC
2970
4711-4733





AD-1069959.1
GUGGACUAUGUGUACAAGACU
2710
4714-4734
AGUCUUGUACACAUAGUCCACUC
2971
4712-4734





AD-568345.2
UGGACUAUGUGUACAAGACCU
2711
4715-4735
AGGUCUTGUACACAUAGUCCACU
2972
4713-4735





AD-568348.1
ACUAUGUGUACAAGACCCGAU
2712
4718-4738
AUCGGGTCUUGUACACAUAGUCC
2973
4716-4738





AD-1069961.1
CUAUGUGUACAAGACCCGACU
2713
4719-4739
AGUCGGGUCUUGUACACAUAGUC
2974
4717-4739
















TABLE 23







Modified Sense and Antisense Strand Sequences of Complement Component C3 dsRNA Agents















SEQ

SEQ

SEQ




ID

ID

ID


Duplex Name
Sense Sequene 5’ to 3’
NO:
Antisense Sequence 5’ to 3’
NO:
mRNA Target Sequence
NO:





AD-564723.1
asgsagcgGfgUfAfCfcucuucaucuL96
2975
asGfsauga(Agn)gagguaCfcCfgcucusgsc
3236
GCAGAGCGGGUACCUCUUCAUCC
3497





AD-564724.1
gsasgcggGfuAfCfCfucuucauccuL96
2976
asGfsgaug(Agn)agagguAfcCfegcucsusg
3237
CAGAGCGGGUACCUCUUCAUCCA
3498





AD-1069838.1
asgscgggUfaCfCfUfcuucauccauL96
2977
asUfsggau(G2p)aagaggUfaCfecgcuscsu
3238
AGAGCGGGUACCUCUUCAUCCAG
3499





AD-564726.1
gscsggguAfcCfUfCfuucauccaguL96
2978
asCfsugga(Tgn)gaagagGfuAfeccgcsusc
3239
GAGCGGGUACCUCUUCAUCCAGA
3500





AD-564727.3
csgsgguaCfcUfCfUfucauccagauL96
2979
asUfscugg(Agn)ugaagaGfgUfacccgscsu
3240
AGCGGGUACCUCUUCAUCCAGAC
3501





AD-1069839.1
gsgsguacCfuCfUfUfcauccagacuL96
2980
asGfsucug(G2p)augaagAfgGfuacccsgsc
3241
GCGGGUACCUCUUCAUCCAGACA
3502





AD-1069840.1
gsgsuaccUfcUfUfCfauccagacauL96
2981
asUfsgucu(G2p)gaugaaGfaGfguaccscsg
3242
CGGGUACCUCUUCAUCCAGACAG
3503





AD-564730.3
gsusaccuCfuUfCfAfuccagacaguL96
2982
asCfsuguc(Tgn)ggaugaAfgAfgguacscsc
3243
GGGUACCUCUUCAUCCAGACAGA
3504





AD-1069841.1
usasccucUfuCfAfUfccagacagauL96
2983
asUfscugu(C2p)uggaugAfaGfagguascsc
3244
GGUACCUCUUCAUCCAGACAGAC
3505





AD-564732.1
ascscucuUfcAfUfCfcagacagacuL96
2984
asGfsucug(Tgn)cuggauGfaAfgaggusasc
3245
GUACCUCUUCAUCCAGACAGACA
3506





AD-1069842.1
cscsucuuCfaUfCfCfagacagacauL96
2985
asUfsgucu(G2p)ucuggaUfgAfagaggsusa
3246
UACCUCUUCAUCCAGACAGACAA
3507





AD-564734.1
csuscuucAfuCfCfAfgacagacaauL96
2986
asUfsuguc(Tgn)gucuggAfuGfaagagsgsu
3247
ACCUCUUCAUCCAGACAGACAAG
3508





AD-1069843.1
uscsuucaUfcCfAfGfacagacaaguL96
2987
asCfsuugu(C2p)ugucugGfaUfgaagasgsg
3248
CCUCUUCAUCCAGACAGACAAGA
3509





AD-564736.1
csusucauCfcAfGfAfcagacaagauL96
2988
asUfscuug(Tgn)cugucuGfgAfugaagsasg
3249
CUCUUCAUCCAGACAGACAAGAC
3510





AD-1069844.1
ususcaucCfaGfAfCfagacaagacuL96
2989
asGfsucuu(G2p)ucugucUfgGfaugaasgsa
3250
UCUUCAUCCAGACAGACAAGACC
3511





AD-564738.1
uscsauccAfgAfCfAfgacaagaccuL96
2990
asGfsgucu(Tgn)gucuguCfuGfgaugasasg
3251
CUUCAUCCAGACAGACAAGACCA
3512





AD-564739.2
csasuccaGfaCfAfGfacaagaccauL96
2991
asUfsgguc(Tgn)ugucugUfcUfggaugsasa
3252
UUCAUCCAGACAGACAAGACCAU
3513





AD-1069845.1
asusccagAfcAfGfAfcaagaccauuL96
2992
asAfsuggu(C2p)uugucuGfuCfuggausgsa
3253
UCAUCCAGACAGACAAGACCAUC
3514





AD-564741.1
uscscagaCfaGfAfCfaagaccaucuL96
2993
asGfsaugg(Tgn)cuugucUfgUfcuggasusg
3254
CAUCCAGACAGACAAGACCAUCU
3515





AD-1069846.1
cscsagacAfgAfCfAfagaccaucuuL96
2994
asAfsgaug(G2p)ucuuguCfuGfucuggsasu
3255
AUCCAGACAGACAAGACCAUCUA
3516





AD-1069847.1
csasgacaGfaCfAfAfgaccaucuauL96
2995
asUfsagau(G2p)gucuugUfcUfgucugsgsa
3256
UCCAGACAGACAAGACCAUCUAC
3517





AD-564745.3
gsascagaCfaAfGfAfccaucuacauL96
2996
asUfsguag(Agn)uggucuUfgUfcugucsusg
3257
CAGACAGACAAGACCAUCUACAC
3518





AD-564747.1
csasgacaAfgAfCfCfaucuacaccuL96
2997
asGfsgugu(Agn)gaugguCfuUfgucugsusc
3258
GACAGACAAGACCAUCUACACCC
3519





AD-1069850.1
ascsaagaCfcAfUfCfuacaccccuuL96
2998
asAfsgggg(Tgn)guagauGfgUfcuuguscsu
3259
AGACAAGACCAUCUACACCCCUG
3520





AD-1069851.1
gsgsccagUfgGfAfAfgauccgagcuL96
2999
asGfscucg(G2p)aucuucCfaCfuggccscsa
3260
UGGGCCAGUGGAAGAUCCGAGCC
3521





AD-1069852.1
gscscaguGfgAfAfGfauccgagccuL96
3000
asGfsgcuc(G2p)gaucuuCfcAfcuggcscsc
3261
GGGCCAGUGGAAGAUCCGAGCCU
3522





AD-1069853.1
cscsagugGfaAfGfAfuccgagccuuL96
3001
asAfsggcu(C2p)ggaucuUfcCfacuggscsc
3262
GGCCAGUGGAAGAUCCGAGCCUA
3523





AD-564925.1
csasguggAfaGfAfUfccgagccuauL96
3002
asUfsaggc(Tgn)cggaucUfuCfcacugsgsc
3263
GCCAGUGGAAGAUCCGAGCCUAC
3524





AD-1069854.1
asgsuggaAfgAfUfCfcgagccuacuL96
3003
asGfsuagg(C2p)ucggauCfuUfccacusgsg
3264
CCAGUGGAAGAUCCGAGCCUACU
3525





AD-1069855.1
gsusggaaGfaUfCfCfgagccuacuuL96
3004
asAfsguag(G2p)cucggaUfcUfuccacsusg
3265
CAGUGGAAGAUCCGAGCCUACUA
3526





AD-1069856.1
usgsgaagAfuCfCfGfagccuacuauL96
3005
asUfsagua(G2p)gcucggAfuCfuuccascsu
3266
AGUGGAAGAUCCGAGCCUACUAU
3527





AD-564929.1
gsgsaagaUfcCfGfAfgccuacuauuL96
3006
asAfsuagu(Agn)ggcucgGfaUfcuuccsasc
3267
GUGGAAGAUCCGAGCCUACUAUG
3528





AD-564930.1
gsasagauCfcGfAfGfccuacuauguL96
3007
asCfsauag(Tgn)aggcucGfgAfucuucscsa
3268
UGGAAGAUCCGAGCCUACUAUGA
3529





AD-1069857.1
asasgaucCfgAfGfCfcuacuaugauL96
3008
asUfscaua(G2p)uaggcuCfgGfaucuuscsc
3269
GGAAGAUCCGAGCCUACUAUGAA
3530





AD-564934.1
asusccgaGfcCfUfAfcuaugaaaauL96
3009
asUfsuuuc(Agn)uaguagGfcUfcggauscsu
3270
AGAUCCGAGCCUACUAUGAAAAC
3531





AD-1069858.1
uscscgagCfcUfAfCfuaugaaaacuL96
3010
asGfsuuuu(C2p)auaguaGfgCfucggasusc
3271
GAUCCGAGCCUACUAUGAAAACU
3532





AD-564936.1
cscsgagcCfuAfCfUfaugaaaacuuL96
3011
asAfsguuu(Tgn)cauaguAfgGfcucggsasu
3272
AUCCGAGCCUACUAUGAAAACUC
3533





AD-564937.1
csgsagccUfaCfUfAfugaaaacucuL96
3012
asGfsaguu(Tgn)ucauagUfaGfgcucgsgsa
3273
UCCGAGCCUACUAUGAAAACUCA
3534





AD-564938.1
gsasgccuAfcUfAfUfgaaaacucauL96
3013
asUfsgagu(Tgn)uucauaGfuAfggcucsgsg
3274
CCGAGCCUACUAUGAAAACUCAC
3535





AD-1069859.1
gscscuacUfaUfGfAfaaacucaccuL96
3014
asGfsguga(G2p)uuuucaUfaGfuaggcsusc
3275
GAGCCUACUAUGAAAACUCACCA
3536





AD-564941.1
cscsuacuAfuGfAfAfaacucaccauL96
3015
asUfsggug(Agn)guuuucAfuAfguaggscsu
3276
AGCCUACUAUGAAAACUCACCAC
3537





AD-1069860.1
csusacuaUfgAfAfAfacucaccacuL96
3016
asGfsuggu(G2p)aguuuuCfaUfaguagsgsc
3277
GCCUACUAUGAAAACUCACCACA
3538





AD-564943.1
usascuauGfaAfAfAfcucaccacauL96
3017
asUfsgugg(Tgn)gaguuuUfcAfuaguasgsg
3278
CCUACUAUGAAAACUCACCACAG
3539





AD-1069861.1
cscsuacaGfaGfAfAfauucuacuauL96
3018
asUfsagua(G2p)aauuucUfcUfguaggscsu
3279
AGCCUACAGAGAAAUUCUACUAC
3540





AD-565031.1
csusacagAfgAfAfAfuucuacuacuL96
3019
asGfsuagu(Agn)gaauuuCfuCfuguagsgsc
3280
GCCUACAGAGAAAUUCUACUACA
3541





AD-565032.1
usascagaGfaAfAfUfucuacuacauL96
3020
asUfsguag(Tgn)agaauuUfcUfcuguasgsg
3281
CCUACAGAGAAAUUCUACUACAU
3542





AD-1069862.1
ascsagagAfaAfUfUfcuacuacauuL96
3021
asAfsugua(G2p)uagaauUfuCfucugusasg
3282
CUACAGAGAAAUUCUACUACAUC
3543





AD-565034.1
csasgagaAfaUfUfCfuacuacaucuL96
3022
asGfsaugu(Agn)guagaaUfuUfcucugsusa
3283
UACAGAGAAAUUCUACUACAUCU
3544





AD-565035.1
asgsagaaAfuUfCfUfacuacaucuuL96
3023
asAfsgaug(Tgn)aguagaAfuUfucucusgsu
3284
ACAGAGAAAUUCUACUACAUCUA
3545





AD-1069863.1
gsasgaaaUfuCfUfAfcuacaucuauL96
3024
asUfsagau(G2p)uaguagAfaUfuucucsusg
3285
CAGAGAAAUUCUACUACAUCUAU
3546





AD-565037.1
asgsaaauUfcUfAfCfuacaucuauuL96
3025
asAfsuaga(Tgn)guaguaGfaAfuuucuscsu
3286
AGAGAAAUUCUACUACAUCUAUA
3547





AD-565038.1
gsasaauuCfuAfCfUfacaucuauauL96
3026
asUfsauag(Agn)uguaguAfgAfauuucsusc
3287
GAGAAAUUCUACUACAUCUAUAA
3548





AD-1069864.1
asasauucUfaCfUfAfcaucuauaauL96
3027
asUfsuaua(G2p)auguagUfaGfaauuuscsu
3288
AGAAAUUCUACUACAUCUAUAAC
3549





AD-565041.1
asusucuaCfuAfCfAfucuauaacguL96
3028
asCfsguua(Tgn)agauguAfgUfagaaususu
3289
AAAUUCUACUACAUCUAUAACGA
3550





AD-565042.1
ususcuacUfaCfAfUfcuauaacgauL96
3029
asUfscguu(Agn)uagaugUfaGfuagaasusu
3290
AAUUCUACUACAUCUAUAACGAG
3551





AD-565043.1
uscsuacuAfcAfUfCfuauaacgaguL96
3030
asCfsucgu(Tgn)auagauGfuAfguagasasu
3291
AUUCUACUACAUCUAUAACGAGA
3552





AD-565044.1
csusacuaCfaUfCfUfauaacgagauL96
3031
asUfscucg(Tgn)uauagaUfgUfaguagsasa
3292
UUCUACUACAUCUAUAACGAGAA
3553





AD-1069865.1
usascuacAfuCfUfAfuaacgagaauL96
3032
asUfsucuc(G2p)uuauagAfuGfuaguasgsa
3293
UCUACUACAUCUAUAACGAGAAG
3554





AD-1069866.1
ascsuacaUfcUfAfUfaacgagaaguL96
3033
asCfsuucu(C2p)guuauaGfaUfguagusasg
3294
CUACUACAUCUAUAACGAGAAGG
3555





AD-565047.1
csusacauCfuAfUfAfacgagaagguL96
3034
asCfscuuc(Tgn)cguuauAfgAfuguagsusa
3295
UACUACAUCUAUAACGAGAAGGG
3556





AD-1069867.1
usascaucUfaUfAfAfcgagaaggguL96
3035
asCfsccuu(C2p)ucguuaUfaGfauguasgsu
3296
ACUACAUCUAUAACGAGAAGGGC
3557





AD-565049.1
ascsaucuAfuAfAfCfgagaagggcuL96
3036
asGfscccu(Tgn)cucguuAfuAfgaugusasg
3297
CUACAUCUAUAACGAGAAGGGCC
3558





AD-565050.1
csasucuaUfaAfCfGfagaagggccuL96
3037
asGfsgccc(Tgn)ucucguUfaUfagaugsusa
3298
UACAUCUAUAACGAGAAGGGCCU
3559





AD-565274.1
cscsucucCfcUfAfCfcagauccacuL96
3038
asGfsugga(Tgn)cugguaGfgGfagaggsusc
3299
GACCUCUCCCUACCAGAUCCACU
3560





AD-565275.1
csuscuccCfuAfCfCfagauccacuuL96
3039
asAfsgugg(Agn)ucugguAfgGfgagagsgsu
3300
ACCUCUCCCUACCAGAUCCACUU
3561





AD-1069868.1
uscsucccUfaCfCfAfgauccacuuuL96
3040
asAfsagug(G2p)aucuggUfaGfggagasgsg
3301
CCUCUCCCUACCAGAUCCACUUC
3562





AD-1069869.1
csuscccuAfcCfAfGfauccacuucuL96
3041
asGfsaagu(G2p)gaucugGfuAfgggagsasg
3302
CUCUCCCUACCAGAUCCACUUCA
3563





AD-565278.2
uscsccuaCfcAfGfAfuccacuucauL96
3042
asUfsgaag(Tgn)ggaucuGfgUfagggasgsa
3303
UCUCCCUACCAGAUCCACUUCAC
3564





AD-1069870.1
cscscuacCfaGfAfUfccacuucacuL96
3043
asGfsugaa(G2p)uggaucUfgGfuagggsasg
3304
CUCCCUACCAGAUCCACUUCACC
3565





AD-565280.1
cscsuaccAfgAfUfCfcacuucaccuL96
3044
asGfsguga(Agn)guggauCfuGfguaggsgsa
3305
UCCCUACCAGAUCCACUUCACCA
3566





AD-565281.3
csusaccaGfaUfCfCfacuucaccauL96
3045
asUfsggug(Agn)aguggaUfcUfgguagsgsg
3306
CCCUACCAGAUCCACUUCACCAA
3567





AD-1069871.1
usasccagAfuCfCfAfcuucaccaauL96
3046
asUfsuggu(G2p)aaguggAfuCfugguasgsg
3307
CCUACCAGAUCCACUUCACCAAG
3568





AD-565283.1
ascscagaUfcCfAfCfuucaccaaguL96
3047
asCfsuugg(Tgn)gaagugGfaUfcuggusasg
3308
CUACCAGAUCCACUUCACCAAGA
3569





AD-1069872.1
cscsagauCfcAfCfUfucaccaagauL96
3048
asUfscuug(G2p)ugaaguGfgAfucuggsusa
3309
UACCAGAUCCACUUCACCAAGAC
3570





AD-1069873.1
csasgaucCfaCfUfUfcaccaagacuL96
3049
asGfsucuu(G2p)gugaagUfgGfaucugsgsu
3310
ACCAGAUCCACUUCACCAAGACA
3571





AD-565286.1
asgsauccAfcUfUfCfaccaagacauL96
3050
asUfsgucu(Tgn)ggugaaGfuGfgaucusgsg
3311
CCAGAUCCACUUCACCAAGACAC
3572





AD-565287.1
gsasuccaCfuUfCfAfccaagacacuL96
3051
asGfsuguc(Tgn)uggugaAfgUfggaucsusg
3312
CAGAUCCACUUCACCAAGACACC
3573





AD-1069874.1
asusccacUfuCfAfCfcaagacaccuL96
3052
asGfsgugu(C2p)uuggugAfaGfuggauscsu
3313
AGAUCCACUUCACCAAGACACCC
3574





AD-1069875.1
ususugacCfuCfAfUfgguguucguuL96
3053
asAfscgaa(C2p)accaugAfgGfucaaasgsg
3314
CCUUUGACCUCAUGGUGUUCGUG
3575





AD-565335.1
usgsaccuCfaUfGfGfuguucgugauL96
3054
asUfscacg(Agn)acaccaUfgAfggucasasa
3315
UUUGACCUCAUGGUGUUCGUGAC
3576





AD-1069876.1
asgsggcgUfgUfUfCfgugcugaauuL96
3055
asAfsuuca(G2p)cacgaaCfaCfgcccususg
3316
CAAGGGCGUGUUCGUGCUGAAUA
3577





AD-565895.1
gsgsgcguGfuUfCfGfugcugaauauL96
3056
asUfsauuc(Agn)gcacgaAfcAfcgcccsusu
3317
AAGGGCGUGUUCGUGCUGAAUAA
3578





AD-1069877.1
gsgscgugUfuCfGfUfgcugaauaauL96
3057
asUfsuauu(C2p)agcacgAfaCfacgccscsu
3318
AGGGCGUGUUCGUGCUGAAUAAG
3579





AD-565897.1
gscsguguUfcGfUfGfcugaauaaguL96
3058
asCfsuuau(Tgn)cagcacGfaAfcacgcscsc
3319
GGGCGUGUUCGUGCUGAAUAAGA
3580





AD-565899.1
gsusguucGfuGfCfUfgaauaagaauL96
3059
asUfsucuu(Agn)uucagcAfcGfaacacsgsc
3320
GCGUGUUCGUGCUGAAUAAGAAG
3581





AD-565903.1
uscsgugcUfgAfAfUfaagaagaacuL96
3060
asGfsuucu(Tgn)cuuauuCfaGfcacgasasc
3321
GUUCGUGCUGAAUAAGAAGAACA
3582





AD-565904.3
csgsugcuGfaAfUfAfagaagaacauL96
3061
asUfsguuc(Tgn)ucuuauUfcAfgcacgsasa
3322
UUCGUGCUGAAUAAGAAGAACAA
3583





AD-1069878.1
gsusgcugAfaUfAfAfgaagaacaauL96
3062
asUfsuguu(C2p)uucuuaUfuCfagcacsgsa
3323
UCGUGCUGAAUAAGAAGAACAAA
3584





AD-565906.1
usgscugaAfuAfAfGfaagaacaaauL96
3063
asUfsuugu(Tgn)cuucuuAfuUfcagcascsg
3324
CGUGCUGAAUAAGAAGAACAAAC
3585





AD-565907.1
gscsugaaUfaAfGfAfagaacaaacuL96
3064
asGfsuuug(Tgn)ucuucuUfaUfucagcsasc
3325
GUGCUGAAUAAGAAGAACAAACU
3586





AD-1069879.1
csusgaauAfaGfAfAfgaacaaacuuL96
3065
asAfsguuu(G2p)uucuucUfuAfuucagscsa
3326
UGCUGAAUAAGAAGAACAAACUG
3587





AD-565909.1
usgsaauaAfgAfAfGfaacaaacuguL96
3066
asCfsaguu(Tgn)guucuuCfuUfauucasgsc
3327
GCUGAAUAAGAAGAACAAACUGA
3588





AD-565910.1
gsasauaaGfaAfGfAfacaaacugauL96
3067
asUfscagu(Tgn)uguucuUfcUfuauucsasg
3328
CUGAAUAAGAAGAACAAACUGAC
3589





AD-565911.1
asasuaagAfaGfAfAfcaaacugacuL96
3068
asGfsucag(Tgn)uuguucUfuCfuuauuscsa
3329
UGAAUAAGAAGAACAAACUGACG
3590





AD-1069880.1
asusaagaAfgAfAfCfaaacugacguL96
3069
asCfsguca(G2p)uuuguuCfuUfcuuaususc
3330
GAAUAAGAAGAACAAACUGACGC
3591





AD-565913.1
usasagaaGfaAfCfAfaacugacgcuL96
3070
asGfscguc(Agn)guuuguUfcUfucuuasusu
3331
AAUAAGAAGAACAAACUGACGCA
3592





AD-1069881.1
asasgaagAfaCfAfAfacugacgcauL96
3071
asUfsgcgu(C2p)aguuugUfuCfuucuusasu
3332
AUAAGAAGAACAAACUGACGCAG
3593





AD-565915.1
asgsaagaAfcAfAfAfcugacgcaguL96
3072
asCfsugcg(Tgn)caguuuGfuUfcuucususa
3333
UAAGAAGAACAAACUGACGCAGA
3594





AD-1069882.1
gsasagaaCfaAfAfCfugacgcagauL96
3073
asUfscugc(G2p)ucaguuUfgUfucuucsusu
3334
AAGAAGAACAAACUGACGCAGAG
3595





AD-1069883.1
asasgaacAfaAfCfUfgacgcagaguL96
3074
asCfsucug(C2p)gucaguUfuGfuucuuscsu
3335
AGAAGAACAAACUGACGCAGAGU
3596





AD-1069884.1
asgsaacaAfaCfUfGfacgcagaguuL96
3075
asAfscucu(G2p)cgucagUfuUfguucususc
3336
GAAGAACAAACUGACGCAGAGUA
3597





AD-565919.1
gsasacaaAfcUfGfAfcgcagaguauL96
3076
asUfsacuc(Tgn)gcgucaGfuUfuguucsusu
3337
AAGAACAAACUGACGCAGAGUAA
3598





AD-1069885.1
asascaaaCfuGfAfCfgcagaguaauL96
3077
asUfsuacu(C2p)ugcgucAfgUfuuguuscsu
3338
AGAACAAACUGACGCAGAGUAAG
3599





AD-565921.1
ascsaaacUfgAfCfGfcagaguaaguL96
3078
asCfsuuac(Tgn)cugcguCfaGfuuugususc
3339
GAACAAACUGACGCAGAGUAAGA
3600





AD-1069886.1
csasaacuGfaCfGfCfagaguaagauL96
3079
asUfscuua(C2p)ucugcgUfcAfguuugsusu
3340
AACAAACUGACGCAGAGUAAGAU
3601





AD-565923.1
asasacugAfcGfCfAfgaguaagauuL96
3080
asAfsucuu(Agn)cucugcGfuCfaguuusgsu
3341
ACAAACUGACGCAGAGUAAGAUC
3602





AD-565924.1
asascugaCfgCfAfGfaguaagaucuL96
3081
asGfsaucu(Tgn)acucugCfgUfcaguususg
3342
CAAACUGACGCAGAGUAAGAUCU
3603





AD-1069887.1
csusgacgCfaGfAfGfuaagaucuguL96
3082
asCfsagau(C2p)uuacucUfgCfgucagsusu
3343
AACUGACGCAGAGUAAGAUCUGG
3604





AD-565927.1
usgsacgcAfgAfGfUfaagaucugguL96
3083
asCfscaga(Tgn)cuuacuCfuGfcgucasgsu
3344
ACUGACGCAGAGUAAGAUCUGGG
3605





AD-565928.1
gsascgcaGfaGfUfAfagaucuggguL96
3084
asCfsccag(Agn)ucuuacUfcUfgcgucsasg
3345
CUGACGCAGAGUAAGAUCUGGGA
3606





AD-1069888.1
ascsgcagAfgUfAfAfgaucugggauL96
3085
asUfsccca(G2p)aucuuaCfuCfugcguscsa
3346
UGACGCAGAGUAAGAUCUGGGAC
3607





AD-566379.1
usgsagcaUfgUfCfGfgacaagaaauL96
3086
asUfsuucu(Tgn)guccgaCfaUfgcucascsa
3347
UGUGAGCAUGUCGGACAAGAAAG
3608





AD-566380.1
gsasgcauGfuCfGfGfacaagaaaguL96
3087
asCfsuuuc(Tgn)uguccgAfcAfugcucsasc
3348
GUGAGCAUGUCGGACAAGAAAGG
3609





AD-1069889.1
asgscaugUfcGfGfAfcaagaaagguL96
3088
asCfscuuu(C2p)uuguccGfaCfaugcuscsa
3349
UGAGCAUGUCGGACAAGAAAGGG
3610





AD-566382.1
gscsauguCfgGfAfCfaagaaaggguL96
3089
asCfsccuu(Tgn)cuugucCfgAfcaugcsusc
3350
GAGCAUGUCGGACAAGAAAGGGA
3611





AD-566383.2
csasugucGfgAfCfAfagaaagggauL96
3090
asUfscccu(Tgn)ucuuguCfcGfacaugscsu
3351
AGCAUGUCGGACAAGAAAGGGAU
3612





AD-566384.2
asusgucgGfaCfAfAfgaaagggauuL96
3091
asAfsuccc(Tgn)uucuugUfcCfgacausgsc
3352
GCAUGUCGGACAAGAAAGGGAUC
3613





AD-1069890.1
usgsucggAfcAfAfGfaaagggaucuL96
3092
asGfsaucc(C2p)uuucuuGfuCfcgacasusg
3353
CAUGUCGGACAAGAAAGGGAUCU
3614





AD-1069891.1
gsuscggaCfaAfGfAfaagggaucuuL96
3093
asAfsgauc(C2p)cuuucuUfgUfccgacsasu
3354
AUGUCGGACAAGAAAGGGAUCUG
3615





AD-1069892.1
uscsggacAfaGfAfAfagggaucuguL96
3094
asCfsagau(C2p)ccuuucUfuGfuccgascsa
3355
UGUCGGACAAGAAAGGGAUCUGU
3616





AD-566388.2
csgsgacaAfgAfAfAfgggaucuguuL96
3095
asAfscaga(Tgn)cccuuuCfuUfguccgsasc
3356
GUCGGACAAGAAAGGGAUCUGUG
3617





AD-566389.1
gsgsacaaGfaAfAfGfgg aucuguguL96
3096
asCfsacag(Agn)ucccuuUfcUfuguccsgsa
3357
UCGGACAAGAAAGGGAUCUGUGU
3618





AD-1069893.1
gsascaagAfaAfGfGfgaucuguguuL96
3097
asAfscaca(G2p)aucccuUfuCfuugucscsg
3358
CGGACAAGAAAGGGAUCUGUGUG
3619





AD-566391.1
ascsaagaAfaGfGfGfaucuguguguL96
3098
asCfsacac(Agn)gaucccUfuUfcuuguscsc
3359
GGACAAGAAAGGGAUCUGUGUGG
3620





AD-1069894.1
csasagaaAfgGfGfAfucugugugguL96
3099
asCfscaca(C2p)agauccCfuUfucuugsusc
3360
GACAAGAAAGGGAUCUGUGUGGC
3621





AD-566393.1
asasgaaaGfgGfAfUfcuguguggcuL96
3100
asGfsccac(Agn)cagaucCfcUfuucuusgsu
3361
ACAAGAAAGGGAUCUGUGUGGCA
3622





AD-566395.1
gsasaaggGfaUfCfUfguguggcaguL96
3101
asCfsugcc(Agn)cacagaUfcCfcuuucsusu
3362
AAGAAAGGGAUCUGUGUGGCAGA
3623





AD-1069896.1
asasagggAfuCfUfGfuguggcagauL96
3102
asUfscugc(C2p)acacagAfuCfccuuuscsu
3363
AGAAAGGGAUCUGUGUGGCAGAC
3624





AD-1069897.1
asasgggaUfcUfGfUfguggcagacuL96
3103
asGfsucug(C2p)cacacaGfaUfcccuususc
3364
GAAAGGGAUCUGUGUGGCAGACC
3625





AD-1069898.1
asgsggauCfuGfUfGfuggcagaccuL96
3104
asGfsgucu(G2p)ccacacAfgAfucccususu
3365
AAAGGGAUCUGUGUGGCAGACCC
3626





AD-1069899.1
gsgsgaucUfgUfGfUfggcagacccuL96
3105
asGfsgguc(Tgn)gccacaCfaGfaucccsusu
3366
AAGGGAUCUGUGUGGCAGACCCC
3627





AD-566475.1
gsasaaucCfgAfGfCfcguucucuauL96
3106
asUfsagag(Agn)acggcuCfgGfauuucscsa
3367
UGGAAAUCCGAGCCGUUCUCUAC
3628





AD-1069900.1
asasauccGfaGfCfCfguucucuacuL96
3107
asGfsuaga(G2p)aacggcUfcGfgauuuscsc
3368
GGAAAUCCGAGCCGUUCUCUACA
3629





AD-566477.1
asasuccgAfgCfCfGfuucucuacauL96
3108
asUfsguag(Agn)gaacggCfuCfggauususc
3369
GAAAUCCGAGCCGUUCUCUACAA
3630





AD-1069901.1
asusccgaGfcCfGfUfucucuacaauL96
3109
asUfsugua(G2p)agaacgGfcUfcggaususu
3370
AAAUCCGAGCCGUUCUCUACAAU
3631





AD-566483.1
asgsccguUfcUfCfUfacaauuaccuL96
3110
asGfsguaa(Tgn)uguagaGfaAfcggcuscsg
3371
CGAGCCGUUCUCUACAAUUACCG
3632





AD-566484.1
gscscguuCfuCfUfAfcaauuaccguL96
3111
asCfsggua(Agn)uuguagAfgAfacggcsusc
3372
GAGCCGUUCUCUACAAUUACCGG
3633





AD-566485.2
cscsguucUfcUfAfCfaauuaccgguL96
3112
asCfscggu(Agn)auuguaGfaGfaacggscsu
3373
AGCCGUUCUCUACAAUUACCGGC
3634





AD-566486.1
csgsuucuCfuAfCfAfauuaccggcuL96
3113
asGfsccgg(Tgn)aauuguAfgAfgaacgsgsc
3374
GCCGUUCUCUACAAUUACCGGCA
3635





AD-1069902.1
gsusucucUfaCfAfAfuuaccggcauL96
3114
asUfsgccg(G2p)uaauugUfaGfagaacsgsg
3375
CCGUUCUCUACAAUUACCGGCAG
3636





AD-1069903.1
ususcucuAfcAfAfUfuaccggcaguL96
3115
asCfsugcc(G2p)guaauuGfuAfgagaascsg
3376
CGUUCUCUACAAUUACCGGCAGA
3637





AD-1069904.1
uscsucuaCfaAfUfUfaccggcagauL96
3116
asUfscugc(C2p)gguaauUfgUfagagasasc
3377
GUUCUCUACAAUUACCGGCAGAA
3638





AD-1069905.1
gsgscugaCfcGfCfCfuacguggucuL96
3117
asGfsacca(C2p)guaggcGfgUfcagccsasg
3378
CUGGCUGACCGCCUACGUGGUCA
3639





AD-567054.1
gscsugacCfgCfCfUfacguggucauL96
3118
asUfsgacc(Agn)cguaggCfgGfucagcscsa
3379
UGGCUGACCGCCUACGUGGUCAA
3640





AD-1069906.1
csusgaccGfcCfUfAfcguggucaauL96
3119
asUfsugac(C2p)acguagGfcGfgucagscsc
3380
GGCUGACCGCCUACGUGGUCAAG
3641





AD-1069907.1
usgsaccgCfcUfAfCfguggucaaguL96
3120
asCfsuuga(C2p)cacguaGfgCfggucasgsc
3381
GCUGACCGCCUACGUGGUCAAGG
3642





AD-567057.1
gsasccgcCfuAfCfGfuggucaagguL96
3121
asCfscuug(Agn)ccacguAfgGfcggucsasg
3382
CUGACCGCCUACGUGGUCAAGGU
3643





AD-1069908.1
ascscgccUfaCfGfUfggucaagguuL96
3122
asAfsccuu(G2p)accacgUfaGfgcgguscsa
3383
UGACCGCCUACGUGGUCAAGGUC
3644





AD-567059.1
cscsgccuAfcGfUfGfgucaaggucuL96
3123
asGfsaccu(Tgn)gaccacGfuAfggcggsusc
3384
GACCGCCUACGUGGUCAAGGUCU
3645





AD-567060.1
csgsccuaCfgUfGfGfucaaggucuuL96
3124
asAfsgacc(Tgn)ugaccaCfgUfaggcgsgsu
3385
ACCGCCUACGUGGUCAAGGUCUU
3646





AD-1069909.1
gscscuacGfuGfGfUfcaaggucuuuL96
3125
asAfsagac(C2p)uugaccAfcGfuaggcsgsg
3386
CCGCCUACGUGGUCAAGGUCUUC
3647





AD-1069910.1
cscsuacgUfgGfUfCfaaggucuucuL96
3126
asGfsaaga(C2p)cuugacCfaCfguaggscsg
3387
CGCCUACGUGGUCAAGGUCUUCU
3648





AD-567063.4
csusacguGfgUfCfAfaggucuucuuL96
3127
asAfsgaag(Agn)ccuugaCfcAfcguagsgsc
3388
GCCUACGUGGUCAAGGUCUUCUC
3649





AD-1069911.1
usascgugGfuCfAfAfggucuucucuL96
3128
asGfsagaa(G2p)accuugAfcCfacguasgsg
3389
CCUACGUGGUCAAGGUCUUCUCU
3650





AD-567065.1
ascsguggUfcAfAfGfgucuucucuuL96
3129
asAfsgaga(Agn)gaccuuGfaCfcacgusasg
3390
CUACGUGGUCAAGGUCUUCUCUC
3651





AD-567066.4
csgsugguCfaAfGfGfucuucucucuL96
3130
asGfsagag(Agn)agaccuUfgAfccacgsusa
3391
UACGUGGUCAAGGUCUUCUCUCU
3652





AD-1069912.1
gsusggucAfaGfGfUfcuucucucuuL96
3131
asAfsgaga(G2p)aagaccUfuGfaccacsgsu
3392
ACGUGGUCAAGGUCUUCUCUCUG
3653





AD-567068.1
usgsgucaAfgGfUfCfuucucucuguL96
3132
asCfsagag(Agn)gaagacCfuUfgaccascsg
3393
CGUGGUCAAGGUCUUCUCUCUGG
3654





AD-1069913.1
gsgsucaaGfgUfCfUfucucucugguL96
3133
asCfscaga(G2p)agaagaCfcUfugaccsasc
3394
GUGGUCAAGGUCUUCUCUCUGGC
3655





AD-567070.1
gsuscaagGfuCfUfUfcucucuggcuL96
3134
asGfsccag(Agn)gagaagAfcCfuugacscsa
3395
UGGUCAAGGUCUUCUCUCUGGCU
3656





AD-1069914.1
uscsaaggUfcUfUfCfucucuggcuuL96
3135
asAfsgcca(G2p)agagaaGfaCfcuugascsc
3396
GGUCAAGGUCUUCUCUCUGGCUG
3657





AD-567072.1
csasagguCfuUfCfUfcucuggcuguL96
3136
asCfsagcc(Agn)gagagaAfgAfccuugsasc
3397
GUCAAGGUCUUCUCUCUGGCUGU
3658





AD-1069915.1
asasggucUfuCfUfCfucuggcuguuL96
3137
asAfscagc(C2p)agagagAfaGfaccuusgsa
3398
UCAAGGUCUUCUCUCUGGCUGUC
3659





AD-1069916.1
asgsgucuUfcUfCfUfcuggcugucuL96
3138
asGfsacag(C2p)cagagaGfaAfgaccususg
3399
CAAGGUCUUCUCUCUGGCUGUCA
3660





AD-1069917.1
gsgsucuuCfuCfUfCfuggcugucauL96
3139
asUfsgaca(G2p)ccagagAfgAfagaccsusu
3400
AAGGUCUUCUCUCUGGCUGUCAA
3661





AD-567076.1
gsuscuucUfcUfCfUfggcugucaauL96
3140
asUfsugac(Agn)gccagaGfaGfaagacscsu
3401
AGGUCUUCUCUCUGGCUGUCAAC
3662





AD-1069918.1
uscsuucuCfuCfUfGfgcugucaacuL96
3141
asGfsuuga(C2p)agccagAfgAfgaagascsc
3402
GGUCUUCUCUCUGGCUGUCAACC
3663





AD-567294.1
usasaagcAfgGfAfGfacuuccuuguL96
3142
asCfsaagg(Agn)agucucCfuGfcuuuasgsu
3403
ACUAAAGCAGGAGACUUCCUUGA
3664





AD-1069919.1
asasagcaGfgAfGfAfcuuccuugauL96
3143
asUfscaag(G2p)aagucuCfcUfgcuuusasg
3404
CUAAAGCAGGAGACUUCCUUGAA
3665





AD-1069920.1
asasgcagGfaGfAfCfuuccuugaauL96
3144
asUfsucaa(G2p)gaagucUfcCfugcuususa
3405
UAAAGCAGGAGACUUCCUUGAAG
3666





AD-567297.1
asgscaggAfgAfCfUfuccuugaaguL96
3145
asCfsuuca(Agn)ggaaguCfuCfcugcususu
3406
AAAGCAGGAGACUUCCUUGAAGC
3667





AD-567300.1
asgsgagaCfuUfCfCfuugaagccauL96
3146
asUfsggcu(Tgn)caaggaAfgUfcuccusgsc
3407
GCAGGAGACUUCCUUGAAGCCAA
3668





AD-567301.1
gsgsagacUfuCfCfUfugaagccaauL96
3147
asUfsuggc(Tgn)ucaaggAfaGfucuccsusg
3408
CAGGAGACUUCCUUGAAGCCAAC
3669





AD-1069922.1
gsasgacuUfcCfUfUfgaagccaacuL96
3148
asGfsuugg(C2p)uucaagGfaAfgucucscsu
3409
AGGAGACUUCCUUGAAGCCAACU
3670





AD-1069923.1
asgsacuuCfcUfUfGfaagccaacuuL96
3149
asAfsguug(G2p)cuucaaGfgAfagucuscsc
3410
GGAGACUUCCUUGAAGCCAACUA
3671





AD-1069924.1
gsascuucCfuUfGfAfagccaacuauL96
3150
asUfsaguu(G2p)gcuucaAfgGfaagucsusc
3411
GAGACUUCCUUGAAGCCAACUAC
3672





AD-567305.1
ascsuuccUfuGfAfAfgccaacuacuL96
3151
asGfsuagu(Tgn)ggcuucAfaGfgaaguscsu
3412
AGACUUCCUUGAAGCCAACUACA
3673





AD-567306.1
csusuccuUfgAfAfGfccaacuacauL96
3152
asUfsguag(Tgn)uggcuuCfaAfggaagsusc
3413
GACUUCCUUGAAGCCAACUACAU
3674





AD-567308.1
uscscuugAfaGfCfCfaacuacauguL96
3153
asCfsaugu(Agn)guuggcUfuCfaaggasasg
3414
CUUCCUUGAAGCCAACUACAUGA
3675





AD-567309.1
cscsuugaAfgCfCfAfacuacaugauL96
3154
asUfscaug(Tgn)aguuggCfuUfcaaggsasa
3415
UUCCUUGAAGCCAACUACAUGAA
3676





AD-1069925.1
csusugaaGfcCfAfAfcuacaugaauL96
3155
asUfsucau(G2p)uaguugGfcUfucaagsgsa
3416
UCCUUGAAGCCAACUACAUGAAC
3677





AD-567311.1
ususgaagCfcAfAfCfuacaugaacuL96
3156
asGfsuuca(Tgn)guaguuGfgCfuucaasgsg
3417
CCUUGAAGCCAACUACAUGAACC
3678





AD-567312.1
usgsaagcCfaAfCfUfacaugaaccuL96
3157
asGfsguuc(Agn)uguaguUfgGfcuucasasg
3418
CUUGAAGCCAACUACAUGAACCU
3679





AD-1069926.1
gsasagccAfaCfUfAfcaugaaccuuL96
3158
asAfsgguu(C2p)auguagUfuGfgcuucsasa
3419
UUGAAGCCAACUACAUGAACCUA
3680





AD-567314.2
asasgccaAfcUfAfCfaugaaccuauL96
3159
asUfsaggu(Tgn)cauguaGfuUfggcuuscsa
3420
UGAAGCCAACUACAUGAACCUAC
3681





AD-567315.6
asgsccaaCfuAfCfAfugaaccuacuL96
3160
asGfsuagg(Tgn)ucauguAfgUfuggcususc
3421
GAAGCCAACUACAUGAACCUACA
3682





AD-1069927.1
gscscaacUfaCfAfUfgaaccuacauL96
3161
asUfsguag(G2p)uucaugUfaGfuuggcsusu
3422
AAGCCAACUACAUGAACCUACAG
3683





AD-1069928.1
cscsaacuAfcAfUfGfaaccuacaguL96
3162
asCfsugua(G2p)guucauGfuAfguuggscsu
3423
AGCCAACUACAUGAACCUACAGA
3684





AD-567318.2
csasacuaCfaUfGfAfaccuacagauL96
3163
asUfscugu(Agn)gguucaUfgUfaguugsgsc
3424
GCCAACUACAUGAACCUACAGAG
3685





AD-567319.1
asascuacAfuGfAfAfccuacagaguL96
3164
asCfsucug(Tgn)agguucAfuGfuaguusgsg
3425
CCAACUACAUGAACCUACAGAGA
3686





AD-1069929.1
ascsuacaUfgAfAfCfcuacagagauL96
3165
asUfscucu(G2p)uagguuCfaUfguagususg
3426
CAACUACAUGAACCUACAGAGAU
3687





AD-567321.1
csusacauGfaAfCfCfuacagagauuL96
3166
asAfsucuc(Tgn)guagguUfcAfuguagsusu
3427
AACUACAUGAACCUACAGAGAUC
3688





AD-1069930.1
usascaugAfaCfCfUfacagagaucuL96
3167
asGfsaucu(C2p)uguaggUfuCfauguasgsu
3428
ACUACAUGAACCUACAGAGAUCC
3689





AD-567323.1
ascsaugaAfcCfUfAfcagagauccuL96
3168
asGfsgauc(Tgn)cuguagGfuUfcaugusasg
3429
CUACAUGAACCUACAGAGAUCCU
3690





AD-1069931.1
csasugaaCfcUfAfCfagagauccuuL96
3169
asAfsggau(C2p)ucuguaGfgUfucaugsusa
3430
UACAUGAACCUACAGAGAUCCUA
3691





AD-567325.1
asusgaacCfuAfCfAfgagauccuauL96
3170
asUfsagga(Tgn)cucuguAfgGfuucausgsu
3431
ACAUGAACCUACAGAGAUCCUAC
3692





AD-567326.1
usgsaaccUfaCfAfGfagauccuacuL96
3171
asGfsuagg(Agn)ucucugUfaGfguucasusg
3432
CAUGAACCUACAGAGAUCCUACA
3693





AD-1069932.1
gsasaccuAfcAfGfAfgauccuacauL96
3172
asUfsguag(G2p)aucucuGfuAfgguucsasu
3433
AUGAACCUACAGAGAUCCUACAC
3694





AD-1069933.1
asasccuaCfaGfAfGfauccuacacuL96
3173
asGfsugua(G2p)gaucucUfgUfagguuscsa
3434
UGAACCUACAGAGAUCCUACACU
3695





AD-567479.1
gsgscccuAfcUfGfCfagcuaaaaguL96
3174
asCfsuuuu(Agn)gcugcaGfuAfgggccsasa
3435
UUGGCCCUACUGCAGCUAAAAGA
3696





AD-567480.1
gscsccuaCfuGfCfAfgcuaaaagauL96
3175
asUfscuuu(Tgn)agcugcAfgUfagggcscsa
3436
UGGCCCUACUGCAGCUAAAAGAC
3697





AD-567481.1
cscscuacUfgCfAfGfcuaaaagacuL96
3176
asGfsucuu(Tgn)uagcugCfaGfuagggscsc
3437
GGCCCUACUGCAGCUAAAAGACU
3698





AD-567482.1
cscsuacuGfcAfGfCfuaaaagacuuL96
3177
asAfsgucu(Tgn)uuagcuGfcAfguaggsgsc
3438
GCCCUACUGCAGCUAAAAGACUU
3699





AD-1069934.1
usascugcAfgCfUfAfaaagacuuuuL96
3178
asAfsaagu(C2p)uuuuagCfuGfcaguasgsg
3439
CCUACUGCAGCUAAAAGACUUUG
3700





AD-567485.1
ascsugcaGfcUfAfAfaagacuuuguL96
3179
asCfsaaag(Tgn)cuuuuaGfcUfgcagusasg
3440
CUACUGCAGCUAAAAGACUUUGA
3701





AD-1069935.1
csusgcagCfuAfAfAfagacuuugauL96
3180
asUfscaaa(G2p)ucuuuuAfgCfugcagsusa
3441
UACUGCAGCUAAAAGACUUUGAC
3702





AD-567487.2
usgscagcUfaAfAfAfgacuuugacuL96
3181
asGfsucaa(Agn)gucuuuUfaGfcugcasgsu
3442
ACUGCAGCUAAAAGACUUUGACU
3703





AD-567488.1
gscsagcuAfaAfAfGfacuuugacuuL96
3182
asAfsguca(Agn)agucuuUfuAfgcugcsasg
3443
CUGCAGCUAAAAGACUUUGACUU
3704





AD-567489.1
csasgcuaAfaAfGfAfcuuugacuuuL96
3183
asAfsaguc(Agn)aagucuUfuUfagcugscsa
3444
UGCAGCUAAAAGACUUUGACUUU
3705





AD-1069936.1
asgscuaaAfaGfAfCfuuugacuuuuL96
3184
asAfsaagu(C2p)aaagucUfuUfuagcusgsc
3445
GCAGCUAAAAGACUUUGACUUUG
3706





AD-567491.1
gscsuaaaAfgAfCfUfuugacuuuguL96
3185
asCfsaaag(Tgn)caaaguCfuUfuuagcsusg
3446
CAGCUAAAAGACUUUGACUUUGU
3707





AD-1069937.1
gsusgccuCfcCfGfUfcgugcguuguL96
3186
asCfsaacg(C2p)acgacgGfgAfggcacsasa
3447
UUGUGCCUCCCGUCGUGCGUUGG
3708





AD-1069938.1
usgsccucCfcGfUfCfgugcguugguL96
3187
asCfscaac(G2p)cacgacGfgGfaggcascsa
3448
UGUGCCUCCCGUCGUGCGUUGGC
3709





AD-1069939.1
gscscuccCfgUfCfGfugcguuggcuL96
3188
asGfsccaa(C2p)gcacgaCfgGfgaggcsasc
3449
GUGCCUCCCGUCGUGCGUUGGCU
3710





AD-567513.1
cscsucccGfuCfGfUfgcguuggcuuL96
3189
asAfsgcca(Agn)cgcacgAfcGfggaggscsa
3450
UGCCUCCCGUCGUGCGUUGGCUC
3711





AD-567514.1
csuscccgUfcGfUfGfcguuggcucuL96
3190
asGfsagcc(Agn)acgcacGfaCfgggagsgsc
3451
GCCUCCCGUCGUGCGUUGGCUCA
3712





AD-1069940.1
uscsccguCfgUfGfCfguuggcucauL96
3191
asUfsgagc(C2p)aacgcaCfgAfcgggasgsg
3452
CCUCCCGUCGUGCGUUGGCUCAA
3713





AD-1069941.1
cscscgucGfuGfCfGfuuggcucaauL96
3192
asUfsugag(C2p)caacgcAfcGfacgggsasg
3453
CUCCCGUCGUGCGUUGGCUCAAU
3714





AD-1069942.1
cscsgucgUfgCfGfUfuggcucaauuL96
3193
asAfsuuga(G2p)ccaacgCfaCfgacggsgsa
3454
UCCCGUCGUGCGUUGGCUCAAUG
3715





AD-567518.1
csgsucguGfcGfUfUfggcucaauguL96
3194
asCfsauug(Agn)gccaacGfcAfcgacgsgsg
3455
CCCGUCGUGCGUUGGCUCAAUGA
3716





AD-1069943.1
gsuscgugCfgUfUfGfgcucaaugauL96
3195
asUfscauu(G2p)agccaaCfgCfacgacsgsg
3456
CCGUCGUGCGUUGGCUCAAUGAA
3717





AD-567521.4
csgsugcgUfuGfGfCfucaaugaacuL96
3196
asGfsuuca(Tgn)ugagccAfaCfgcacgsasc
3457
GUCGUGCGUUGGCUCAAUGAACA
3718





AD-1069944.1
usgscguuGfgCfUfCfaaugaacaguL96
3197
asCfsuguu(C2p)auugagCfcAfacgcascsg
3458
CGUGCGUUGGCUCAAUGAACAGA
3719





AD-567524.1
gscsguugGfcUfCfAfaugaacagauL96
3198
asUfscugu(Tgn)cauugaGfcCfaacgcsasc
3459
GUGCGUUGGCUCAAUGAACAGAG
3720





AD-567525.1
csgsuuggCfuCfAfAfugaacagaguL96
3199
asCfsucug(Tgn)ucauugAfgCfcaacgscsa
3460
UGCGUUGGCUCAAUGAACAGAGA
3721





AD-1069945.1
gsusuggcUfcAfAfUfgaacagagauL96
3200
asUfscucu(G2p)uucauuGfaGfccaacsgsc
3461
GCGUUGGCUCAAUGAACAGAGAU
3722





AD-567527.1
ususggcuCfaAfUfGfaacagagauuL96
3201
asAfsucuc(Tgn)guucauUfgAfgccaascsg
3462
CGUUGGCUCAAUGAACAGAGAUA
3723





AD-1069946.1
usgsgcucAfaUfGfAfacagagauauL96
3202
asUfsaucu(C2p)uguucaUfuGfagccasasc
3463
GUUGGCUCAAUGAACAGAGAUAC
3724





AD-567529.1
gsgscucaAfuGfAfAfcagagauacuL96
3203
asGfsuauc(Tgn)cuguucAfuUfgagccsasa
3464
UUGGCUCAAUGAACAGAGAUACU
3725





AD-1069947.1
gscsucaaUfgAfAfCfagagauacuuL96
3204
asAfsguau(C2p)ucuguuCfaUfugagcscsa
3465
UGGCUCAAUGAACAGAGAUACUA
3726





AD-567531.1
csuscaauGfaAfCfAfgagauacuauL96
3205
asUfsagua(Tgn)cucuguUfcAfuugagscsc
3466
GGCUCAAUGAACAGAGAUACUAC
3727





AD-567532.1
uscsaaugAfaCfAfGfagauacuacuL96
3206
asGfsuagu(Agn)ucucugUfuCfauugasgsc
3467
GCUCAAUGAACAGAGAUACUACG
3728





AD-567533.1
csasaugaAfcAfGfAfgauacuacguL96
3207
asCfsguag(Tgn)aucucuGfuUfcauugsasg
3468
CUCAAUGAACAGAGAUACUACGG
3729





AD-1069948.1
asasugaaCfaGfAfGfauacuacgguL96
3208
asCfscgua(G2p)uaucucUfgUfucauusgsa
3469
UCAAUGAACAGAGAUACUACGGU
3730





AD-567535.1
asusgaacAfgAfGfAfuacuacgguuL96
3209
asAfsccgu(Agn)guaucuCfuGfuucaususg
3470
CAAUGAACAGAGAUACUACGGUG
3731





AD-568149.1
gsasgcagUfcAfAfGfgucuacgccuL96
3210
asGfsgcgu(Agn)gaccuuGfaCfugcucscsa
3471
UGGAGCAGUCAAGGUCUACGCCU
3732





AD-568150.1
asgscaguCfaAfGfGfucuacgccuuL96
3211
asAfsggcg(Tgn)agaccuUfgAfcugcuscsc
3472
GGAGCAGUCAAGGUCUACGCCUA
3733





AD-1069949.1
gscsagucAfaGfGfUfcuacgccuauL96
3212
asUfsaggc(G2p)uagaccUfuGfacugcsusc
3473
GAGCAGUCAAGGUCUACGCCUAU
3734





AD-1069950.1
csasgucaAfgGfUfCfuacgccuauuL96
3213
asAfsuagg(C2p)guagacCfuUfgacugscsu
3474
AGCAGUCAAGGUCUACGCCUAUU
3735





AD-1069951.1
asgsucaaGfgUfCfUfacgccuauuuL96
3214
asAfsauag(G2p)cguagaCfcUfugacusgsc
3475
GCAGUCAAGGUCUACGCCUAUUA
3736





AD-1069952.1
gsuscaagGfuCfUfAfcgccuauuauL96
3215
asUfsaaua(G2p)gcguagAfcCfuugacsusg
3476
CAGUCAAGGUCUACGCCUAUUAC
3737





AD-568155.1
uscsaaggUfcUfAfCfgccuauuacuL96
3216
asGfsuaau(Agn)ggcguaGfaCfcuugascsu
3477
AGUCAAGGUCUACGCCUAUUACA
3738





AD-568159.1
gsgsucuaCfgCfCfUfauuacaaccuL96
3217
asGfsguug(Tgn)aauaggCfgUfagaccsusu
3478
AAGGUCUACGCCUAUUACAACCU
3739





AD-1069953.1
gsuscuacGfcCfUfAfuuacaaccuuL96
3218
asAfsgguu(G2p)uaauagGfcGfuagacscsu
3479
AGGUCUACGCCUAUUACAACCUG
3740





AD-568161.2
uscsuacgCfcUfAfUfuacaaccuguL96
3219
asCfsaggu(Tgn)guaauaGfgCfguagascsc
3480
GGUCUACGCCUAUUACAACCUGG
3741





AD-568162.1
csusacgcCfuAfUfUfacaaccugguL96
3220
asCfscagg(Tgn)uguaauAfgGfcguagsasc
3481
GUCUACGCCUAUUACAACCUGGA
3742





AD-1069954.1
usascgccUfaUfUfAfcaaccuggauL96
3221
asUfsccag(G2p)uuguaaUfaGfgcguasgsa
3482
UCUACGCCUAUUACAACCUGGAG
3743





AD-1069955.1
ascsgccuAfuUfAfCfaaccuggaguL96
3222
asCfsucca(G2p)guuguaAfuAfggcgusasg
3483
CUACGCCUAUUACAACCUGGAGG
3744





AD-568165.1
csgsccuaUfuAfCfAfaccuggagguL96
3223
asCfscucc(Agn)gguuguAfaUfaggcgsusa
3484
UACGCCUAUUACAACCUGGAGGA
3745





AD-1069956.1
gscsugagGfaGfAfAfuugcuucauuL96
3224
asAfsugaa(G2p)caauucUfcCfucagcsasc
3485
GUGCUGAGGAGAAUUGCUUCAUA
3746





AD-568337.1
gscscaggAfgUfGfGfacuauguguuL96
3225
asAfscaca(Tgn)aguccaCfuCfcuggcsusc
3486
GAGCCAGGAGUGGACUAUGUGUA
3747





AD-568338.1
cscsaggaGfuGfGfAfcuauguguauL96
3226
asUfsacac(Agn)uaguccAfcUfccuggscsu
3487
AGCCAGGAGUGGACUAUGUGUAC
3748





AD-1069957.1
csasggagUfgGfAfCfuauguguacuL96
3227
asGfsuaca(C2p)auagucCfaCfuccugsgsc
3488
GCCAGGAGUGGACUAUGUGUACA
3749





AD-568340.1
asgsgaguGfgAfCfUfauguguacauL96
3228
asUfsguac(Agn)cauaguCfcAfcuccusgsg
3489
CCAGGAGUGGACUAUGUGUACAA
3750





AD-1069958.1
gsgsagugGfaCfUfAfuguguacaauL96
3229
asUfsugua(C2p)acauagUfcCfacuccsusg
3490
CAGGAGUGGACUAUGUGUACAAG
3751





AD-568342.1
gsasguggAfcUfAfUfguguacaaguL96
3230
asCfsuugu(Agn)cacauaGfuCfcacucscsu
3491
AGGAGUGGACUAUGUGUACAAGA
3752





AD-568343.4
asgsuggaCfuAfUfGfuguacaagauL96
3231
asUfscuug(Tgn)acacauAfgUfccacuscsc
3492
GGAGUGGACUAUGUGUACAAGAC
3753





AD-1069959.1
gsusggacUfaUfGfUfguacaagacuL96
3232
asGfsucuu(G2p)uacacaUfaGfuccacsusc
3493
GAGUGGACUAUGUGUACAAGACC
3754





AD-568345.2
usgsgacuAfuGfUfGfuacaagaccuL96
3233
asGfsgucu(Tgn)guacacAfuAfguccascsu
3494
AGUGGACUAUGUGUACAAGACCC
3755





AD-568348.1
ascsuaugUfgUfAfCfaagacccgauL96
3234
asUfscggg(Tgn)cuuguaCfaCfauaguscsc
3495
GGACUAUGUGUACAAGACCCGAC
3756





AD-1069961.1
csusauguGfuAfCfAfagacccgacuL96
3235
asGfsucgg(G2p)ucuuguAfcAfcauagsusc
3496
GACUAUGUGUACAAGACCCGACU
3757
















TABLE 24







C3 Single Dose Screens in PCH cells (% C3 mRNA Remaining)




















FU*

FU*

FU*

TX#

TX#

TX#



Duplex
500 nM
SD
100 nM
SD
10 nM
SD
10 nM
SD
1 nM
SD
0.1 nM
SD






















AD-570137.1
60.5
19.0
73.6
24.1
71.8
43.2
1.8
0.4
9.8
4.1
31.8
4.2


AD-570138.1
71.5
7.9
127.1
40.6
67.4
49.1
6.1
1.2
15.4
4.5
50.4
6.0


AD-570139.1
84.2
40.6
111.1
42.9
90.7
14.7
2.1
0.8
28.6
6.1
67.7
21.1


AD-570140.1
166.3
20.6
91.3
22.5
79.0
42.1
1.6
0.6
18.1
4.4
87.8
22.5


AD-570141.1
118.0
10.4
130.1
48.1
66.0
26.4
2.1
0.2
29.3
13.9
111.3
30.1


AD-570142.1
112.0
14.8
115.6
49.1
74.7
27.4
2.9
0.5
31.8
9.4
99.5
25.8


AD-570143.1
69.9
32.7
85.0
28.8
73.5
7.2
2.1
1.3
6.2
2.4
34.8
11.0


AD-570144.1
49.6
14.8
123.7
29.1
103.3
78.1
5.2
1.0
39.9
11.8
139.4
34.9


AD-570145.1
65.9
25.8
102.9
50.0
106.6
54.4
2.9
1.3
30.7
16.0
58.1
5.3


AD-570146.1
132.3
36.8
129.1
30.7
90.9
19.4
1.2
0.6
11.3
3.2
22.1
5.4


AD-570147.1
82.8
18.5
118.5
6.4
92.5
20.4
38.4
3.6
75.5
22.9
66.1
19.5


AD-570148.1
54.2
13.6
139.0
8.0
95.3
36.3
1.6
0.4
19.0
8.1
107.8
53.3


AD-570149.1
70.4
13.4
157.7
38.0
112.1
20.8
15.0
6.3
64.0
5.2
143.8
35.2


AD-570150.1
125.5
29.6
110.1
4.3
146.3
35.4
10.7
2.0
80.2
12.2
118.3
18.1


AD-570151.1
86.9
15.4
141.7
12.1
99.0
15.0
5.4
0.5
61.6
11.1
128.3
29.8


AD-570152.1
61.1
15.8
110.4
52.4
88.7
22.4
3.7
2.0
30.4
5.0
120.7
33.3


AD-570153.1
44.1
1.1
106.0
62.0
74.7
45.3
24.4
3.3
75.8
41.7
78.3
11.0


AD-570154.1
58.9
10.0
168.1
5.8
87.3
15.6
1.6
0.5
21.5
15.3
50.2
6.3


AD-570155.1
93.9
13.0
112.4
13.3
76.5
31.1
2.7
0.5
33.5
7.5
98.1
33.0


AD-570156.2
88.6
16.8
123.8
12.0
73.0
19.0
2.6
1.5
22.4
7.4
58.4
14.5


AD-570158.1
81.0
21.1
93.8
18.5
116.6
37.7
1.1
0.2
20.6
9.3
73.6
46.3


AD-570159.1
79.9
13.8
93.8
8.7
100.4
33.7
23.4
4.9
77.5
6.6
165.8
22.2


AD-570160.1
48.5
25.0
92.4
44.8
99.9
37.6
10.0
4.5
100.1
8.0
182.5
57.7


AD-570161.1
37.2
2.2
95.6
72.1
76.6
52.9
8.6
2.5
51.4
44.7
56.1
5.4


AD-570611.1
56.2
8.4
N/A
N/A
81.5
11.2
30.8
8.0
52.3
27.7
79.7
8.0


AD-570612.1
81.2
20.4
153.7
37.5
125.4
73.3
96.6
19.3
126.3
48.5
111.1
27.8


AD-570613.1
113.4
19.6
142.9
15.7
116.8
41.2
136.3
29.1
112.9
29.0
145.2
73.4


AD-570614.1
60.7
14.4
145.3
35.2
148.5
15.5
98.4
27.0
110.5
6.1
181.3
61.8


AD-570615.1
67.6
13.1
124.5
25.9
136.2
29.4
36.0
28.6
149.9
105.5
153.5
53.7


AD-570616.1
56.0
32.5
101.8
47.2
105.6
19.4
14.5
7.1
69.7
14.8
112.2
24.5


AD-570617.1
52.2
25.3
121.4
64.7
59.8
15.0
79.2
29.0
54.9
22.2
84.3
33.5


AD-570618.1
26.6
6.0
126.6
41.5
73.8
19.2
3.6
0.3
42.0
32.6
59.6
10.1


AD-570619.1
41.3
7.8
108.4
18.4
82.0
5.1
3.6
2.8
36.7
27.2
62.0
20.1


AD-570620.3
67.8
16.3
142.3
32.1
99.0
23.2
8.6
0.8
81.7
45.6
78.5
8.1


AD-570621.2
39.1
3.8
123.1
19.3
116.1
31.4
61.3
19.4
86.5
9.3
144.6
46.4


AD-570622.2
25.5
8.2
131.5
29.2
151.2
51.7
5.7
0.9
78.3
39.9
88.9
8.6


AD-570623.4
51.0
9.1
99.7
24.2
111.6
53.1
6.1
3.3
81.9
41.1
143.9
27.5


AD-570624.2
80.6
20.6
100.6
46.7
97.1
31.4
43.2
13.0
111.9
54.8
170.3
41.9


AD-570625.2
44.4
13.1
96.8
57.1
59.2
25.8
14.0
5.4
49.0
25.6
73.9
17.4


AD-570626.1
71.6
20.1
108.2
24.0
94.2
72.7
6.7
4.5
58.7
26.7
55.3
4.1


AD-570627.2
56.7
17.0
98.3
6.5
99.1
12.6
18.0
7.4
90.7
40.7
67.0
14.0


AD-570628.1
79.4
8.8
134.5
11.0
118.7
61.1
18.9
3.1
82.4
36.4
91.8
21.1


AD-570629.1
68.2
22.0
128.7
29.6
114.8
7.1
68.2
26.9
108.8
40.0
129.3
35.5


AD-570630.1
37.5
11.2
107.3
5.6
125.3
38.8
67.7
13.5
121.8
52.2
127.2
22.5


AD-1069837.1
28.4
3.9
81.3
9.7
165.3
36.1
129.7
47.9
104.3
28.4
113.7
11.9


AD-570707.1
81.8
43.9
80.3
46.8
48.2
16.5
1.0
0.2
8.1
2.7
27.7
2.4


AD-570708.1
65.2
18.9
141.0
18.7
66.1
40.2
9.3
1.3
53.2
30.9
47.7
19.1


AD-570709.1
34.8
14.1
128.6
32.0
72.7
15.4
23.4
4.1
88.9
50.0
40.9
8.7


AD-570710.1
73.8
10.7
157.4
19.7
108.1
16.8
31.8
9.8
113.8
52.8
52.9
10.9


AD-570715.1
65.6
7.5
119.5
31.9
109.4
12.7
3.3
1.4
22.4
5.6
47.2
7.2


AD-570716.1
72.6
27.0
113.2
18.7
111.8
26.5
3.5
2.3
41.0
7.8
48.8
16.7


AD-570717.2
69.6
12.6
89.4
28.8
119.1
32.5
16.2
2.8
99.0
20.6
71.2
29.7


AD-570718.1
29.5
10.9
82.9
36.8
132.7
18.8
3.4
0.9
78.7
30.1
27.0
10.6


AD-570719.1
65.9
43.7
66.0
33.7
60.2
26.8
1.8
1.0
9.7
3.9
21.4
3.9


AD-570720.1
62.6
37.2
132.0
26.0
75.9
20.9
33.1
4.9
67.2
45.6
66.8
14.8


AD-570721.1
38.2
22.5
111.5
20.3
91.5
20.4
8.0
4.2
63.5
23.1
57.1
18.2


AD-571285.1
39.5
15.1
120.7
25.8
90.5
13.6
115.2
36.2
125.4
60.7
94.3
17.6


AD-571286.1
62.7
2.4
126.1
13.5
91.6
32.2
26.4
3.1
79.4
43.2
92.5
49.0


AD-571287.1
64.9
9.9
114.4
9.1
105.4
16.4
171.9
56.1
88.1
39.8
94.4
18.9


AD-571288.1
37.9
12.1
86.4
22.2
112.9
41.2
153.0
27.6
81.0
11.7
106.9
29.8


AD-571289.1
41.8
10.2
82.0
37.5
117.3
45.1
34.6
9.5
83.8
17.9
99.4
5.5


AD-571290.1
65.8
30.0
98.5
40.2
54.1
22.5
74.8
29.1
74.7
50.1
79.5
12.5


AD-571291.1
114.1
14.4
142.5
31.4
104.0
24.3
76.6
14.0
98.7
36.8
64.6
10.3


AD-571292.1
70.6
13.9
93.3
4.8
123.4
34.8
1.4
0.6
28.9
10.1
62.2
17.7


AD-571293.1
70.7
28.1
96.6
21.1
114.4
21.1
1.6
0.7
36.5
20.7
73.7
8.1


AD-571294.1
63.6
8.8
126.3
50.3
94.7
18.7
6.7
2.8
69.2
37.0
84.9
9.6


AD-571295.1
31.5
8.7
79.5
20.0
125.2
45.9
1.9
1.0
25.9
15.6
52.1
13.9


AD-571296.1
68.1
29.7
66.6
30.0
87.3
24.8
1.1
0.6
14.3
2.2
36.5
6.9


AD-571297.1
62.1
15.9
83.5
25.5
55.2
9.2
3.1
1.4
37.3
16.2
65.6
29.0


AD-571298.6
82.7
18.1
125.1
20.1
94.5
25.7
2.6
0.4
19.9
9.9
36.8
6.8


AD-571299.1
94.6
19.6
73.2
14.4
79.3
33.8
0.9
0.6
13.7
3.4
20.4
3.4


AD-571300.1
64.3
8.3
92.0
12.2
97.8
43.9
2.1
1.2
29.8
19.5
47.1
7.2


AD-571301.1
81.4
15.7
92.2
14.8
77.6
17.4
19.1
5.5
104.5
35.4
85.8
16.8


AD-571302.1
80.2
23.4
69.5
10.4
76.3
35.1
3.4
0.3
43.2
14.9
57.4
13.1


AD-571303.1
67.2
25.9
72.7
42.9
62.2
6.4
3.2
0.8
51.3
6.9
65.1
27.4


AD-571304.1
18.6
4.4
78.4
29.7
56.3
21.8
3.0
0.7
39.7
10.4
62.4
10.1


AD-571305.1
74.6
30.3
103.8
6.7
82.1
23.9
3.2
2.2
16.8
4.7
38.6
4.1


AD-571306.1
42.0
11.8
90.3
31.1
78.5
35.7
4.6
1.7
22.4
12.4
56.7
13.3


AD-571307.1
56.0
20.3
61.1
13.5
67.1
9.0
1.1
0.3
13.1
5.3
24.9
6.6


AD-571308.1
64.3
21.8
80.2
15.9
104.8
32.7
3.1
1.0
25.9
9.6
50.6
5.6


AD-571309.1
51.6
9.0
96.8
41.1
113.7
18.4
4.8
1.9
39.8
25.3
67.5
8.0


AD-571526.1
43.3
8.6
88.2
29.2
137.1
29.5
10.8
1.5
67.2
27.8
57.5
7.1


AD-571527.1
36.8
6.5
60.2
14.1
72.1
27.8
2.1
0.5
16.3
7.9
42.0
8.0


AD-571528.1
64.0
9.5
50.3
11.4
63.8
19.3
1.4
0.4
3.7
1.4
15.7
5.7


AD-571529.1
60.6
15.6
88.0
20.9
97.1
36.5
6.3
1.4
46.0
20.7
49.0
15.3


AD-571530.1
92.8
16.5
98.1
27.6
76.9
47.0
18.7
8.9
57.0
18.6
56.4
10.8


AD-571531.1
92.5
11.1
87.3
2.1
58.0
26.4
5.2
1.8
31.3
16.7
54.2
7.0


AD-571532.1
71.6
27.8
70.9
9.6
61.7
16.6
2.3
0.6
8.5
4.0
28.6
2.6


AD-571533.1
41.5
12.5
46.4
7.6
65.6
28.5
1.3
0.4
4.0
5.1
10.3
2.3


AD-571534.1
46.7
5.6
79.7
20.5
66.7
25.1
2.5
0.8
15.1
2.4
42.8
13.6






#Transfection (TX)



*Free Uptake (FU)













TABLE 25







C3 Single Dose Screens in PCH cells (% C3 mRNA Remaining)




















FU*

FU*

FU*

TX#

TX#

TX#



Duplex
500 nM
SD
100 nM
SD
10 nM
SD
10 nM
SD
1 nM
SD
0.1 nM
SD






















AD-568955.1
63.2
9.9
61.5
17.9
97.0
34.4
1.8
0.8
15.3
3.7
28.0
4.4


AD-568956.1
65.4
2.0
93.0
23.7
123.9
26.8
3.1
0.3
59.5
22.9
53.7
11.9


AD-568957.1
55.5
7.3
78.3
15.0
88.8
6.3
3.2
1.5
25.3
12.7
37.8
15.6


AD-568958.1
96.8
14.2
85.5
20.8
N/A
N/A
4.8
1.5
82.1
23.7
72.7
13.4


AD-568959.1
87.4
12.1
84.2
42.0
126.8
22.7
3.8
0.4
80.4
26.4
64.3
20.6


AD-568960.1
100.2
5.5
70.3
3.1
117.6
11.3
14.9
5.6
141.7
22.9
79.0
23.6


AD-568961.1
92.0
7.2
91.9
23.2
114.3
34.7
5.6
0.8
88.2
36.9
59.1
19.9


AD-568962.1
83.1
20.6
91.5
15.2
98.0
14.8
4.4
1.4
36.7
9.1
57.0
34.0


AD-568963.2
53.6
11.2
73.8
36.3
107.6
28.6
1.6
0.8
20.3
7.0
35.5
9.5


AD-568964.1
72.4
3.6
89.2
24.9
106.7
1.6
8.5
4.6
85.6
20.1
52.1
16.5


AD-568965.1
42.7
1.7
76.6
22.0
66.5
11.0
1.2
0.1
56.3
50.2
24.8
8.0


AD-568966.1
58.2
7.2
79.8
13.4
83.0
16.5
2.0
1.7
82.4
34.2
42.5
25.0


AD-568967.1
96.5
3.9
87.6
11.0
88.9
13.3
4.8
2.4
99.8
28.1
40.5
17.8


AD-568968.1
88.0
8.0
94.6
18.8
85.3
10.1
3.1
1.0
129.0
60.6
54.4
15.0


AD-568969.1
53.2
5.7
73.9
23.4
88.9
26.4
2.7
1.0
22.0
8.6
37.0
12.5


AD-568970.1
85.9
12.8
80.1
22.5
111.1
6.6
4.0
0.7
60.9
20.3
54.2
14.8


AD-568971.1
58.4
12.8
73.4
31.0
105.1
27.8
2.8
3.1
10.5
2.3
34.4
11.8


AD-568972.1
48.7
8.3
70.6
14.8
99.9
26.1
1.9
1.6
9.6
2.2
22.4
4.3


AD-568973.1
59.5
3.7
72.9
4.4
72.7
3.8
1.9
1.3
18.4
5.7
58.9
38.2


AD-568974.1
67.4
2.4
84.0
9.9
78.8
6.2
1.7
0.5
23.3
14.2
44.7
20.3


AD-568975.1
42.8
7.8
54.5
7.0
65.1
12.3
1.1
0.3
9.6
2.5
8.6
1.9


AD-568977.1
67.2
11.2
78.7
26.8
92.3
24.4
2.3
0.7
13.3
3.7
19.8
5.8


AD-568979.1
92.6
9.5
135.6
46.5
91.1
4.8
5.1
2.6
92.7
9.9
40.3
17.1


AD-1069834.1
99.1
17.1
39.6
10.7
90.5
41.0
1.9
0.4
37.4
13.6
41.3
31.1


AD-1069835.1
94.1
11.7
74.3
9.5
94.5
15.5
3.7
0.8
44.7
4.0
50.0
3.3


AD-1069836.1
78.1
8.3
84.3
11.7
92.9
15.6
3.0
0.7
45.0
16.4
75.6
16.6


AD-569154.1
115.3
28.0
108.7
17.5
101.9
19.2
36.3
5.0
120.2
19.0
65.5
12.1


AD-569155.1
93.0
7.7
82.7
8.9
85.3
8.4
6.4
1.6
92.2
24.5
49.3
14.1


AD-569156.1
63.8
6.5
79.8
8.3
96.3
30.2
3.1
1.3
22.2
4.1
38.8
17.1


AD-569157.1
58.5
13.6
75.3
11.6
80.7
11.8
1.5
0.2
8.8
2.4
15.5
1.8


AD-569158.1
67.8
3.1
78.5
42.2
83.5
23.5
1.8
0.3
11.5
5.7
27.4
12.6


AD-569159.1
50.1
9.4
66.5
14.7
89.9
21.2
1.5
0.3
9.2
1.8
24.6
11.5


AD-569160.1
61.7
8.6
86.8
16.6
89.4
9.9
2.0
1.6
7.7
1.9
19.6
1.9


AD-569161.1
64.9
11.6
79.6
8.9
90.4
3.6
2.2
1.3
13.6
2.7
38.6
14.7


AD-569162.1
105.3
6.1
117.7
9.9
96.8
17.9
41.7
6.4
73.4
23.9
35.6
5.8


AD-569163.1
59.6
5.1
88.4
19.4
74.3
17.0
1.5
0.3
13.3
3.4
36.7
24.0


AD-569166.1
114.2
32.3
100.1
16.5
84.0
11.8
6.6
0.7
59.2
11.5
53.7
16.0


AD-569167.1
106.8
21.9
85.8
30.9
98.8
13.3
7.3
1.1
98.6
15.9
46.7
26.0


AD-569168.1
78.5
8.3
51.7
17.4
103.3
26.6
7.8
0.4
66.8
34.2
25.6
3.2


AD-569169.1
90.2
8.5
84.5
17.3
122.3
16.7
2.6
0.1
41.0
25.0
19.5
10.9


AD-569170.1
101.9
9.1
98.3
16.2
112.4
14.8
45.9
11.1
57.9
8.3
57.4
23.2


AD-569171.1
117.3
5.8
106.7
18.1
107.1
11.1
62.2
4.9
93.8
43.6
56.8
12.8


AD-569172.1
94.1
7.1
107.3
17.3
91.5
13.4
34.2
9.8
80.8
15.1
59.2
27.5


AD-569173.1
94.3
9.1
102.1
17.9
98.5
14.7
20.5
2.4
79.7
12.3
50.7
31.9


AD-569174.1
90.4
11.8
107.0
13.9
93.3
10.2
52.9
6.8
121.6
30.3
63.7
22.8


AD-569175.1
93.6
7.1
68.2
8.1
86.6
20.7
9.8
3.4
34.1
7.0
41.5
22.6


AD-569262.1
14.8
6.0
38.5
6.4
68.7
11.9
0.8
0.3
5.5
1.8
6.7
6.2


AD-569263.1
24.6
3.1
47.2
1.8
82.3
18.0
1.3
0.5
14.2
14.4
9.4
7.0


AD-569264.1
28.1
4.9
47.5
5.8
72.4
2.7
1.3
0.1
5.5
1.3
8.8
1.9


AD-569265.1
31.5
0.9
48.5
5.4
65.2
9.3
1.7
1.6
4.5
1.0
8.2
5.8


AD-569266.1
27.1
4.2
51.3
8.2
61.9
13.2
0.9
0.4
10.4
10.5
15.0
8.7


AD-569267.1
31.0
2.1
47.8
4.5
68.9
11.0
2.7
2.8
5.6
0.5
15.6
10.3


AD-569268.1
13.2
2.1
31.6
6.6
57.7
22.4
1.0
0.5
4.0
0.5
1.1
0.4


AD-569269.1
17.1
2.8
30.0
15.2
46.8
10.4
1.2
0.5
3.2
1.3
4.7
4.4


AD-569270.1
31.3
4.8
42.9
8.3
80.9
20.1
0.9
0.1
5.0
2.7
7.6
1.2


AD-569271.1
36.2
19.9
59.3
5.9
76.5
12.6
1.0
0.2
9.0
5.3
9.1
4.6


AD-569273.1
72.4
18.1
106.7
26.2
113.3
22.6
2.5
0.3
31.5
10.4
25.7
11.2


AD-569274.1
51.7
2.4
76.1
14.4
82.5
13.3
1.6
0.3
10.7
2.3
31.9
8.2


AD-569275.1
108.1
16.0
105.2
6.0
102.9
9.8
28.4
7.8
82.5
23.1
52.3
37.7


AD-569276.1
83.6
10.6
86.9
9.6
112.3
14.5
4.4
1.2
48.4
12.9
38.1
10.6


AD-569277.1
69.0
6.0
85.1
16.2
102.3
40.6
2.4
1.0
19.5
4.3
49.3
47.4


AD-569278.1
102.5
19.7
62.3
1.8
80.2
19.9
24.7
3.7
51.6
10.5
48.3
33.1


AD-569279.1
113.3
28.3
105.6
7.2
108.8
24.7
78.8
7.4
73.3
20.9
47.6
15.9


AD-569280.1
103.2
12.0
121.9
22.0
96.4
12.3
62.7
4.6
74.6
7.1
56.5
11.8


AD-569281.1
98.3
8.6
109.2
15.7
96.2
16.0
84.4
26.7
87.4
33.8
48.6
17.7


AD-569282.1
106.1
5.7
92.0
1.5
98.5
11.3
113.7
20.7
86.2
28.3
30.2
2.6


AD-569506.1
85.5
3.6
114.8
23.0
93.7
11.4
5.9
3.6
43.0
18.6
42.6
8.8


AD-569507.1
76.8
6.5
105.7
35.9
87.6
20.7
2.1
0.6
19.9
4.8
28.4
9.1


AD-569508.1
73.6
5.9
75.0
32.4
67.4
19.5
3.4
1.7
18.4
8.3
29.8
7.2


AD-569509.1
79.2
15.0
82.9
8.6
94.8
13.2
3.5
1.8
25.8
5.2
46.5
10.2


AD-569510.1
45.4
7.4
71.9
2.9
81.1
11.8
2.7
2.0
8.7
2.4
18.0
9.9


AD-569511.1
34.1
5.6
57.9
14.2
68.6
5.4
1.5
0.5
7.0
1.1
17.7
10.2


AD-569512.1
70.5
7.9
111.5
7.4
76.4
16.4
5.0
3.8
28.3
9.7
41.2
14.0


AD-569513.1
80.0
16.1
107.8
19.0
91.3
20.3
2.5
1.2
19.0
2.1
26.7
6.5


AD-569514.1
28.4
2.5
62.9
23.9
70.8
11.9
1.2
0.3
4.6
0.6
11.3
4.7


AD-569515.1
58.7
6.4
61.8
22.7
55.2
12.0
3.2
0.5
19.8
5.7
20.3
12.6


AD-569516.1
71.0
5.5
111.5
19.0
91.1
1.4
3.8
0.7
37.2
7.1
51.3
26.2


AD-569517.1
95.4
12.9
78.1
11.7
96.7
11.1
2.2
0.7
13.2
4.5
27.1
11.8


AD-569518.1
97.2
6.6
97.2
9.3
116.1
12.5
12.6
2.4
61.1
33.7
76.4
34.0


AD-569519.1
87.1
8.9
103.7
29.5
80.8
10.0
6.5
0.6
58.8
20.8
40.1
16.2


AD-569520.1
75.5
4.0
100.6
15.9
99.5
26.2
2.5
0.3
32.0
5.6
33.2
1.1


AD-569565.1
67.9
9.9
79.4
11.1
97.4
6.2
2.1
0.4
13.8
1.1
21.7
6.1


AD-569567.1
61.8
9.3
83.3
25.9
84.9
10.5
1.7
0.6
11.6
2.3
18.1
4.8


AD-570126.1
107.5
16.9
63.4
12.9
98.8
30.6
32.2
16.0
50.1
8.7
34.6
6.2


AD-570127.1
52.2
1.4
69.2
15.8
69.8
10.8
3.9
1.7
6.1
2.1
19.5
1.8


AD-570128.1
104.2
17.0
78.4
4.4
92.9
27.3
6.8
1.4
38.1
12.8
44.3
20.2


AD-570129.1
113.3
18.9
71.3
15.9
96.7
15.0
23.1
8.1
50.8
18.9
36.5
7.8


AD-570131.1
75.6
14.5
81.3
14.8
101.2
16.5
2.7
1.1
15.0
4.7
35.5
10.0


AD-570135.1
69.5
9.8
64.6
22.3
78.9
7.7
1.6
0.7
12.1
2.2
20.1
3.1


AD-570136.1
52.8
8.4
66.2
16.4
73.4
4.9
1.3
0.3
6.5
0.9
9.0
2.8






#Transfection (TX)



*Free Uptake (FU)













TABLE 26







C3 Single Dose Screens in PCH cells (% C3 mRNA Remaining)




















FU*

FU*

FU*

TX#

TX#

TX#



Duplex
500 nM
SD
100 nM
SD
10 nM
SD
10 nM
SD
1 nM
SD
0.1 nM
SD






















AD-571535.1
79.2
8.9
83.4
10.0
87.7
29.6
75.7
29.2
N/A
N/A
93.9
19.1


AD-571536.1
50.2
6.5
45.0
7.5
102.8
25.6
93.4
16.8
54.5
29.7
51.2
12.9


AD-571537.1
46.4
4.9
42.1
6.8
93.3
14.1
96.6
21.5
85.3
7.5
44.9
5.7


AD-571538.1
80.6
7.4
74.1
8.0
173.4
51.1
108.2
24.0
73.1
84.9
153.3
19.0


AD-571540.1
68.4
12.7
64.3
13.5
132.7
35.0
150.0
51.9
54.2
26.0
100.2
7.2


AD-571541.1
108.6
25.7
80.9
5.1
165.1
38.3
147.7
16.4
84.8
12.8
147.7
47.6


AD-571542.1
49.1
6.4
48.0
4.8
144.1
46.2
193.0
127.6
36.5
14.3
35.0
10.0


AD-571543.1
55.7
10.2
52.3
7.0
127.5
43.5
126.3
46.9
47.3
39.6
69.1
24.3


AD-571544.1
74.3
14.1
49.0
8.7
82.0
36.5
76.2
16.3
9.3
8.5
78.5
21.1


AD-571545.1
82.3
4.9
77.4
7.5
104.5
24.5
96.7
8.3
96.3
7.1
105.6
15.8


AD-571546.1
64.4
25.3
53.0
3.0
72.0
11.8
100.3
14.5
88.4
9.5
46.3
5.5


AD-571547.1
36.9
7.7
39.6
3.8
72.4
18.0
131.2
32.1
124.1
75.5
20.5
4.3


AD-571548.1
56.8
17.3
64.6
8.3
80.8
16.9
125.7
6.9
131.3
154.6
40.6
7.3


AD-571549.1
114.2
26.2
99.2
19.1
110.3
33.9
119.0
16.3
211.2
74.0
127.6
34.8


AD-571550.1
69.9
4.8
68.2
16.3
92.9
23.8
142.0
32.5
43.0
8.2
90.4
22.0


AD-571551.1
89.0
32.8
71.7
7.7
130.0
30.4
150.4
30.2
49.6
45.5
148.0
5.7


AD-571552.1
82.3
18.6
75.0
13.8
109.0
32.1
82.3
14.4
29.8
18.3
68.9
17.2


AD-571553.1
41.5
4.2
55.0
1.2
72.9
18.3
96.2
9.5
42.3
13.3
18.2
6.4


AD-571554.1
74.0
12.6
64.1
7.6
98.7
9.4
111.4
7.3
92.2
42.0
58.1
12.7


AD-571555.1
86.5
16.5
96.8
12.5
108.8
14.1
107.4
6.5
67.0
38.0
135.9
16.3


AD-571556.1
75.4
19.7
88.5
4.4
106.0
18.6
119.6
14.9
58.4
35.2
111.8
9.0


AD-571557.1
59.8
12.1
66.8
4.9
80.8
4.1
148.8
44.8
68.7
20.8
19.0
3.5


AD-571558.1
73.3
24.1
62.1
6.0
107.7
33.0
111.8
11.6
125.1
33.4
59.4
12.2


AD-571559.1
91.2
14.0
80.5
21.3
104.5
22.1
87.6
13.4
28.2
11.4
44.8
4.7


AD-571560.1
48.0
12.4
66.6
4.4
86.5
11.9
122.6
4.1
39.6
30.5
27.0
8.6


AD-571711.1
102.2
11.3
112.9
16.7
112.1
9.8
108.7
6.3
125.7
93.2
117.4
21.6


AD-571712.1
96.4
11.0
89.8
9.8
105.2
11.4
97.5
21.2
87.5
46.2
113.7
28.1


AD-571713.1
55.7
7.5
69.0
5.1
104.6
28.1
128.9
18.2
109.0
25.8
89.4
14.2


AD-571714.1
68.3
9.5
68.2
8.8
94.1
5.6
123.6
24.1
84.7
57.5
105.8
14.3


AD-571716.1
96.0
11.1
60.0
11.5
87.8
14.7
101.3
21.8
55.5
12.2
113.2
8.7


AD-571717.1
74.9
15.5
62.8
6.9
103.5
32.1
95.4
22.2
43.8
19.7
33.6
5.2


AD-571718.1
27.4
4.1
45.5
6.3
71.8
15.2
204.5
82.6
84.7
133.8
19.8
4.5


AD-571719.2
31.9
3.6
57.1
6.2
98.9
29.4
171.1
37.0
109.2
130.1
32.1
3.8


AD-571720.1
67.0
4.8
77.0
11.4
95.1
11.1
193.3
61.4
19.7
8.0
40.3
11.4


AD-571721.1
35.8
6.7
48.2
5.6
79.0
10.6
130.7
34.1
32.2
17.1
21.2
9.6


AD-571722.1
22.2
4.7
35.1
5.5
84.9
15.3
150.9
72.0
125.1
103.5
24.1
13.8


AD-571723.1
49.0
12.1
64.9
8.0
97.4
16.1
234.7
140.7
124.5
47.1
67.2
16.2


AD-571742.1
106.4
2.6
87.8
17.6
113.3
31.5
168.2
79.2
42.5
13.9
209.9
24.0


AD-571743.1
89.9
17.3
104.8
27.6
112.3
54.1
81.0
20.6
64.9
17.6
40.0
11.7


AD-571744.1
88.1
18.2
106.7
9.7
133.8
56.9
121.1
19.3
61.0
20.5
46.8
15.5


AD-571745.1
66.1
15.6
96.2
14.4
99.6
5.0
100.0
12.8
N/A
N/A
64.5
14.6


AD-571746.1
114.5
25.5
120.1
14.6
136.5
31.4
91.8
3.7
83.6
48.4
82.3
7.2


AD-571747.1
82.6
11.3
89.6
8.3
109.5
13.0
76.7
37.8
40.0
23.5
75.0
19.0


AD-571748.1
30.2
5.5
57.5
7.4
87.7
11.5
108.5
19.3
48.0
14.2
31.3
6.0


AD-571749.1
29.6
3.2
55.3
5.8
79.1
8.4
106.2
8.9
22.3
18.9
28.5
3.3


AD-571750.1
107.4
11.5
95.7
21.2
115.5
52.4
86.1
22.2
N/A
N/A
39.6
10.7


AD-571751.1
81.4
12.8
101.6
13.1
101.4
11.0
102.5
17.3
25.4
24.7
44.0
4.7


AD-571753.2
36.4
9.3
52.6
6.3
85.8
7.0
102.8
18.4
85.5
34.7
31.5
7.0


AD-571755.1
81.5
21.0
91.3
8.0
111.7
18.1
103.3
15.1
43.2
30.7
73.7
8.6


AD-571756.1
98.2
14.2
106.6
37.5
116.4
17.5
101.3
13.4
126.1
55.2
78.7
24.4


AD-571757.1
64.3
5.7
75.7
10.9
105.9
17.9
115.5
29.6
39.9
25.2
63.0
11.8


AD-571758.1
90.3
11.1
93.6
11.9
114.7
44.8
108.9
23.8
34.5
18.4
109.3
16.6


AD-571759.1
49.6
9.8
42.9
5.8
69.8
7.8
89.0
14.9
67.4
23.9
52.3
20.2


AD-571760.1
63.3
4.7
72.5
7.0
91.4
37.3
82.2
22.6
33.7
15.1
18.5
8.6


AD-571761.1
54.1
3.5
70.9
8.7
82.3
14.6
126.3
20.6
15.2
4.0
25.6
4.9


AD-571762.1
37.2
3.9
63.6
7.8
74.1
6.0
116.4
18.3
98.1
34.9
28.3
4.3


AD-571763.1
33.8
8.0
50.1
6.5
78.7
8.3
121.9
21.4
80.6
62.4
24.4
7.6


AD-571764.1
62.0
20.4
71.3
3.2
105.0
36.9
117.6
8.9
67.5
36.9
30.3
5.8


AD-571765.2
84.7
10.7
92.0
14.3
110.0
7.1
122.2
6.2
146.0
113.7
97.8
21.9


AD-571766.2
65.4
11.3
73.5
22.0
101.6
13.3
116.5
12.8
71.3
81.4
78.4
22.0


AD-571767.2
80.6
19.5
58.1
5.9
91.0
20.8
97.1
17.0
143.9
83.0
88.8
7.8


AD-572383.1
69.2
18.0
79.1
2.6
80.9
5.6
102.5
11.1
109.1
34.7
53.3
11.5


AD-572384.1
78.1
11.3
97.5
10.8
121.5
8.4
107.2
6.5
115.5
3.4
64.8
15.5


AD-572385.1
79.2
9.2
94.8
12.5
92.7
10.4
104.3
14.6
79.5
27.8
64.5
5.6


AD-572386.1
41.7
3.6
66.6
4.8
92.1
27.8
99.9
22.9
68.5
1.7
35.4
14.3


AD-572387.4
86.4
3.0
70.1
8.1
77.8
10.0
80.3
10.7
66.2
82.6
119.9
17.8


AD-572391.1
90.7
19.3
91.9
10.5
125.7
28.0
86.0
29.4
44.2
19.6
113.0
11.7


AD-572392.1
66.1
13.5
72.3
8.9
88.7
7.6
134.5
36.2
N/A
N/A
46.4
8.8


AD-572393.2
99.8
13.6
97.1
20.3
100.0
19.3
116.0
19.1
152.6
108.7
56.7
8.0


AD-572394.1
102.9
8.9
111.1
22.1
108.6
22.1
125.6
14.9
48.1
26.4
61.8
17.7


AD-572395.1
109.6
18.9
102.9
11.9
115.5
19.1
118.0
18.8
47.0
21.9
82.3
14.6


AD-572396.1
98.1
14.9
104.2
7.7
118.3
27.5
166.3
106.9
23.7
10.1
82.7
32.0


AD-572397.1
109.3
5.7
80.3
8.7
123.0
28.6
108.7
10.6
51.6
22.6
125.3
27.3


AD-572495.1
25.9
4.5
28.9
4.6
97.7
48.0
87.7
39.7
39.4
17.6
10.0
3.9


AD-572569.1
117.7
31.0
100.9
13.4
110.9
26.1
124.2
16.5
N/A
N/A
85.4
24.5


AD-572570.1
43.8
6.6
58.0
6.6
95.9
24.3
100.3
10.5
34.6
9.2
37.7
6.1


AD-572571.1
60.3
8.7
74.0
15.7
98.9
28.9
116.1
17.2
119.1
100.7
42.0
5.8


AD-572572.1
81.3
15.5
83.3
11.3
96.8
22.9
95.5
3.1
76.9
37.2
36.2
10.7


AD-572573.1
70.2
22.3
72.2
23.1
66.0
17.5
127.1
29.9
315.6
73.6
26.9
4.7


AD-572574.1
93.8
13.6
90.6
15.0
129.2
56.9
100.7
9.3
10.8
13.7
86.0
20.5


AD-572575.1
66.5
17.4
64.7
14.9
105.6
30.1
88.4
9.8
34.1
7.8
68.3
20.7


AD-572576.1
88.0
5.2
103.4
33.5
100.7
41.7
94.6
65.2
70.8
25.4
112.6
46.5


AD-572577.1
118.6
27.9
111.9
17.0
176.5
84.7
140.6
28.6
36.2
12.9
114.4
15.3


AD-572580.1
90.9
61.5
97.7
16.0
127.3
36.4
123.8
16.6
N/A
N/A
121.6
46.9


AD-572581.1
77.3
11.5
80.4
14.9
143.2
51.4
109.6
21.3
150.2
107.7
87.8
19.7






#Transfection (TX)



*Free Uptake (FU)













TABLE 27







C3 Single Dose Screens in PCH cells (% C3 mRNA Remaining)






















FU*
ST
FU*
ST
FU*
ST
TX#
ST
TX#
ST
TX#
ST
TX#
ST


Duplex
500 nM
DEV
100 nM
DEV
10 nM
DEV
50 nM
DEV
10 nM
DEV
1 nM
DEV
0.1 nM
DEV
























AD-564723.1
51.6
21.2
52.2
24.7
82.7
58.9
19.8
3.8
13.3
6.8
55.8
24.0
145.8
88.9


AD-564724.1
64.3
13.1
102.9
30.8
106.6
31.7
4.7
1.6
3.8
0.8
37.8
14.9
101.9
13.1


AD-1069838.1
110.5
19.4
94.8
13.3
139.4
65.9
36.0
6.5
16.8
5.9
87.5
3.5
106.5
9.7


AD-564726.1
140.5
42.4
129.4
77.4
127.2
10.8
14.9
1.8
13.6
3.9
84.2
6.7
75.9
12.4


AD-564727.3
120.7
43.2
149.1
107.3
158.0
46.3
8.1
1.2
7.1
2.4
65.8
20.3
79.0
14.1


AD-1069839.1
180.1
78.3
132.6
50.8
184.9
59.0
4.9
0.5
3.3
0.9
58.6
20.4
80.6
19.7


AD-1069840.1
122.7
26.8
164.0
78.4
181.7
52.7
23.8
6.3
13.6
6.7
65.0
20.5
77.4
17.2


AD-564730.3
64.5
14.5
151.1
55.1
214.4
86.6
0.7
0.2
1.0
0.4
7.5
4.5
32.0
12.6


AD-1069841.1
42.9
16.9
69.8
8.1
88.1
15.0
13.3
2.6
11.0
3.3
51.6
7.1
106.5
40.2


AD-564732.1
78.1
14.9
95.4
37.1
62.7
17.3
65.1
30.7
20.7
4.4
66.3
15.5
97.0
11.7


AD-1069842.1
63.4
21.5
91.6
20.6
81.5
16.9
0.7
0.1
0.4
0.1
1.5
0.2
44.1
9.6


AD-564734.1
49.7
12.5
97.1
38.6
91.8
31.4
1.1
0.1
0.5
0.1
1.2
0.4
21.5
5.8


AD-1069843.1
82.1
28.8
127.2
41.0
107.0
27.5
34.9
0.7
20.3
8.2
85.7
3.7
73.9
3.3


AD-564736.1
87.0
16.4
129.9
37.8
148.5
23.6
11.5
2.8
6.0
1.0
39.5
3.0
77.0
14.0


AD-1069844.1
97.3
70.9
156.6
63.5
143.4
46.3
1.1
0.1
0.6
0.1
2.1
0.6
36.8
5.5


AD-564738.1
79.8
49.6
189.2
57.4
212.1
52.0
1.1
0.2
0.8
0.2
8.9
2.9
70.8
60.8


AD-564739.2
62.4
27.8
67.3
11.7
67.1
20.4
3.5
0.6
1.9
0.2
3.2
0.9
77.4
13.2


AD-1069845.1
70.0
20.9
64.7
26.4
64.1
7.2
21.2
3.8
5.8
0.8
7.3
4.0
51.0
18.4


AD-564741.1
85.4
11.3
89.6
43.6
86.3
8.4
0.9
0.1
0.8
0.1
1.8
0.7
23.8
4.6


AD-1069846.1
70.5
6.6
85.3
12.3
92.7
16.2
0.9
0.2
1.6
2.0
1.3
0.4
16.7
3.5


AD-1069847.1
84.0
44.2
111.8
39.6
144.2
32.3
1.2
0.1
0.7
0.1
1.8
0.4
25.7
7.4


AD-564745.3
71.6
22.1
149.7
16.0
156.5
39.8
1.2
0.3
0.6
0.1
4.5
1.2
44.0
50.1


AD-564747.1
77.1
23.0
124.9
26.7
206.1
63.9
3.1
1.3
4.0
4.1
45.7
7.9
89.9
90.8


AD-1069850.1
116.0
12.9
117.3
51.4
88.8
18.0
77.9
32.4
65.4
15.5
82.9
7.4
116.3
11.0


AD-1069851.1
175.9
87.7
110.6
28.8
90.1
14.4
46.4
8.7
41.5
2.1
84.6
10.7
91.5
6.8


AD-1069852.1
83.9
29.4
142.7
54.9
114.9
23.9
6.1
1.4
6.3
1.5
58.8
7.1
96.5
11.4


AD-1069853.1
65.5
34.3
212.8
66.4
154.0
24.8
1.0
0.2
0.6
0.1
4.5
1.1
99.0
73.4


AD-564925.1
67.1
31.9
182.9
26.2
233.5
65.7
1.6
0.2
1.2
0.2
8.9
2.6
18.6
20.2


AD-1069854.1
55.6
27.1
57.9
35.7
53.7
38.2
1.7
0.2
1.6
0.5
3.7
0.8
53.5
21.8


AD-1069855.1
90.1
28.9
50.4
25.9
43.8
20.2
1.7
0.6
0.8
0.4
2.4
1.2
48.6
9.8


AD-1069856.1
119.0
43.7
63.1
15.4
56.3
8.3
3.9
0.7
2.3
1.2
4.9
0.2
83.1
13.8


AD-564929.1
133.2
31.0
94.5
48.8
68.6
12.6
54.0
5.6
45.9
13.3
60.8
5.7
95.0
15.0


AD-564930.1
137.8
39.7
112.8
11.8
80.8
10.0
94.4
46.1
43.2
9.3
72.4
7.3
105.3
19.5


AD-1069857.1
121.1
54.4
95.7
11.2
105.7
19.4
1.5
0.5
1.0
0.2
10.2
0.7
83.6
14.4


AD-564934.1
80.9
25.7
201.1
71.7
125.8
11.0
131.0
76.5
84.2
30.5
89.7
9.5
118.2
8.5


AD-1069858.1
110.6
55.7
138.5
63.5
166.6
26.4
30.6
5.0
44.7
21.2
102.6
13.1
81.0
21.4


AD-564936.1
109.6
55.7
53.8
16.1
47.5
8.5
34.0
9.7
72.7
35.1
67.1
18.7
95.8
17.7


AD-564937.1
89.3
54.3
78.9
23.9
44.1
9.4
23.4
6.2
24.8
6.1
29.4
10.4
87.0
50.7


AD-564938.1
114.2
28.9
119.7
94.1
77.4
15.3
116.8
48.7
117.9
12.2
88.9
11.7
105.9
12.7


AD-1069859.1
97.0
22.5
88.1
39.6
106.7
55.0
31.3
5.8
36.9
7.4
70.7
2.9
99.2
18.2


AD-564941.1
138.1
75.5
126.7
72.8
145.6
50.3
195.8
40.5
140.9
3.1
93.0
18.7
109.2
13.3


AD-1069860.1
143.9
45.8
146.3
62.2
141.7
35.7
128.5
73.3
118.5
12.8
110.3
13.5
112.8
36.5


AD-564943.1
107.4
92.7
50.2
20.1
47.1
14.3
15.1
1.0
19.9
9.7
21.4
7.6
85.5
7.1


AD-1069861.1
64.0
39.2
53.1
17.6
54.1
14.5
0.6
0.1
0.5
0.3
0.4
0.0
3.6
1.1


AD-565031.1
60.7
8.6
105.7
48.3
46.9
10.6
0.8
0.1
0.5
0.1
1.0
0.1
5.3
0.4


AD-565032.1
75.1
14.1
82.6
21.7
81.2
9.9
0.9
0.2
0.7
0.0
0.6
0.1
4.7
0.8


AD-1069862.1
59.8
15.1
93.2
15.2
84.4
9.4
0.8
0.1
0.9
0.1
0.9
0.1
7.6
0.9


AD-565034.1
38.2
10.1
65.7
22.4
76.9
7.8
0.7
0.1
0.6
0.2
0.8
0.2
5.2
0.6


AD-565035.1
30.8
5.7
88.6
16.3
96.5
23.9
0.7
0.1
0.7
0.3
0.6
0.1
3.2
0.6


AD-1069863.1
52.4
25.9
123.5
69.8
186.4
61.0
0.8
0.1
0.5
0.1
0.8
0.1
4.0
1.0


AD-565037.1
59.2
54.4
43.7
13.3
43.0
14.5
0.6
0.2
0.4
0.1
0.6
0.1
3.7
0.5


AD-565038.1
153.3
95.6
77.3
27.8
44.4
28.3
0.8
0.3
1.0
0.4
0.7
0.1
16.7
3.5


AD-1069864.1
78.0
26.5
64.1
38.5
44.9
8.1
2.5
0.3
1.2
0.2
1.6
0.3
34.8
14.1


AD-565041.1
147.5
44.2
89.8
25.9
67.7
20.2
7.5
2.2
10.5
2.5
29.8
9.7
80.3
10.7


AD-565042.1
119.3
51.9
106.0
40.0
81.9
31.9
26.3
7.5
31.8
6.1
54.7
9.5
93.3
20.9


AD-565043.1
118.8
19.8
96.4
24.0
131.5
60.2
78.8
26.5
92.7
13.0
102.3
6.5
106.2
15.4


AD-565044.1
138.1
65.0
120.5
55.1
94.7
29.2
140.6
35.1
106.6
7.4
113.1
16.4
119.9
37.6


AD-1069865.1
87.9
34.7
113.0
52.7
131.1
51.4
12.3
2.4
5.2
1.0
13.0
3.3
64.8
12.8


AD-1069866.1
100.3
23.2
51.0
11.4
69.9
25.1
95.0
58.7
150.1
16.7
105.2
7.5
100.4
27.0


AD-565047.1
89.4
30.3
75.5
13.4
92.8
29.8
146.5
50.0
165.3
20.7
96.5
10.3
119.5
19.8


AD-1069867.1
126.4
38.6
110.7
9.7
171.0
14.9
160.8
60.1
169.2
14.9
98.3
17.6
113.7
14.8


AD-565049.1
111.7
35.4
105.5
27.0
111.3
37.3
138.2
50.8
171.1
6.9
108.4
3.8
118.0
20.3


AD-565050.1
96.4
61.5
125.6
39.4
116.9
30.8
180.9
55.9
175.6
27.4
119.2
11.4
118.1
22.6


AD-565274.1
137.1
70.3
152.5
59.8
109.4
23.5
69.4
11.7
127.5
16.0
119.2
8.1
99.4
30.3


AD-565275.1
75.7
54.5
120.0
28.2
130.7
33.6
3.5
1.2
5.4
0.4
57.3
16.0
65.7
16.7


AD-1069868.1
105.4
79.2
77.0
29.7
50.2
19.7
3.5
1.2
2.5
0.1
12.0
3.7
103.7
9.2


AD-1069869.1
89.0
37.8
75.5
25.4
65.4
11.9
14.4
2.9
19.2
6.0
53.6
16.1
106.2
24.4


AD-565278.2
94.1
35.2
67.6
13.4
68.7
19.4
1.9
0.4
2.4
0.8
11.6
2.5
94.1
23.0


AD-1069870.1
66.1
37.2
61.6
1.6
70.4
9.1
0.8
0.2
0.8
0.4
1.8
0.4
28.6
4.3


AD-565280.1
71.8
21.2
88.2
40.0
165.3
112.7
56.9
25.4
54.1
15.8
80.2
7.7
114.3
18.0


AD-565281.3
49.5
22.2
79.9
24.8
79.9
16.7
5.9
2.0
3.6
0.5
9.9
2.7
74.0
8.1


AD-1069871.1
52.4
11.2
83.9
36.5
108.4
22.2
2.1
0.3
1.1
0.3
2.4
0.3
18.3
1.0


AD-565283.1
37.3
16.7
131.6
39.1
173.4
121.6
1.2
0.2
1.8
0.2
7.5
2.2
70.8
30.9


AD-1069872.1
80.4
25.9
84.2
44.3
71.8
42.0
1.8
0.4
1.5
0.5
7.9
3.9
81.9
29.0


AD-1069873.1
57.7
31.1
82.4
10.0
59.2
13.7
1.4
0.2
1.8
0.6
5.1
1.3
81.8
30.3


AD-565286.1
91.7
38.5
114.8
56.6
66.1
16.9
84.5
24.1
111.0
25.3
113.5
16.4
118.5
13.2


AD-565287.1
57.7
10.6
115.5
44.9
73.9
23.0
29.7
12.3
23.1
9.7
73.5
11.3
131.4
12.8


AD-1069874.1
49.9
10.3
72.7
13.1
87.0
36.6
32.5
6.6
71.2
64.3
109.3
56.2
160.1
64.6


AD-1069875.1
103.4
31.7
117.0
37.5
97.3
26.8
75.8
21.8
128.7
94.9
78.3
14.0
114.8
12.1


AD-565335.1
52.9
16.8
100.5
16.6
200.3
91.6
2.5
0.5
2.5
0.4
14.7
4.2
38.3
6.7


AD-1069876.1
46.4
18.4
46.7
25.1
70.2
21.6
29.2
12.9
74.9
13.1
93.8
8.9
35.5
12.4


AD-565895.1
77.5
56.1
69.7
22.4
93.6
44.3
1.7
0.7
1.7
0.1
5.5
0.6
62.9
14.8


AD-1069877.1
55.6
21.0
90.8
20.6
113.8
38.3
111.5
61.6
142.5
28.7
112.1
9.0
111.5
14.2


AD-565897.1
91.6
15.2
80.3
9.7
78.3
29.1
111.4
74.4
150.6
13.8
110.0
6.7
79.7
10.0


AD-565899.1
51.9
7.7
73.4
12.8
90.3
10.4
35.9
19.7
34.0
12.8
80.6
16.3
105.7
19.9


AD-565903.1
71.9
20.4
80.6
21.1
102.8
42.6
10.3
2.9
7.9
3.2
14.4
6.6
59.0
13.8


AD-565904.3
77.0
8.0
62.9
29.1
90.2
18.5
2.3
0.7
3.9
2.7
2.6
0.9
21.9
3.6






#Transfection (TX)



*Free Uptake (FU)













TABLE 28







C3 Single Dose Screens in PCH cells (% C3 mRNA Remaining)






















FU*

FU*

FU*

TX#

TX#

TX#

TX#



Duplex ID
500 nM
STDEV
100 nM
STDEV
10 nM
STDEV
50 nM
STDEV
10 nM
STDEV
1 nM
STDEV
0.1 nM
STDEV
























AD-1069878.1
51.3
5.7
62.2
36.8
214.9
37.4
4.2
0.6
1.8
0.3
3.8
2.0
254.9
63.1


AD-565906.1
59.0
2.5
60.6
38.4
202.5
44.2
120.6
10.1
103.4
30.2
95.3
14.1
242.6
53.0


AD-565907.1
62.6
10.5
39.9
18.9
232.0
103.1
0.7
0.3
1.3
0.3
9.6
2.0
278.0
123.5


AD-1069879.1
82.6
12.0
43.4
16.7
165.9
70.0
7.1
2.3
7.9
1.0
30.3
4.5
198.8
83.9


AD-565909.1
93.3
15.5
49.4
23.0
245.8
78.8
4.2
0.1
8.5
1.3
47.3
12.2
264.8
112.0


AD-565910.1
96.8
10.9
70.3
38.6
154.2
70.6
0.7
0.2
2.9
2.1
25.1
4.9
184.8
84.5


AD-565911.1
82.9
6.1
54.1
33.7
207.4
25.3
0.5
0.1
0.9
0.1
3.4
1.4
N/A
N/A


AD-1069880.1
98.1
26.1
64.0
32.2
N/A
N/A
11.6
6.0
36.3
3.5
101.2
16.7
N/A
N/A


AD-565913.1
67.5
17.9
32.8
4.3
141.2
69.8
2.9
0.4
4.4
1.1
71.0
25.0
169.2
83.6


AD-1069881.1
57.3
7.4
51.1
25.9
125.4
61.1
0.4
0.1
0.7
0.1
3.9
1.3
150.2
73.2


AD-565915.1
72.8
4.4
83.2
45.4
170.7
81.6
121.8
3.6
103.0
36.0
104.3
3.5
204.5
97.8


AD-1069882.1
65.1
10.5
58.2
42.0
151.4
60.5
0.4
0.1
1.0
0.3
8.4
1.3
181.3
72.5


AD-1069883.1
89.1
6.7
72.3
41.9
146.3
29.9
27.3
5.8
73.3
12.7
105.3
5.4
175.3
35.9


AD-1069884.1
104.0
10.5
44.0
15.7
198.2
32.0
51.0
10.7
72.9
23.9
112.5
7.6
237.5
38.3


AD-565919.1
126.6
29.1
34.6
12.2
225.4
54.6
81.9
9.2
126.6
14.3
112.5
7.9
275.8
54.7


AD-1069885.1
92.2
13.8
55.4
43.3
214.4
120.2
49.6
17.2
51.4
18.8
70.5
13.8
209.7
77.3


AD-565921.1
82.7
10.0
83.9
91.9
96.4
39.7
75.2
4.7
55.9
23.1
105.6
14.3
115.5
47.6


AD-1069886.1
70.1
13.8
89.6
100.6
117.8
49.1
1.1
0.5
2.4
0.6
30.9
8.4
141.1
58.8


AD-565923.1
85.8
3.8
59.6
26.2
134.9
18.9
93.9
6.2
90.2
21.5
91.9
2.4
161.7
22.6


AD-565924.1
84.2
12.1
80.5
31.1
151.3
72.1
1.2
0.3
1.9
0.2
12.7
1.1
181.2
86.4


AD-1069887.1
93.0
6.7
89.5
48.8
145.5
29.4
1.5
0.7
1.9
0.1
42.4
8.8
174.3
35.2


AD-565927.1
99.1
17.4
48.8
9.0
155.6
22.0
120.4
17.1
124.3
43.6
121.4
16.2
186.5
26.3


AD-565928.1
81.9
8.4
35.2
7.3
N/A
N/A
49.1
3.5
109.2
33.7
112.6
9.1
N/A
N/A


AD-1069888.1
74.5
15.4
44.3
21.0
101.9
48.2
2.8
0.6
10.6
5.8
71.0
5.5
122.1
57.8


AD-566379.1
83.0
7.7
109.1
77.5
100.6
44.5
102.3
27.4
104.5
49.0
85.6
6.6
120.5
53.3


AD-566380.1
91.9
8.7
128.7
35.6
122.5
10.5
109.5
6.1
124.9
7.0
113.4
12.1
146.8
12.6


AD-1069889.1
102.2
10.9
126.8
41.6
147.3
28.4
109.2
12.4
126.8
18.2
104.8
13.8
176.5
34.0


AD-566382.1
97.8
24.0
45.4
4.4
151.2
15.0
112.7
4.7
130.0
12.0
109.0
9.1
181.2
18.0


AD-566383.2
105.2
18.5
133.5
93.9
136.9
39.0
115.4
13.0
129.5
11.3
112.4
9.3
164.0
46.7


AD-566384.2
102.6
32.5
56.4
23.3
152.3
24.9
106.6
8.2
117.1
47.3
106.2
5.8
165.7
8.9


AD-1069890.1
90.0
63.7
39.5
10.3
87.7
45.3
106.1
4.2
93.1
52.3
95.1
8.7
105.1
54.2


AD-1069891.1
90.9
8.3
171.8
27.6
76.4
30.6
1.0
0.1
2.3
0.6
16.1
4.7
91.6
36.7


AD-1069892.1
87.7
4.3
103.5
64.9
53.3
18.4
10.9
1.2
16.3
4.9
76.7
10.6
63.9
22.0


AD-566388.2
89.3
9.3
81.8
44.2
66.4
8.8
83.9
2.8
104.1
12.5
106.4
14.7
79.5
10.5


AD-566389.1
96.8
9.7
133.1
83.6
100.5
41.9
100.0
2.6
118.4
17.9
107.1
9.6
120.4
50.2


AD-1069893.1
115.3
17.1
134.5
63.5
83.7
22.4
7.9
1.3
20.2
4.9
86.1
4.8
100.3
26.8


AD-566391.1
100.5
19.3
83.1
26.3
123.2
11.8
8.6
1.8
29.4
15.5
90.0
2.9
147.7
14.1


AD-1069894.1
111.0
9.5
59.6
20.7
192.8
78.0
106.5
4.6
122.2
24.5
118.5
11.2
156.3
12.2


AD-566393.1
71.5
41.2
71.8
14.8
63.8
27.2
86.1
13.1
87.3
38.2
106.8
14.7
76.4
32.5


AD-566395.1
129.1
44.8
144.5
35.2
63.7
19.1
21.6
4.9
48.8
6.6
96.4
11.9
76.4
22.8


AD-1069896.1
111.7
16.2
163.6
31.1
59.1
11.9
90.8
5.7
98.9
9.8
96.4
14.4
70.8
14.3


AD-1069897.1
117.2
27.4
165.7
34.2
56.6
27.0
66.1
7.2
83.6
11.4
106.0
5.2
67.8
32.4


AD-1069898.1
113.9
33.1
149.8
6.0
74.2
29.1
62.8
5.7
89.4
15.6
117.9
13.6
88.9
34.9


AD-1069899.1
110.5
23.1
140.1
17.7
130.9
46.9
121.6
12.5
139.5
29.2
114.8
14.1
156.8
56.2


AD-566475.1
113.8
39.3
84.2
68.5
38.1
4.0
28.3
6.6
18.5
5.7
48.2
8.6
45.6
4.8


AD-1069900.1
138.2
41.4
139.2
111.8
38.0
6.4
14.9
1.4
23.1
5.8
61.6
12.6
45.5
7.7


AD-566477.1
143.3
15.3
109.6
36.4
45.1
3.5
61.2
7.5
63.5
3.8
84.9
3.7
54.0
4.2


AD-1069901.1
119.6
12.0
146.5
66.0
81.6
32.7
102.3
7.2
94.4
6.6
102.2
8.9
97.8
39.2


AD-566483.1
93.8
37.5
128.6
19.1
71.1
3.8
10.7
0.8
14.7
1.1
85.8
1.9
85.2
4.6


AD-566484.1
113.5
27.1
146.1
26.1
68.7
24.4
32.3
13.4
51.8
10.0
99.5
6.7
82.3
29.3


AD-566485.2
127.7
22.8
173.0
16.5
85.3
37.7
106.1
13.3
115.6
11.4
119.0
14.5
102.2
45.2


AD-566486.1
60.4
20.1
107.2
56.5
142.2
73.2
50.8
9.3
58.7
21.8
117.6
5.2
170.3
87.7


AD-1069902.1
53.3
36.3
40.6
19.3
95.8
75.0
0.2
0.1
0.6
0.2
0.8
0.1
114.8
89.9


AD-1069903.1
88.0
13.3
148.8
55.5
104.6
55.8
35.7
7.6
40.5
14.3
76.5
9.7
125.3
66.8


AD-1069904.1
130.4
21.0
99.7
41.0
87.1
16.1
15.7
3.4
21.0
8.0
73.2
11.8
104.4
19.3


AD-1069905.1
109.9
4.2
117.9
52.4
85.6
18.6
91.2
3.4
88.3
13.6
97.5
10.5
102.6
22.3


AD-567054.1
152.7
15.6
163.6
99.0
89.5
20.5
71.2
12.9
74.5
12.7
92.6
8.3
107.3
24.5


AD-1069906.1
123.5
13.0
129.7
18.6
83.1
33.4
107.6
12.1
97.8
7.9
99.0
8.2
99.6
40.0


AD-1069907.1
126.7
33.5
132.0
74.4
89.0
55.4
114.9
4.0
109.7
10.3
99.4
8.5
106.6
66.4


AD-567057.1
95.7
30.3
81.3
36.2
132.1
53.1
36.1
15.0
59.5
12.2
96.4
10.8
201.8
101.3


AD-1069908.1
100.1
37.9
61.6
42.8
70.9
33.2
110.1
22.4
95.6
20.3
97.2
8.1
85.0
39.8


AD-567059.1
107.3
7.7
79.7
43.1
51.4
19.8
115.4
9.9
123.4
16.4
119.3
19.4
61.6
23.7


AD-567060.1
87.9
23.1
78.4
19.8
51.0
19.9
1.0
0.3
2.3
0.6
26.4
14.1
61.1
23.8


AD-1069909.1
134.1
20.5
84.9
47.6
83.2
32.7
10.0
1.8
11.0
2.3
61.0
11.8
99.7
39.2


AD-1069910.1
124.2
30.7
147.1
56.9
42.3
8.2
67.7
12.3
69.6
12.2
102.1
20.2
50.7
9.8


AD-567063.4
90.6
13.4
48.6
19.4
62.0
15.1
1.4
0.2
2.2
0.3
35.0
17.6
74.3
18.1


AD-1069911.1
47.8
13.8
86.7
84.5
85.3
19.5
1.2
0.1
2.7
1.0
21.4
5.4
102.2
23.3


AD-567065.1
37.1
5.3
90.8
70.5
54.6
30.8
131.2
8.1
125.6
29.8
124.4
21.7
65.4
36.9


AD-567066.4
103.1
16.9
34.6
21.0
53.5
13.2
0.3
0.0
0.7
0.3
7.1
2.2
64.1
15.8


AD-1069912.1
108.4
37.9
112.5
34.5
48.0
10.4
0.5
0.2
0.8
0.2
9.1
4.1
57.5
12.4


AD-567068.1
112.7
20.5
112.5
13.4
52.1
17.9
39.8
11.2
23.4
2.9
85.7
7.7
62.5
21.5


AD-1069913.1
120.3
31.4
110.3
24.4
40.2
6.5
9.9
2.2
36.0
8.4
111.0
26.6
48.2
7.8


AD-567070.1
120.6
26.5
125.7
68.0
46.4
6.9
1.6
0.5
5.8
2.1
93.4
31.1
55.6
8.3


AD-1069914.1
71.6
31.3
44.7
17.3
130.9
65.4
1.4
0.1
3.2
0.4
65.4
6.3
156.8
78.4


AD-567072.1
55.9
3.0
24.4
12.5
49.3
36.6
1.3
0.3
2.1
0.4
45.3
14.0
59.1
43.9


AD-1069915.1
93.1
25.8
44.0
6.5
60.3
24.8
0.8
0.1
1.2
0.2
10.5
1.6
72.3
29.8


AD-1069916.1
109.8
18.2
99.2
61.3
52.2
6.3
0.5
0.1
1.7
0.6
24.8
10.7
62.6
7.5


AD-1069917.1
118.8
32.2
99.0
27.5
45.1
9.0
0.3
0.2
0.5
0.1
2.5
0.8
54.0
10.8


AD-567076.1
99.5
4.2
115.5
92.3
38.2
9.8
90.9
6.3
59.2
4.7
105.0
8.1
45.7
11.8


AD-1069918.1
112.6
32.1
64.0
28.4
59.4
17.1
92.3
21.7
88.7
22.5
141.0
6.0
71.2
20.5


AD-567294.1
134.9
38.6
115.6
63.1
64.7
50.6
111.9
15.5
71.9
23.0
117.9
29.5
77.6
60.7


AD-1069919.1
111.9
46.9
54.8
32.3
118.7
25.9
2.0
0.2
2.3
0.4
34.4
18.1
142.3
31.0


AD-1069920.1
53.1
13.4
31.1
20.3
115.6
34.6
0.4
0.2
0.7
0.1
2.0
0.5
138.5
41.5


AD-567297.1
87.3
54.6
81.8
52.4
100.8
56.5
4.0
0.3
7.9
1.7
86.4
21.6
120.7
67.7


AD-567300.1
78.0
51.8
22.7
10.4
54.6
27.1
0.5
0.3
0.4
0.3
1.5
0.3
65.5
32.5


AD-567301.1
47.1
29.1
43.7
34.7
65.6
20.4
0.4
0.3
0.4
0.2
0.8
0.1
78.6
24.5


AD-1069922.1
57.0
44.0
36.7
17.3
131.5
51.5
0.7
0.3
0.7
0.2
10.3
5.8
157.5
61.7






#Transfection (TX)



*Free Uptake (FU)













TABLE 29







C3 Single Dose Screens in PCH cells (% C3 mRNA Remaining)






















FU*

FU*

FU*

TX#

TX#

TX#

TX#



Duplex ID
500 nM
STDEV
100 nM
STDEV
10 nM
STDEV
50 nM
STDEV
10 nM
STDEV
1 nM
STDEV
0.1 nM
STDEV
























AD-1069923.1
116.2
27.1
81.4
38.3
179.0
108.7
2.6
0.6
1.3
0.2
47.0
14.8
119.9
20.1


AD-1069924.1
102.9
34.6
33.8
13.8
168.2
33.6
1.8
0.6
0.4
0.1
39.7
33.6
64.8
21.9


AD-567305.1
130.3
31.6
72.2
8.2
291.8
126.6
7.8
1.5
1.7
0.6
36.2
23.0
127.1
21.7


AD-567306.1
134.9
46.0
137.7
42.7
257.1
17.6
84.2
46.0
39.0
21.4
136.0
15.9
226.3
31.8


AD-567308.1
93.4
38.5
146.2
55.5
278.5
63.2
1.0
0.6
0.8
0.1
48.7
20.7
112.7
23.6


AD-567309.1
162.5
19.0
295.9
146.1
276.8
114.1
2.0
0.7
1.2
0.2
56.0
9.4
126.8
26.0


AD-1069925.1
99.8
18.2
444.7
194.1
N/A
N/A
1.4
0.2
0.9
0.2
68.8
30.9
128.0
33.1


AD-567311.1
96.5
11.9
118.8
62.2
N/A
N/A
0.7
0.2
0.4
0.1
12.6
12.0
123.3
55.4


AD-567312.1
102.6
33.6
90.9
62.9
N/A
N/A
1.8
0.5
1.2
0.4
35.0
7.7
79.9
39.9


AD-1069926.1
116.0
29.4
97.5
22.9
209.7
100.9
70.8
37.0
25.9
0.7
96.2
30.6
155.8
60.7


AD-567314.2
150.3
39.8
138.4
74.7
201.6
86.9
45.4
33.7
36.8
4.5
126.3
52.9
182.0
9.8


AD-567315.6
52.5
11.9
48.1
6.4
162.4
136.5
4.0
3.5
0.3
0.1
81.0
36.4
20.0
5.2


AD-1069927.1
83.4
13.0
92.8
33.3
200.9
85.4
1.7
1.2
0.3
0.0
100.8
74.2
33.0
11.0


AD-1069928.1
209.2
48.7
167.8
79.2
176.4
74.2
30.5
6.1
13.1
3.3
N/A
N/A
159.7
26.3


AD-567318.2
201.4
24.6
206.8
55.7
173.8
78.7
1.2
0.3
1.3
0.3
49.2
4.6
162.1
40.8


AD-567319.1
218.4
46.8
173.7
12.9
281.0
91.9
5.3
0.5
5.4
1.5
144.3
55.0
190.7
24.4


AD-1069929.1
94.1
20.6
47.2
16.8
192.4
134.2
5.0
0.4
4.9
1.0
65.1
7.7
96.6
14.2


AD-567321.1
59.5
14.9
40.7
7.4
79.3
28.8
0.8
0.4
0.2
0.0
118.7
24.8
25.9
9.4


AD-1069930.1
95.1
15.5
100.1
17.5
161.3
37.9
1.7
0.5
0.4
0.1
50.8
27.2
68.7
18.7


AD-567323.1
204.7
17.6
165.4
35.3
203.3
62.1
24.8
4.1
14.0
0.8
126.1
26.8
273.1
54.0


AD-1069931.1
187.8
63.2
110.9
15.3
205.8
39.3
147.6
30.3
52.6
11.1
144.0
19.4
223.7
57.8


AD-567325.1
205.1
62.3
148.1
50.0
185.1
131.7
0.5
0.1
0.3
0.0
99.9
88.7
61.0
10.9


AD-567326.1
207.1
17.8
144.3
25.6
295.7
95.1
1.0
0.2
0.8
0.1
38.3
15.3
163.1
48.5


AD-1069932.1
80.4
19.5
128.3
145.9
147.4
119.4
34.0
9.0
19.3
3.2
63.0
21.7
56.2
6.0


AD-1069933.1
52.4
11.3
57.7
30.2
130.5
80.5
0.7
0.3
0.7
0.2
69.1
55.9
41.7
15.8


AD-567479.1
108.0
16.6
118.1
78.7
181.8
40.0
172.2
28.4
75.2
9.7
141.3
23.7
157.2
17.8


AD-567480.1
152.0
52.8
92.5
32.3
194.7
150.3
6.1
0.4
4.7
1.1
143.6
77.6
194.7
65.9


AD-567481.1
161.6
33.7
94.1
32.1
107.2
26.9
0.8
0.1
0.7
0.1
52.6
33.1
188.8
16.8


AD-567482.1
209.4
24.5
150.6
45.5
152.3
49.1
1.2
0.2
1.5
0.3
99.2
101.0
211.0
53.0


AD-1069934.1
272.6
45.2
154.1
47.2
246.4
98.2
188.8
17.8
22.8
1.9
149.2
64.6
226.5
25.9


AD-567485.1
55.6
16.2
84.6
5.0
190.1
49.6
8.5
1.7
8.7
2.4
68.3
4.3
51.4
12.9


AD-1069935.1
29.7
10.2
76.8
31.4
146.3
81.9
2.2
2.9
0.6
0.3
107.9
54.2
42.6
2.8


AD-567487.2
61.0
21.7
66.7
21.8
67.7
12.1
3.0
2.6
1.3
0.5
64.8
37.5
66.9
17.4


AD-567488.1
73.9
16.2
70.9
19.5
118.8
20.7
4.1
2.8
1.5
0.4
49.5
5.8
94.5
13.3


AD-567489.1
78.5
30.2
97.2
43.8
140.2
60.7
1.8
0.1
1.2
0.3
45.1
28.9
138.1
14.4


AD-1069936.1
104.9
39.1
75.3
24.7
158.3
53.1
0.4
0.1
0.3
0.0
155.1
111.3
48.3
9.0


AD-567491.1
154.0
28.5
101.3
61.1
166.5
98.7
0.7
0.1
0.8
0.2
56.9
9.5
143.5
42.2


AD-1069937.1
231.3
16.6
142.1
68.6
222.8
142.7
218.9
18.1
113.8
33.9
212.4
20.9
204.5
19.0


AD-1069938.1
46.5
13.8
55.3
4.6
150.7
116.5
129.1
46.8
51.9
10.0
96.3
50.3
69.1
17.4


AD-1069939.1
50.0
28.4
107.4
98.8
123.4
46.8
47.8
16.1
38.4
8.7
88.2
33.0
61.4
8.6


AD-567513.1
53.5
11.7
114.5
58.8
77.7
18.3
28.3
5.4
29.5
7.6
81.6
10.8
84.9
10.4


AD-567514.1
110.4
17.7
84.9
27.5
101.1
18.2
121.5
31.7
84.7
35.5
128.7
41.6
116.6
25.6


AD-1069940.1
70.4
12.5
73.1
21.7
169.2
31.9
22.2
3.1
13.9
1.6
83.3
7.0
132.4
20.2


AD-1069941.1
72.5
21.3
53.9
23.9
144.0
86.9
0.7
0.1
0.5
0.1
56.0
28.1
73.8
4.9


AD-1069942.1
69.5
27.5
43.0
11.4
119.8
15.8
0.5
0.1
0.6
0.1
92.6
61.0
77.9
28.3


AD-567518.1
38.6
23.2
80.0
4.3
112.2
111.5
89.7
27.7
31.9
6.7
83.5
25.1
58.4
10.1


AD-1069943.1
53.2
27.4
84.1
61.3
124.4
65.2
36.5
7.1
17.1
1.6
86.1
8.6
124.5
29.2


AD-567521.4
35.0
26.6
37.9
16.1
69.5
34.0
1.2
0.7
0.7
0.2
86.7
81.2
63.5
20.0


AD-1069944.1
44.8
19.9
52.6
33.8
81.9
24.9
7.0
0.6
6.6
0.5
60.9
23.9
120.0
18.3


AD-567524.1
89.2
59.0
53.1
27.4
99.6
99.2
5.4
1.3
6.5
2.2
113.1
80.0
165.8
19.3


AD-567525.1
62.6
11.3
53.3
18.8
109.7
43.2
1.2
0.2
1.9
0.3
59.6
11.6
124.2
27.2


AD-1069945.1
133.9
32.5
82.1
25.0
177.8
59.9
21.4
2.5
58.8
17.4
94.2
23.7
146.4
13.4


AD-567527.1
93.2
34.2
35.1
33.7
145.0
49.3
0.7
0.1
0.9
0.1
49.6
46.8
73.4
4.6


AD-1069946.1
44.2
10.6
40.8
9.0
134.5
123.5
0.6
0.1
0.5
0.1
51.1
18.8
27.7
6.4


AD-567529.1
48.1
6.9
N/A
N/A
74.9
25.7
1.9
1.2
1.2
0.1
48.1
33.4
67.6
28.4


AD-1069947.1
68.7
18.7
50.0
12.6
67.1
16.7
10.4
3.0
10.0
1.0
59.1
13.6
116.6
22.7


AD-567531.1
56.7
6.7
38.2
6.9
57.9
25.5
2.2
2.9
0.5
0.1
100.1
109.3
23.0
3.6


AD-567532.1
94.8
8.0
65.5
25.1
201.4
115.2
0.7
0.1
0.5
0.1
133.0
2.0
71.5
19.5


AD-567533.1
109.5
23.7
57.4
19.2
105.8
12.8
25.8
6.1
31.9
18.2
140.0
65.8
148.8
50.5


AD-1069948.1
87.7
33.7
63.2
22.2
122.7
50.7
1.1
0.4
1.9
0.4
105.1
73.0
111.1
29.9


AD-567535.1
107.4
24.7
55.2
4.1
154.2
65.1
0.8
0.2
0.8
0.1
63.5
67.6
109.8
21.8


AD-568149.1
76.7
29.9
57.9
20.3
58.0
40.3
96.3
23.9
89.6
58.4
86.2
29.6
62.2
5.9


AD-568150.1
55.6
4.1
65.0
30.0
88.8
44.6
1.0
0.2
1.2
0.2
N/A
N/A
56.9
12.1


AD-1069949.1
65.9
11.6
57.3
8.4
83.2
61.3
1.2
0.2
4.2
5.5
61.2
22.8
67.1
2.7


AD-1069950.1
89.1
22.9
52.0
10.2
102.4
23.6
8.5
0.8
10.8
3.1
101.9
29.2
120.3
32.2


AD-1069951.1
110.2
10.9
57.9
15.5
104.6
12.0
2.1
0.9
1.5
0.2
61.0
50.5
107.2
29.0


AD-1069952.1
104.0
36.7
60.5
31.8
132.8
45.4
1.4
0.4
2.1
1.1
99.1
34.4
72.6
14.3


AD-568155.1
78.2
29.3
92.1
48.5
143.8
40.2
102.3
46.2
163.3
39.9
97.2
30.8
124.1
18.2


AD-568159.1
52.2
13.2
33.4
10.4
119.0
39.7
99.5
39.5
95.0
26.4
99.9
26.8
86.1
12.5


AD-1069953.1
52.8
21.1
53.5
13.3
76.2
47.0
11.5
4.1
10.1
1.5
54.4
10.0
85.9
19.7


AD-568161.2
80.6
24.1
44.8
16.1
78.0
48.1
59.1
7.8
101.1
37.5
74.2
17.8
101.8
28.4


AD-568162.1
45.8
9.2
60.5
18.7
57.0
18.2
37.6
10.6
120.4
23.8
83.0
4.6
118.8
15.4


AD-1069954.1
64.2
20.8
32.8
8.3
71.4
20.9
106.8
36.7
116.0
7.8
98.4
27.9
108.5
35.8


AD-1069955.1
83.2
12.8
81.3
25.1
84.9
90.5
179.5
57.7
161.9
17.3
83.4
32.4
95.5
30.8


AD-568165.1
106.8
58.7
65.1
4.4
76.1
53.1
156.5
45.0
178.2
19.8
124.1
13.8
75.4
9.6


AD-1069956.1
29.1
6.4
31.0
15.9
58.9
48.9
2.2
1.1
0.5
0.1
8.7
12.1
24.7
6.0


AD-568337.1
60.8
22.7
38.8
16.8
59.7
26.6
123.3
24.8
120.7
24.2
100.5
21.7
71.4
18.6


AD-568338.1
86.4
7.9
38.1
14.8
60.2
35.7
6.9
1.4
7.5
3.6
78.3
76.1
77.8
18.7


AD-1069957.1
59.9
32.6
40.9
18.8
53.5
16.2
2.4
1.0
2.2
0.2
21.2
23.7
67.8
6.1


AD-568340.1
56.9
24.4
39.4
10.5
99.2
53.1
1.9
0.4
3.1
1.5
130.6
49.3
90.6
11.3


AD-1069958.1
40.3
14.3
55.5
10.7
55.6
5.5
0.9
0.3
0.8
0.1
N/A
N/A
40.4
2.2


AD-568342.1
121.1
27.0
65.7
14.4
144.7
49.6
71.1
33.5
140.9
19.0
87.9
23.1
101.0
38.5


AD-568343.4
81.1
15.2
43.1
9.6
75.7
40.2
2.6
0.4
3.3
0.8
67.7
69.3
71.4
9.8


AD-1069959.1
54.5
14.6
45.4
41.0
85.8
65.3
2.0
0.7
4.8
0.2
50.9
16.4
73.3
29.2


AD-568345.2
70.6
13.7
33.2
9.7
98.8
48.3
2.1
0.8
2.0
0.2
61.2
29.8
74.6
25.6


AD-568348.1
59.8
10.6
52.4
19.7
76.7
61.3
23.5
15.5
67.3
15.4
82.0
32.7
57.9
13.2


AD-1069961.1
38.0
11.4
71.2
21.8
98.5
32.0
0.8
0.3
0.9
0.1
122.3
50.1
44.5
12.5






#Transfection (TX)



*Free Uptake (FU)






Example 5. Structure-Activity Relationship Analyses

Based on the in vitro analyses in Example 4, structure-active relationship (SAR) analyses were performed. In particular, additional duplexes were designed, synthesized, and assayed in vitro.


siRNAs were synthesized and annealed using routine methods known in the art and described above.


Detailed lists of the unmodified commmplement component C3 sense and antisense strand nucleotide sequences are shown in Table 30. Detailed lists of the modified complement component C3 sense and antisense strand nucleotide sequences are shown in Table 31.


Free uptake experiments and transfection experiments in primary cynomolgu hepatocytes (PCH) were performed as described above.


Single dose free uptake experiments were performed at 500 nM, 100 nM, 10 nM, and 1 nM final duplex concentration.


Single dose transfection experiments were performed at 50 nM, 10 nM, 1 nM, and 0.1 nM final duplex concentration.


The results of the free uptake experiments are shown in Table 32 and the results of the transfection assays are shown in Table 33.









TABLE 30







Unmodified Sense and Antisense Strand Sequences of Complement Component C3 dsRNA Agents















SEQ


SEQ




Sense
ID
Range in
Antisense
ID
Range in


Duplex Name
Sequence 5’ to 3’
NO:
NM_000064.3
Sequence 5’ to 3’
NO:
NM_000064.3





AD-564742.5
CCAGACAGACAAGACCAUCUU
3758
 489-509
AAGAUGGUCUUGUCUGUCUGGAU
3937
 487-509





AD-1181478.1
CCAGACAGACAAGACCAUCUU
3759
 489-509
AAGATGGUCUUGUCUGUCUGGAU
3938
 487-509





AD-1181479.1
CCAGACAGACAAGACCAUCUU
3760
 489-509
AAGATGGUCUUGUCUGUCUGGAU
3939
 487-509





AD-1181480.1
CCAGACAGACAAGACCAUCUU
3761
 489-509
AAGATGGUCUUGUCUGUCUGGCU
3940
 487-509





AD-1181481.1
CCAGACAGACAAGACCAUCUU
3762
 489-509
AAGATGGUCUUGUCUGUCUGGCC
3941
 487-509





AD-1181482.1
AGACAGACAAGACCAUCUU
3763
 491-509
AAGATGGUCUUGUCUGUCUGG
3942
 489-509





AD-1181483.1
CCAGACAGACAAGACCAUCUU
3764
 489-509
AAGATGGUCUUGUCUGUCUGGCU
3943
 487-509





AD-1181484.1
CCAGACAGACAAGACCAUCUU
3765
 489-509
AAGATGGUCUUGUCUGUCUGGCU
3944
 487-509





AD-567304.4
GACUUCCUUGAAGCCAACUAU
3766
3613-3633
AUAGUUGGCUUCAAGGAAGUCUC
3945
3611-3633





AD-1181485.1
GACUUCCUUGAAGCCAACUAU
3767
3613-3633
AUAGTUGGCUUCAAGGAAGUCUC
3946
3611-3633





AD-1181486.1
GACUUCCUUGAAGCCAACUAU
3768
3613-3633
AUAGTUGGCUUCAAGGAAGUCCC
3947
3611-3633





AD-1181487.1
GACUUCCUUGAAGCCAACUAU
3769
3613-3633
AUAGTUGGCUUCAAGGAAGUCCC
3948
3611-3633





AD-1181488.1
CUUCCUUGAAGCCAACUAU
3770
3615-3633
AUAGTUGGCUUCAAGGAAGUC
3949
3613-3633





AD-1181489.1
GACUUCCUTGAAGCCAACUAU
3771
3613-3633
AUAGTUGGCUUCAAGGAAGUCCC
3950
3611-3633





AD-1181490.1
GACUUCCUTGAAGCCAACUAU
3772
3613-3633
AUAGTUGGCUUCAAGGAAGUCCC
3951
3611-3633





AD-1181491.1
GACUUCCUTGAAGCCAACUAU
3773
3613-3633
AUAGTUGGCUUCAAGGAAGUCCC
3952
3611-3633





AD-1181492.1
GACUUCCUTGAAGCCAACUAU
3774
3613-3633
AUAGTUGGCUUCAAGGAAGUCCC
3953
3611-3633





AD-567315.8
AGCCAACUACAUGAACCUACU
3775
3624-3644
AGUAGGTUCAUGUAGUUGGCUUC
3954
3622-3644





AD-1181493.1
AGCCAACUACAUGAACCUACU
3776
3624-3644
AGUAGGTUCAUGUAGUUGGCUUC
3955
3622-3644





AD-1181494.1
AGCCAACUACAUGAACCUACU
3777
3624-3644
AGUAGGTUCAUGUAGUUGGCUUC
3956
3622-3644





AD-1181495.1
AGCCAACUACAUGAACCUACU
3778
3624-3644
AGUAGGTUCAUGUAGUUGGCUUC
3957
3622-3644





AD-1181496.1
AGCCAACUACAUGAACCUACU
3779
3624-3644
AGUAGGTUCAUGUAGUUGGCUCC
3958
3622-3644





AD-1181497.1
CCAACUACAUGAACCUACU
3780
3626-3644
AGUAGGTUCAUGUAGUUGGCU
3959
3624-3644





AD-1181498.1
AGCCAACUACAUGAACCUACU
3781
3624-3644
AGUAGGTUCAUGUAGUUGGCUCC
3960
3622-3644





AD-1181499.1
AGCCAACUACAUGAACCUACU
3782
3624-3644
AGUAGGTUCAUGUAGUUGGCUCC
3961
3622-3644





AD-1181500.1
AGCCAACUACAUGAACCUACU
3783
3624-3644
AGUAGGUUCAUGUAGUUGGCUCC
3962
3622-3644





AD-1181501.1
AGCCAACUACAUGAACCUACU
3784
3624-3644
AGUAGGTUCAUGUAGUUGGCUCC
3963
3622-3644





AD-1181502.1
AGCCAACUACAUGAACCUACU
3785
3624-3644
AGUAGGTUCAUGUAGUUGGCUUC
3964
3622-3644





AD-568586.5
GAGAACCAGAAACAAUGCCAU
3786
5014-5034
AUGGCATUGUUUCUGGUUCUCUU
3965
5012-5034





AD-1181503.1
GAGAACCAGAAACAAUGCCAU
3787
5014-5034
AUGGCATUGUUUCUGGUUCUCUU
3966
5012-5034





AD-1181504.1
GAGAACCAGAAACAAUGCCAU
3788
5014-5034
AUGGCATUGUUUCUGGUUCUCUU
3967
5012-5034





AD-1181505.1
GAGAACCAGAAACAAUGCCAU
3789
5014-5034
AUGGCATUGUUUCUGGUUCUCCU
3968
5014-5034





AD-1181506.1
GAACCAGAAACAAUGCCAU
3790
5016-5034
AUGGCATUGUUUCUGGUUCUC
3969
5012-5034





AD-1181507.1
GAGAACCAGAAACAAUGCCAU
3791
5014-5034
ATGGCATUGUUUCUGGUUCUCCU
3970
5012-5034





AD-1181508.1
GAGAACCAGAAACAAUGCCAU
3792
5014-5034
AUGGCATUGUUUCUGGUUCUCCU
3971
5012-5034





AD-1181509.1
GAGAACCAGAAACAAUGCCAU
3793
5014-5034
AUGGCATUGUUUCUGGUUCUCCU
3972
5012-5034





AD-1181510.1
GAGAACCAGAAACAAUGCCAU
3794
5014-5034
AUGGCATUGUUUCUGGUUCUCCU
3973
5012-5034





AD-568978.5
ACAGACAAGACCAUCUACACU
3795
 493-513
AGUGUAGAUGGUCUUGUCUGUCU
3974
 491-513





AD-1181511.1
ACAGACAAGACCAUCUACACU
3796
 493-513
AGUGUAGAUGGUCUUGUCUGUCU
3975
 491-513





AD-1181513.1
ACAGACAAGACCAUCUACACU
3797
 493-513
AGUGTAGAUGGUCUUGUCUGUGC
3976
 491-513





AD-1181514.1
AGACAAGACCAUCUACACU
3798
 495-513
AGUGTAGAUGGUCUUGUCUGU
3977
 493-513





AD-1181515.1
ACAGACAAGACCAUCUACACU
3799
 493-513
AGUGTAGAUGGUCUUGUCUGUCU
3978
 491-513





AD-1181516.1
ACAGACAAGACCAUCUACACU
3800
 493-513
AGUGTAGAUGGUCUUGUCUGUCU
3979
 491-513





AD-1181517.1
ACAGACAAGACCAUCUACACU
3801
 493-513
AGUGTAGAUGGUCUUGUCUGUCU
3980
 491-513





AD-569164.9
AGAUCCGAGCCUACUAUGAAU
3802
 707-727
AUUCAUAGUAGGCUCGGAUCUUC
3981
 705-727





AD-1181518.1
AGAUCCGAGCCUACUAUGAAU
3803
 707-727
AUUCAUAGUAGGCUCGGAUCUUC
3982
 705-727





AD-1181519.1
AGAUCCGAGCCUACUAUGAAU
3804
 707-727
AUUCAUAGUAGGCUCGGAUCUCC
3983
 705-727





AD-1181520.1
AUCCGAGCCUACUAUGAAU
3805
 709-727
AUUCAUAGUAGGCUCGGAUCU
3984
 707-727





AD-1181521.1
AGAUCCGAGCCUACUAUGAAU
3806
 707-727
AUUCAUAGUAGGCUCGGAUCUUC
3985
 705-727





AD-1181522.1
AGAUCCGAGCCUACUAUGAAU
3807
 707-727
AUUCAUAGUAGGCUCGGAUCUUC
3986
 705-727





AD-1181523.1
AGAUCCGAGCCUACUAUGAAU
3808
 707-727
AUUCAUAGUAGGCUCGGAUCUUC
3987
 705-727





AD-1181524.1
AGAUCCGAGCCUACUAUGAAU
3809
 707-727
AUUCAUAGUAGGCUCGGAUCUUC
3988
 705-727





AD-570712.3
CCGAGCCGUUCUCUACAAUUU
3810
2634-2654
AAAUUGUAGAGAACGGCUCGGAU
3989
2632-2654





AD-1181525.1
CCGAGCCGUUCUCUACAAUUU
3811
2634-2654
AAAUUGUAGAGAACGGCUCGGAU
3990
2632-2654





AD-1181526.1
CCGAGCCGUUCUCUACAAUUU
3812
2634-2654
AAAUUGUAGAGAACGGCUCGGAU
3991
2632-2654





AD-1181527.1
CCGAGCCGUUCUCUACAAUUU
3813
2634-2654
AAAUTGTAGAGAACGGCUCGGAU
3992
2632-2654





AD-1181528.1
CCGAGCCGUUCUCUACAAUUU
3814
2634-2654
AAAUTGTAGAGAACGGCUCGGAU
3993
2632-2654





AD-1181529.1
CCGAGCCGUUCUCUACAAUUU
3815
2634-2654
AAAUTGTAGAGAACGGCUCGGGC
3994
2632-2654





AD-1181530.1
GAGCCGUUCUCUACAAUUU
3816
2636-2654
AAAUTGTAGAGAACGGCUCGG
3995
2634-2654





AD-1181531.1
CCGAGCCGTUCUCUACAAUUU
3817
2634-2654
AAAUTGTAGAGAACGGCUCGGGC
3996
2632-2654





AD-1181532.1
CCGAGCCGTUCUCUACAAUUU
3818
2634-2654
AAAUTGTAGAGAACGGCUCGGGC
3997
2632-2654





AD-1181533.1
CCGAGCCGTUCUCUACAAUUU
3819
2634-2654
AAAUTGTAGAGAACGGCUCGGGC
3998
2632-2654





AD-570713.3
CGAGCCGUUCUCUACAAUUAU
3820
2635-2655
AUAAUUGUAGAGAACGGCUCGGA
3999
2633-2655





AD-1181534.1
CGAGCCGUUCUCUACAAUUAU
3821
2635-2655
AUAAUUGUAGAGAACGGCUCGGA
4000
2633-2655





AD-1181535.1
CGAGCCGUUCUCUACAAUUAU
3822
2635-2655
AUAAUUGUAGAGAACGGCUCGGA
4001
2633-2655





AD-1181536.1
CGAGCCGUUCUCUACAAUUAU
3823
2635-2655
AUAAUUGUAGAGAACGGCUCGGC
4002
2633-2655





AD-1181537.1
CGAGCCGUUCUCUACAAUUAU
3824
2635-2655
AUAATUGUAGAGAACGGCUCGGC
4003
2633-2655





AD-1181538.1
AGCCGUUCUCUACAAUUAU
3825
2637-2655
AUAATUGUAGAGAACGGCUCG
4004
2635-2655





AD-1181539.1
CGAGCCGUUCUCUACAAUUAU
3826
2635-2655
AUAAUUGUAGAGAACGGCUCGGC
4005
2633-2655





AD-1181540.1
CGAGCCGUUCUCUACAAUUAU
3827
2635-2655
AUAAUUGUAGAGAACGGCUCGGC
4006
2633-2655





AD-1181541.1
CGAGCCGUTCTCUACAAUUAU
3828
2635-2655
AUAAUUGUAGAGAACGGCUCGGC
4007
2633-2655





AD-1181542.1
CGAGCCGUTCTCUACAAUUAU
3829
2635-2655
AUAAUUGUAGAGAACGGCUCGGC
4008
2633-2655





AD-570714.4
GAGCCGUUCUCUACAAUUACU
3830
2636-2656
AGUAAUUGUAGAGAACGGCUCGG
4009
2634-2656





AD-1181543.1
GAGCCGUUCUCUACAAUUACU
3831
2636-2656
AGUAAUUGUAGAGAACGGCUCGG
4010
2634-2656





AD-1181544.1
GAGCCGUUCUCUACAAUUACU
3832
2636-2656
AGUAAUUGUAGAGAACGGCUCGG
4011
2634-2656





AD-1181545.1
GAGCCGUUCUCUACAAUUACU
3833
2636-2656
AGUAAUUGUAGAGAACGGCUCCU
4012
2634-2656





AD-1181546.1
GCCGUUCUCUACAAUUACU
3834
2638-2656
AGUAAUUGUAGAGAACGGCUC
4013
2636-2656





AD-1181547.1
GAGCCGUUCUCUACAAUUACU
3835
2636-2656
AGUAAUUGUAGAGAACGGCUCGG
4014
2634-2656





AD-1181548.1
GAGCCGUUCUCUACAAUUACU
3836
2636-2656
AGUAAUUGUAGAGAACGGCUCGG
4015
2634-2656





AD-1181549.1
GAGCCGTUCUCUACAAUUACU
3837
2636-2656
AGUAAUUGUAGAGAACGGCUCGG
4016
2634-2656





AD-571826.5
CAAGCCUUGGCUCAAUACCAU
3838
3922-3942
AUGGUAUUGAGCCAAGGCUUGGA
4017
3920-3942





AD-1181550.1
CAAGCCUUGGCUCAAUACCAU
3839
3922-3942
AUGGUAUUGAGCCAAGGCUUGGA
4018
3920-3942





AD-1181551.1
CAAGCCUUGGCUCAAUACCAU
3840
3922-3942
AUGGUAUUGAGCCAAGGCUUGGC
4019
3920-3942





AD-1181552.1
CAAGCCUUGGCUCAAUACCAU
3841
3922-3942
AUGGTATUGAGCCAAGGCUUGGC
4020
3920-3942





AD-1181553.1
AGCCUUGGCUCAAUACCAU
3842
3924-3942
AUGGTATUGAGCCAAGGCUUG
4021
3922-3942





AD-1181554.1
CAAGCCUUGGCUCAAUACCAU
3843
3922-3942
AUGGTATUGAGCCAAGGCUUGGC
4022
3920-3942





AD-1181555.1
CAAGCCUUGGCUCAAUACCAU
3844
3922-3942
AUGGTATUGAGCCAAGGCUUGGC
4023
3920-3942





AD-572040.6
ACUCACCUGUAAUAAAUUCGU
3845
4158-4178
ACGAAUUUAUUACAGGUGAGUUG
4024
4156-4178





AD-1181556.1
ACUCACCUGUAAUAAAUUCGU
3846
4158-4178
ACGAAUUUAUUACAGGUGAGUUG
4025
4156-4178





AD-1181557.1
ACUCACCUGUAAUAAAUUCGU
3847
4158-4178
ACGAAUTUAUUACAGGUGAGUUG
4026
4156-4178





AD-1181558.1
ACUCACCUGUAAUAAAUUCGU
3848
4158-4178
ACGAAUUUAUUACAGGUGAGUCC
4027
4156-4178





AD-1181559.1
UCACCUGUAAUAAAUUCGU
3849
4160-4178
ACGAAUUUAUUACAGGUGAGU
4028
4158-4178





AD-1181560.1
ACUCACCUGUAAUAAAUUCGU
3850
4158-4178
ACGAAUUUAUUACAGGUGAGUUG
4029
4156-4178





AD-1181561.1
ACUCACCUGUAAUAAAUUCGU
3851
4158-4178
ACGAAUUUAUUACAGGUGAGUUG
4030
4156-4178





AD-1181562.1
ACUCACCUGUAAUAAAUUCGU
3852
4158-4178
ACGAAUUUAUUACAGGUGAGUUG
4031
4156-4178





AD-1181560.2
ACUCACCUGUAAUAAAUUCGU
3853
4158-4178
ACGAAUUUAUUACAGGUGAGUUG
4032
4156-4178





AD-572110.5
GAUGCCAAGAACACUAUGAUU
3854
4228-4248
AAUCAUAGUGUUCUUGGCAUCCU
4033
4226-4248





AD-1181563.1
GAUGCCAAGAACACUAUGAUU
3855
4228-4248
AAUCAUAGUGUUCUUGGCAUCCU
4034
4226-4248





AD-1181564.1
GAUGCCAAGAACACUAUGAUU
3856
4228-4248
AAUCAUAGUGUUCUUGGCAUCCU
4035
4226-4248





AD-1181565.1
GAUGCCAAGAACACUAUGAUU
3857
4228-4248
AAUCAUAGUGUUCUUGGCAUCCU
4036
4226-4248





AD-1181566.1
GAUGCCAAGAACACUAUGAUU
3858
4228-4248
AAUCAUAGUGUUCUUGGCAUCGG
4037
4226-4248





AD-1181567.1
UGCCAAGAACACUAUGAUU
3859
4230-4248
AAUCAUAGUGUUCUUGGCAUC
4038
4228-4248





AD-1181568.1
GAUGCCAAGAACACUAUGAUU
3860
4228-4248
AAUCAUAGUGUUCUUGGCAUCCU
4039
4226-4248





AD-1181569.1
GAUGCCAAGAACACUAUGAUU
3861
4228-4248
AAUCAUAGUGUUCUUGGCAUCCU
4040
4226-4248





AD-1181570.1
GAUGCCAAGAACACUAUGAUU
3862
4228-4248
AAUCAUAGUGUUCUUGGCAUCCU
4041
4226-4248





AD-1181571.1
GAUGCCAAGAACACUAUGAUU
3863
4228-4248
AAUCAUAGUGUUCUUGGCAUCCU
4042
4226-4248





AD-1181572.1
GAUGCCAAGAACACUAUGAUU
3864
4228-4248
AAUCAUAGUGUUCUUGGCAUCCU
4043
4226-4248





AD-572387.6
UCAAGGUCUACGCCUAUUACU
3865
4523-4543
AGUAAUAGGCGUAGACCUUGACU
4044
4521-4543





AD-1181573.1
UCAAGGUCUACGCCUAUUACU
3866
4523-4543
AGUAAUAGGCGUAGACCUUGACU
4045
4521-4543





AD-1181574.1
UCAAGGUCUACGCCUAUUACU
3867
4523-4543
AGUAAUAGGCGUAGACCUUGACU
4046
4521-4543





AD-1181575.1
UCAAGGUCUACGCCUAUUACU
3868
4523-4543
AGUAAUAGGCGUAGACCUUGACC
4047
4521-4543





AD-1181576.1
AAGGUCUACGCCUAUUACU
3869
4525-4543
AGUAAUAGGCGUAGACCUUGA
4048
4523-4543





AD-1181577.1
UCAAGGUCUACGCCUAUUACU
3870
4523-4543
AGUAAUAGGCGUAGACCUUGACU
4049
4521-4543





AD-1181578.1
UCAAGGUCUACGCCUAUUACU
3871
4523-4543
AGUAAUAGGCGUAGACCUUGACU
4050
4521-4543





AD-1181579.1
UCAAGGUCUACGCCUAUUACU
3872
4523-4543
AGUAAUAGGCGUAGACCUUGACU
4051
4521-4543





AD-1181580.1
UCAAGGUCTACGCCUAUUACU
3873
4523-4543
AGUAAUAGGCGUAGACCUUGACU
4052
4521-4543





AD-1181581.1
UCAAGGUCTACGCCUAUUACU
3874
4523-4543
AGUAAUAGGCGUAGACCUUGACU
4053
4521-4543





AD-569272.6
AAUUCUACUACAUCUAUAACU
3875
 815-835
AGUUAUAGAUGUAGUAGAAUUUC
4054
 813-835





AD-1181582.1
AAUUCUACUACAUCUAUAACU
3876
 815-835
AGUUAUAGAUGUAGUAGAAUUUC
4055
 813-835





AD-1181583.1
AAUUCUACUACAUCUAUAACU
3877
 815-835
AGUUAUAGAUGUAGUAGAAUUUC
4056
 813-835





AD-1181584.1
AAUUCUACUACAUCUAUAACU
3878
 815-835
AGUUAUAGAUGTAGUAGAAUUUC
4057
 813-835





AD-1181585.1
AAUUCUACUACAUCUAUAACU
3879
 815-835
AGUUAUAGAUGUAGUAGAAUUGG
4058
 813-835





AD-1181586.1
AAUUCUACUACAUCUAUAACU
3880
 815-835
AGUUAUAGAUGTAGUAGAAUUGG
4059
 813-835





AD-1181587.1
AAUUCUACUACAUCUAUAACU
3881
 815-835
AGUUAUAGAUGUAGUAGAAUU
4060
 815-833





AD-1181588.1
AAUUCUACUACAUCUAUAACU
3882
 815-835
AGUUAUAGAUGTAGUAGAAUU
4061
 815-833





AD-1181589.1
AAUUCUACUACAUCUAUAACU
3883
 815-835
AGUUAUAGAUGUAGUAGAAUUUC
4062
 815-835





AD-1181590.1
AAUUCUACUACAUCUAUAACU
3884
 815-835
AGUUAUAGAUGUAGUAGAAUUGG
4063
 815-835





AD-1181591.1
AAUUCUACUACAUCUAUAACU
3885
 815-835
AGUUAUAGAUGTAGUAGAAUUGG
4064
 815-835





AD-1181592.1
AAUUCUACUACAUCUAUAACU
3886
 815-835
AGUUAUAGAUGUAGUAGAAUU
4065
 815-833





AD-1181593.1
AAUUCUACUACAUCUAUAACU
3887
 815-835
AGUUAUAGAUGTAGUAGAAUU
4066
 815-833





AD-565034.2
CAGAGAAAUUCUACUACAUCU
3888
 809-829
AGAUGUAGUAGAAUUUCUCUGUA
4067
 807-829





AD-1181594.1
CAGAGAAAUUCUACUACAUCU
3889
 809-829
AGAUGUAGUAGAAUUUCUCUGUC
4068
 807-829





AD-1181595.1
CAGAGAAAUUCUACUACAUCU
3890
 809-829
AGAUGUAGUAGAAUUUCUCUGUC
4069
 807-829





AD-565035.2
AGAGAAAUUCUACUACAUCUU
3891
 810-830
AAGAUGTAGUAGAAUUUCUCUGU
4070
 808-830





AD-1181596.1
AGAGAAAUUCUACUACAUCUU
3892
 810-830
AAGATGTAGUAGAAUUUCUCUGU
4071
 808-830





AD-1181597.1
AGAGAAAUUCUACUACAUCUU
3893
 810-830
AAGATGUAGUAGAAUUUCUCUGU
4072
 808-830





AD-1181598.1
AGAGAAAUUCUACUAUAUCUU
3894
 810-830
AAGATATAGUAGAAUUUCUCUGU
4073
 808-830





AD-565037.2
AGAAAUUCUACUACAUCUAUU
3895
 812-832
AAUAGATGUAGUAGAAUUUCUCU
4074
 810-832





AD-1181599.1
AGAAAUUCUACUACAUCUAUU
3896
 812-832
AAUAGATGUAGTAGAAUUUCUCU
4075
 810-832





AD-1181600.1
AGAAAUUCUACUACAUCUAUU
3897
 812-832
AAUAGAUGUAGTAGAAUUUCUCU
4076
 810-832





AD-1181601.1
AGAAAUUCUACUACAUCUAUU
3898
 812-832
AAUAGATGUAGTAGAAUUUCUCU
4077
 810-832





AD-567072.2
CAAGGUCUUCUCUCUGGCUGU
3899
3342-3362
ACAGCCAGAGAGAAGACCUUGAC
4078
3340-3362





AD-1181602.1
CAAGGUCUUCUCUCUGGCUGU
3900
3342-3362
ACAGCCAGAGAGAAGACCUUGGC
4079
3340-3362





AD-1181603.1
CAAGGUCUUCUCUCUGGCUGU
3901
3342-3362
ACAGCCAGAGAGAAGACCUUGGC
4080
3340-3362





AD-1181604.1
CAAGGUCUUCUCUCUGGCUGU
3902
3342-3362
ACAGCCAGAGAGAAGACCUUGGC
4081
3340-3362





AD-567300.2
AGGAGACUUCCUUGAAGCCAU
3903
3609-3629
AUGGCUTCAAGGAAGUCUCCUGC
4082
3607-3629





AD-1181605.1
AGGAGACUUCCUUGAAGCCAU
3904
3609-3629
AUGGCUTCAAGGAAGUCUCCUGC
4083
3607-3629





AD-1181606.1
AGGAGACUUCCUUGAAGCCAU
3905
3609-3629
AUGGCUUCAAGGAAGUCUCCUGC
4084
3607-3629





AD-567301.2
GGAGACUUCCUUGAAGCCAAU
3906
3610-3630
AUUGGCTUCAAGGAAGUCUCCUG
4085
3608-3630





AD-1181607.1
GGAGACUUCCUUGAAGCCAAU
3907
3610-3630
AUUGGCTUCAAGGAAGUCUCCUG
4086
3608-3630





AD-1181608.1
GGAGACUUCCUUGAAGCCAAU
3908
3610-3630
AUUGGCUUCAAGGAAGUCUCCUG
4087
3608-3630





AD-569262.2
CCUACAGAGAAAUUCUACUAU
3909
 805-825
AUAGUAGAAUUUCUCUGUAGGCU
4088
 803-825





AD-1181609.1
CCUACAGAGAAAUUCUACUAU
3910
 805-825
AUAGTAGAAUUUCUCUGUAGGCU
4089
 803-825





AD-1181610.1
CCUACAGAGAAAUUCUACUAU
3911
 805-825
AUAGTAGAAUUUCUCUGUAGGCU
4090
 803-825





AD-569265.2
ACAGAGAAAUUCUACUACAUU
3912
 808-828
AAUGUAGUAGAAUUUCUCUGUAG
4091
 806-828





AD-1181611.1
ACAGAGAAAUUCUACUACAUU
3913
 808-828
AAUGTAGUAGAAUUUCUCUGUGG
4092
 806-828





AD-1181612.1
ACAGAGAAAUUCUACUACAUU
3914
 808-828
AAUGTAGUAGAAUUUCUCUGUGG
4093
 806-828





AD-569268.2
GAGAAAUUCUACUACAUCUAU
3915
 811-831
AUAGAUGUAGUAGAAUUUCUCUG
4094
 809-831





AD-1181613.1
GAGAAAUUCUACUACAUCUAU
3916
 811-831
AUAGAUGUAGUAGAAUUUCUCUG
4095
 809-831





AD-1181614.1
GAGAAAUUCUACUACAUCUAU
3917
 811-831
AUAGAUGUAGUAGAAUUUCUCUG
4096
 809-831





AD-569269.2
AGAAAUUCUACUACAUCUAUU
3918
 812-832
AAUAGAUGUAGUAGAAUUUCUCU
4097
 810-832





AD-1181615.1
AGAAAUUCUACUACAUCUAUU
3919
 812-832
AAUAGAUGUAGTAGAAUUUCUCU
4098
 810-832





AD-1181616.1
AGAAAUUCUACUACAUCUAUU
3920
 812-832
AAUAGATGUAGTAGAAUUUCUCU
4099
 810-832





AD-569270.2
GAAAUUCUACUACAUCUAUAU
3921
 813-833
AUAUAGAUGUAGUAGAAUUUCUC
4100
 811-833





AD-1181617.1
GAAAUUCUACUACAUCUAUAU
3922
 813-833
AUAUAGAUGUAGUAGAAUUUCUC
4101
 811-833





AD-1181618.1
GAAAUUCUACUACAUCUAUAU
3923
 813-833
AUAUAGAUGUAGUAGAAUUUCUC
4102
 811-833





AD-570676.2
ACCCUACUCUGUUGUUCGAAU
3924
2598-2618
AUUCGAACAACAGAGUAGGGUAG
4103
2596-2618





AD-1181619.1
ACCCUACUCUGUUGUUCGAAU
3925
2598-2618
AUUCGAACAACAGAGUAGGGUGG
4104
2596-2618





AD-1181620.1
ACCCUACUCUGUUGUUCGAAU
3926
2598-2618
AUUCGAACAACAGAGUAGGGUGG
4105
2596-2618





AD-571304.2
CAAGGUCUUCUCUCUGGCUGU
3927
3342-3362
ACAGCCAGAGAGAAGACCUUGAC
4106
3340-3362





AD-1181604.2
CAAGGUCUUCUCUCUGGCUGU
3928
3342-3362
ACAGCCAGAGAGAAGACCUUGGC
4107
3340-3362





AD-1181621.1
CAAGGUCUUCUCUCUGGCUGU
3929
3342-3362
ACAGCCAGAGAGAAGACCUUGGC
4108
3340-3362





AD-1069946.2
UGGCUCAAUGAACAGAGAUAU
3930
3856-3876
AUAUCUCUGUUCAUUGAGCCAAC
4109
3854-3876





AD-1181622.1
UGGCUCAAUGAACAGAGAUAU
3931
3856-3876
AUAUCUCUGUUCAUUGAGCCAGC
4110
3854-3876





AD-1181623.1
UGGCUCAAUGAACAGAGAUAU
3932
3856-3876
AUAUCUCUGUUCAUUGAGCCAGC
4111
3854-3876





AD-1181624.1
UGGCUCAAUGAACAGAGAUAU
3933
3856-3876
AUAUCUCUGUUCAUUGAGCCAGC
4112
3854-3876





AD-1069956.2
GCUGAGGAGAAUUGCUUCAUU
3934
4633-4653
AAUGAAGCAAUUCUCCUCAGCAC
4113
4631-4653





AD-1181625.1
GCUGAGGAGAAUUGCUUCAUU
3935
4633-4653
AAUGAAGCAAUUCUCCUCAGCGC
4114
4631-4653





AD-1181626.1
GCUGAGGAGAAUUGCUUCAUU
3936
4633-4653
AAUGAAGCAAUUCUCCUCAGCGC
4115
4631-4653
















TABLE 31







Modified Sense and Antisense Strand Sequences of Complement Component C3 dsRNA Agents
















SEQ

SEQ

SEQ
SEQ




ID

ID
mRNA
ID
ID


Duplex Name
Sense Sequence 5’ to 3’
NO:
Antisense Sequence 5’ to 3’
NO:
Target Sequence 5’ to 3’
NO:
NO:





AD-564742.5
cscsagacAfgAfCfAfagaccaucuuL96
4116
asAfsgaug(Ggn)ucuuguCfuGfucuggsasu
4295
AUCCAGACAGACAAGACCAUCUA
4474






AD-1181478.1
cscsagacAfgAfCfAfagaccaucuuL96
4117
asAfsgadTg(G2p)ucuuguCfuGfucuggsasu
4296
AUCCAGACAGACAAGACCAUCUA
4475






AD-1181479.1
cscsagacAfgAfCfAfagaccaucuuL96
4118
asAfsgadTg(G2p)ucuuguCfudGucuggsasu
4297
AUCCAGACAGACAAGACCAUCUA
4476






AD-1181480.1
cscsagacAfgAfCfAfagaccaucuuL96
4119
asAfsgadTg(G2p)ucuuguCfudGucuggscsu
4298
AUCCAGACAGACAAGACCAUCUA
4477






AD-1181481.1
cscsagacAfgAfCfAfagaccaucuuL96
4120
asAfsgadTg(G2p)ucuuguCfudGucuggscsc
4299
AUCCAGACAGACAAGACCAUCUA
4478






AD-1181482.1
asgsacAfgAfCfAfagaccaucuuL96
4121
asAfsgadTg(G2p)ucuuguCfudGucusgsg
4300
AUCCAGACAGACAAGACCAUCUA
4479






AD-1181483.1
cscsagacagdAcdAagaccaucuuL96
4122
asdAsgadTg(G2p)ucuuguCfudGucuggscsu
4301
AUCCAGACAGACAAGACCAUCUA
4480






AD-1181484.1
cscsagacdAgdACfdAagaccaucuuL96
4123
asdAsgadTg(G2p)ucuuguCfudGucuggscsu
4302
AUCCAGACAGACAAGACCAUCUA
4481






AD-567304.4
gsascuucCfuUfGfAfagccaacuauL96
4124
asUfsaguu(Ggn)gcuucaAfgGfaagucsusc
4303
AUCCAGACAGACAAGACCAUCUA
4482






AD-1181485.1
gsascuucCfuUfGfAfagccaacuauL96
4125
asUfsagdTu(G2p)gcuucaAfgdGaagucsusc
4304
GAGACUUCCUUGAAGCCAACUAC
4483






AD-1181486.1
gsascuucCfuUfGfAfagccaacuauL96
4126
asUfsagdTu(G2p)gcuucaAfgdGaagucscsc
4305
GAGACUUCCUUGAAGCCAACUAC
4484






AD-1181487.1
gsascuucCfuUfGfAfagccaacuauL96
4127
asUfsagdTu(G2p)gcuucadAgdGaagucscsc
4306
GAGACUUCCUUGAAGCCAACUAC
4485






AD-1181488.1
csusucCfuUfGfAfagccaacuauL96
4128
asUfsagdTu(G2p)gcuucadAgdGaagsusc
4307
GAGACUUCCUUGAAGCCAACUAC
4486






AD-1181489.1
gsascuucCfudTgdAagccaacuauL96
4129
asUfsagdTu(G2p)gcuucaAfgdGaagucscsc
4308
GAGACUUCCUUGAAGCCAACUAC
4487






AD-1181490.1
gsascuucdCudTgdAagccaacuauL96
4130
asUfsagdTu(G2p)gcuucadAgdGaagucscsc
4309
GAGACUUCCUUGAAGCCAACUAC
4488






AD-1181491.1
gsascuucdCudTgdAAgccaacuauL96
4131
asUfsagdTu(G2p)gcuucadAgdGaagucscsc
4310
GAGACUUCCUUGAAGCCAACUAC
4489






AD-1181492.1
gsascuucdCudTgdAdAgccaacuauL96
4132
asUfsagdTu(G2p)gcuucadAgdGaagucscsc
4311
GAGACUUCCUUGAAGCCAACUAC
4490






AD-567315.8
asgsccaaCfuAfCfAfugaaccuacuL96
4133
asGfsuagg(Tgn)ucauguAfgUfuggcususc
4312
GAAGCCAACUACAUGAACCUACA
4491






AD-1181493.1
asgsccaaCfuAfCfAfugaaccuacuL96
4134
asdGsuagg(Tgn)ucauguAfgUfuggcususc
4313
GAAGCCAACUACAUGAACCUACA
4492






AD-1181494.1
asgsccaaCfuAfCfAfugaaccuacuL96
4135
asdGsuadGg(Tgn)ucauguAfgUfuggcususc
4314
GAAGCCAACUACAUGAACCUACA
4493






AD-1181495.1
asgsccaaCfuAfCfAfugaaccuacuL96
4136
asdGsuadGg(Tgn)ucaugudAgUfuggcususc
4315
GAAGCCAACUACAUGAACCUACA
4494






AD-1181496.1
asgsccaaCfuAfCfAfugaaccuacuL96
4137
asdGsuadGg(Tgn)ucaugudAgUfuggcuscsc
4316
GAAGCCAACUACAUGAACCUACA
4495






AD-1181497.1
cscsaaCfuAfCfAfugaaccuacuL96
4138
asdGsuadGg(Tgn)ucaugudAgUfuggscsu
4317
GAAGCCAACUACAUGAACCUACA
4496






AD-1181498.1
asgsccaaCfudAcdAugaaccuacuL96
4139
asdGsuadGg(Tgn)ucaugudAgUfuggcuscsc
4318
GAAGCCAACUACAUGAACCUACA
4497






AD-1181499.1
asgsccaadCudAcdAugaaccuacuL96
4140
asdGsuadGg(Tgn)ucaugudAgUfuggcuscsc
4319
GAAGCCAACUACAUGAACCUACA
4498






AD-1181500.1
asgsccaadCudAcdAugaaccuacuL96
4141
asdGsuadGg(U2p)ucaugudAgUfuggcuscsc
4320
GAAGCCAACUACAUGAACCUACA
4499






AD-1181501.1
asgsccaadCudAcdAUfgaaccuacuL96
4142
asdGsuadGg(Tgn)ucaugudAgUfuggcuscsc
4321
GAAGCCAACUACAUGAACCUACA
4500






AD-1181502.1
asgsccaadCudAcdAUfgaaccuacuL96
4143
asdGsuagg(Tgn)ucauguAfgUfuggcususc
4322
GAAGCCAACUACAUGAACCUACA
4501






AD-568586.5
gsasgaacCfaGfAfAfacaaugccauL96
4144
asUfsggca(Tgn)uguuucUfgGfuucucsusu
4323
AAGAGAACCAGAAACAAUGCCAG
4502






AD-1181503.1
gsasgaacCfaGfAfAfacaaugccauL96
4145
asUfsggdCa(Tgn)uguuucUfgdGuucucsusu
4324
AAGAGAACCAGAAACAAUGCCAG
4503






AD-1181504.1
gsasgaacCfaGfAfAfacaaugccauL96
4146
asUfsggCfa(Tgn)uguuucUfgdGuucucsusu
4325
AAGAGAACCAGAAACAAUGCCAG
4504






AD-1181505.1
gsasgaacCfaGfAfAfacaaugccauL96
4147
asUfsggdCa(Tgn)uguuucUfgdGuucucscsu
4326
AAGAGAACCAGAAACAAUGCCAG
4505






AD-1181506.1
gsasacCfaGfAfAfacaaugccauL96
4148
asUfsggdCa(Tgn)uguuucUfgdGuucsusc
4327
AAGAGAACCAGAAACAAUGCCAG
4506






AD-1181507.1
gsasgaacCfadGadAacaaugccauL96
4149
asdTsggdCa(Tgn)uguuucUfgdGuucucscsu
4328
AAGAGAACCAGAAACAAUGCCAG
4507






AD-1181508.1
gsasgaacdCadGadAacaaugccauL96
4150
asUfsggdCa(Tgn)uguuucUfgdGuucucscsu
4329
AAGAGAACCAGAAACAAUGCCAG
4508






AD-1181509.1
gsasgaacdCadGadAAcaaugccauL96
4151
asUfsggdCa(Tgn)uguuucUfgdGuucucscsu
4330
AAGAGAACCAGAAACAAUGCCAG
4509






AD-1181510.1
gsasgaacdCadGadAdAcaaugccauL96
4152
asUfsggdCa(Tgn)uguuucUfgdGuucucscsu
4331
AAGAGAACCAGAAACAAUGCCAG
4510






AD-568978.5
ascsagacAfaGfAfCfcaucuacacuL96
4153
asGfsuguAfgAfUfggucUfuGfucuguscsu
4332
AGACAGACAAGACCAUCUACACC
4511






AD-1181511.1
ascsagacAfaGfAfCfcaucuacacuL96
4154
asdGsuguAfgAfUfggucUfudGucuguscsu
4333
AGACAGACAAGACCAUCUACACC
4512






AD-1181513.1
ascsagacAfaGfAfCfcaucuacacuL96
4155
asdGsugdTadGauggucUfudGucugusgsc
4334
AGACAGACAAGACCAUCUACACC
4513






AD-1181514.1
asgsacAfaGfAfCfcaucuacacuL96
4156
asdGsugdTadGauggucUfudGucusgsu
4335
AGACAGACAAGACCAUCUACACC
4514






AD-1181515.1
ascsagacdAadGadCcaucuacacuL96
4157
asdGsugdTadGauggucUfudGucuguscsu
4336
AGACAGACAAGACCAUCUACACC
4515






AD-1181516.1
ascsagacdAadGadCCfaucuacacuL96
4158
asdGsugdTadGauggucUfudGucuguscsu
4337
AGACAGACAAGACCAUCUACACC
4516






AD-1181517.1
ascsagacdAadGaCfcaucuacacuL96
4159
asdGsugdTadGauggucUfudGucuguscsu
4338
AGACAGACAAGACCAUCUACACC
4517






AD-569164.9
asgsauccGfaGfCfCfuacuaugaauL96
4160
asUfsucaUfaGfUfaggcUfcGfgaucususc
4339
GAAGAUCCGAGCCUACUAUGAAA
4518






AD-1181518.1
asgsauccGfaGfCfCfuacuaugaauL96
4161
asUfsucaUfaguaggcUfcdGgaucususc
4340
GAAGAUCCGAGCCUACUAUGAAA
4519






AD-1181519.1
asgsauccGfaGfCfCfuacuaugaauL96
4162
asUfsucaUfaguaggcUfcdGgaucuscsc
4341
GAAGAUCCGAGCCUACUAUGAAA
4520






AD-1181520.1
asusccGfaGfCfCfuacuaugaauL96
4163
asUfsucaUfaguaggcUfcdGgauscsu
4342
GAAGAUCCGAGCCUACUAUGAAA
4521






AD-1181521.1
asgsauccdGagCfCfuacuaugaauL96
4164
asUfsucaUfaguaggcUfcdGgaucususc
4343
GAAGAUCCGAGCCUACUAUGAAA
4522






AD-1181522.1
asgsauccdGadGcdCuacuaugaauL96
4165
asUfsucaUfaguaggcUfcdGgaucususc
4344
GAAGAUCCGAGCCUACUAUGAAA
4523






AD-1181523.1
asgsauccdGagCfCfUfacuaugaauL96
4166
asUfsucaUfaguaggcUfcdGgaucususc
4345
GAAGAUCCGAGCCUACUAUGAAA
4524






AD-1181524.1
asgsauccdGagCfCfuacuaugaauL96
4167
asUfsucdAudAguaggcUfcdGgaucususc
4346
GAAGAUCCGAGCCUACUAUGAAA
4525






AD-570712.3
cscsgagcCfgUfUfCfucuacaauuuL96
4168
asAfsauuGfuAfGfagaaCfgGfcucggsasu
4347
AUCCGAGCCGUUCUCUACAAUUA
4526






AD-1181525.1
cscsgagcCfgUfUfCfucuacaauuuL96
4169
asAfsauuGfuagagaaCfgdGcucggsasu
4348
AUCCGAGCCGUUCUCUACAAUUA
4527






AD-1181526.1
cscsgagcCfgUfUfCfucuacaauuuL96
4170
asdAsauuGfuagagaaCfgdGcucggsasu
4349
AUCCGAGCCGUUCUCUACAAUUA
4528






AD-1181527.1
cscsgagcCfgUfUfCfucuacaauuuL96
4171
asdAsaudTgdTagagaaCfgdGcucggsasu
4350
AUCCGAGCCGUUCUCUACAAUUA
4529






AD-1181528.1
cscsgagcCfgUfUfCfucuacaauuuL96
4172
asAfsaudTgdTagagaaCfgdGcucggsasu
4351
AUCCGAGCCGUUCUCUACAAUUA
4530






AD-1181529.1
cscsgagcCfgUfUfCfucuacaauuuL96
4173
asAfsaudTgdTagagaaCfgdGcucggsgsc
4352
AUCCGAGCCGUUCUCUACAAUUA
4531






AD-1181530.1
gsasgcCfgUfUfCfucuacaauuuL96
4174
asAfsaudTgdTagagaaCfgdGcucsgsg
4353
AUCCGAGCCGUUCUCUACAAUUA
4532






AD-1181531.1
cscsgagcCfgdTudCucuacaauuuL96
4175
asdAsaudTgdTagagaaCfgdGcucggsgsc
4354
AUCCGAGCCGUUCUCUACAAUUA
4533






AD-1181532.1
cscsgagcCfgdTudCUfcuacaauuuL96
4176
asdAsaudTgdTagagaaCfgdGcucggsgsc
4355
AUCCGAGCCGUUCUCUACAAUUA
4534






AD-1181533.1
cscsgagcdCgdTudCUfcuacaauuuL96
4177
asdAsaudTgdTagagaaCfgdGcucggsgsc
4356
AUCCGAGCCGUUCUCUACAAUUA
4535






AD-570713.3
csgsagccGfuUfCfUfcuacaauuauL96
4178
asUfsaauUfgUfAfgagaAfcGfgcucgsgsa
4357
UCCGAGCCGUUCUCUACAAUUAC
4536






AD-1181534.1
csgsagccGfuUfCfUfcuacaauuauL96
4179
asUfsaauUfguagagaAfcdGgcucgsgsa
4358
UCCGAGCCGUUCUCUACAAUUAC
4537






AD-1181535.1
csgsagccGfuUfCfUfcuacaauuauL96
4180
asUfsaauUfguagagadAcdGgcucgsgsa
4359
UCCGAGCCGUUCUCUACAAUUAC
4538






AD-1181536.1
csgsagccGfuUfCfUfcuacaauuauL96
4181
asUfsaauUfguagagadAcdGgcucgsgsc
4360
UCCGAGCCGUUCUCUACAAUUAC
4539






AD-1181537.1
csgsagccGfuUfCfUfcuacaauuauL96
4182
asUfsaadTudGuagagadAcdGgcucgsgsc
4361
UCCGAGCCGUUCUCUACAAUUAC
4540






AD-1181538.1
asgsccGfuUfCfUfcuacaauuauL96
4183
asUfsaadTudGuagagadAcdGgcuscsg
4362
UCCGAGCCGUUCUCUACAAUUAC
4541






AD-1181539.1
csgsagccdGuUfCfUfcuacaauuauL96
4184
asUfsaauUfguagagaAfcdGgcucgsgsc
4363
UCCGAGCCGUUCUCUACAAUUAC
4542






AD-1181540.1
csgsagccdGuUfCfUfcuacaauuauL96
4185
asUfsaauUfguagagadAcdGgcucgsgsc
4364
UCCGAGCCGUUCUCUACAAUUAC
4543






AD-1181541.1
csgsagccdGudTcdTcuacaauuauL96
4186
asUfsaauUfguagagadAcdGgcucgsgsc
4365
UCCGAGCCGUUCUCUACAAUUAC
4544






AD-1181542.1
csgsagccdGudTcdTCfuacaauuauL96
4187
asUfsaauUfguagagadAcdGgcucgsgsc
4366
UCCGAGCCGUUCUCUACAAUUAC
4545






AD-570714.4
gsasgccgUfuCfUfCfuacaauuacuL96
4188
asGfsuaaUfuGfUfagagAfaCfggcucsgsg
4367
CCGAGCCGUUCUCUACAAUUACC
4546






AD-1181543.1
gsasgccgUfuCfUfCfuacaauuacuL96
4189
asdGsuaaUfuguagagAfaCfggcucsgsg
4368
CCGAGCCGUUCUCUACAAUUACC
4547






AD-1181544.1
gsasgccgUfuCfUfCfuacaauuacuL96
4190
asdGsuaaUfuguagagdAaCfggcucsgsg
4369
CCGAGCCGUUCUCUACAAUUACC
4548






AD-1181545.1
gsasgccgUfuCfUfCfuacaauuacuL96
4191
asdGsuaaUfuguagagdAaCfggcucscsu
4370
CCGAGCCGUUCUCUACAAUUACC
4549






AD-1181546.1
gscscgUfuCfUfCfuacaauuacuL96
4192
asdGsuaaUfuguagagdAaCfggcsusc
4371
CCGAGCCGUUCUCUACAAUUACC
4550






AD-1181547.1
gsasgccgUfudCudCuacaauuacuL96
4193
asdGsuaaUfuguagagdAaCfggcucsgsg
4372
CCGAGCCGUUCUCUACAAUUACC
4551






AD-1181548.1
gsasgccgUfudCudCUfacaauuacuL96
4194
asdGsuaaUfuguagagdAaCfggcucsgsg
4373
CCGAGCCGUUCUCUACAAUUACC
4552






AD-1181549.1
gsasgccgdTudCudCUfacaauuacuL96
4195
asdGsuaaUfuguagagdAaCfggcucsgsg
4374
CCGAGCCGUUCUCUACAAUUACC
4553






AD-571826.5
csasagccUfuGfGfCfucaauaccauL96
4196
asUfsgguAfuUfGfagccAfaGfgcuugsgsa
4375
UCCAAGCCUUGGCUCAAUACCAA
4554






AD-1181550.1
csasagccUfuGfGfCfucaauaccauL96
4197
asUfsgguAfuugagccdAadGgcuugsgsa
4376
UCCAAGCCUUGGCUCAAUACCAA
4555






AD-1181551.1
csasagccUfuGfGfCfucaauaccauL96
4198
asUfsgguAfuugagccdAadGgcuugsgsc
4377
UCCAAGCCUUGGCUCAAUACCAA
4556






AD-1181552.1
csasagccUfuGfGfCfucaauaccauL96
4199
asUfsggdTadTugagccdAadGgcuugsgsc
4378
UCCAAGCCUUGGCUCAAUACCAA
4557






AD-1181553.1
asgsccUfuGfGfCfucaauaccauL96
4200
asUfsggdTadTugagccdAadGgcususg
4379
UCCAAGCCUUGGCUCAAUACCAA
4558






AD-1181554.1
csasagccUfudGgdCucaauaccauL96
4201
asUfsggdTadTugagccdAadGgcuugsgsc
4380
UCCAAGCCUUGGCUCAAUACCAA
4559






AD-1181555.1
csasagccUfudGgdCUfcaauaccauL96
4202
asUfsggdTadTugagccdAadGgcuugsgsc
4381
UCCAAGCCUUGGCUCAAUACCAA
4560






AD-572040.6
ascsucacCfuGfUfAfauaaauucguL96
4203
asCfsgaaUfuUfAfuuacAfgGfugagususg
4382
CAACUCACCUGUAAUAAAUUCGA
4561






AD-1181556.1
ascsucacCfuGfUfAfauaaauucguL96
4204
asCfsgaaUfuuauuacdAgdGugagususg
4383
CAACUCACCUGUAAUAAAUUCGA
4562






AD-1181557.1
ascsucacCfuGfUfAfauaaauucguL96
4205
asCfsgadAudTuauuacdAgdGugagususg
4384
CAACUCACCUGUAAUAAAUUCGA
4563






AD-1181558.1
ascsucacCfuGfUfAfauaaauucguL96
4206
asCfsgaaUfuuauuacdAgdGugaguscsc
4385
CAACUCACCUGUAAUAAAUUCGA
4564






AD-1181559.1
uscsacCfuGfUfAfauaaauucguL96
4207
asCfsgaaUfuuauuacdAgdGugasgsu
4386
CAACUCACCUGUAAUAAAUUCGA
4565






AD-1181560.1
ascsucacCfudGudAauaaauucguL96
4208
asCfsgaaUfuuauuacdAgdGugagususg
4387
CAACUCACCUGUAAUAAAUUCGA
4566






AD-1181561.1
ascsucacCfudGudAdAuaaauucguL96
4209
asCfsgaaUfuuauuacdAgdGugagususg
4388
CAACUCACCUGUAAUAAAUUCGA
4567






AD-1181562.1
ascsucacCfudGudAAuaaauucguL96
4210
asCfsgaaUfuuauuacdAgdGugagususg
4389
CAACUCACCUGUAAUAAAUUCGA
4568






AD-1181560.2
ascsucacCfudGudAauaaauucguL96
4211
asCfsgaaUfuuauuacdAgdGugagususg
4390
CAACUCACCUGUAAUAAAUUCGA
4569






AD-572110.5
gsasugccAfaGfAfAfcacuaugauuL96
4212
asAfsucaUfaGfUfguucUfuGfgcaucscsu
4391
AGGAUGCCAAGAACACUAUGAUC
4570






AD-1181563.1
gsasugccAfaGfAfAfcacuaugauuL96
4213
asAfsucaUfaguguucUfudGgcaucscsu
4392
AGGAUGCCAAGAACACUAUGAUC
4571






AD-1181564.1
gsasugccAfaGfAfAfcacuaugauuL96
4214
asdAsucdAudAguguucUfudGgcaucscsu
4393
AGGAUGCCAAGAACACUAUGAUC
4572






AD-1181565.1
gsasugccAfaGfAfAfcacuaugauuL96
4215
asdAsucaUfaguguucUfudGgcaucscsu
4394
AGGAUGCCAAGAACACUAUGAUC
4573






AD-1181566.1
gsasugccAfaGfAfAfcacuaugauuL96
4216
asdAsucaUfaguguucUfudGgcaucsgsg
4395
AGGAUGCCAAGAACACUAUGAUC
4574






AD-1181567.1
usgsccAfaGfAfAfcacuaugauuL96
4217
asdAsucaUfaguguucUfudGgcasusc
4396
AGGAUGCCAAGAACACUAUGAUC
4575






AD-1181568.1
gsasugccAfadGadACfacuaugauuL96
4218
asAfsucaUfaguguucUfudGgcaucscsu
4397
AGGAUGCCAAGAACACUAUGAUC
4576






AD-1181569.1
gsasugccAfadGadAcacuaugauuL96
4219
asAfsucaUfaguguucUfudGgcaucscsu
4398
AGGAUGCCAAGAACACUAUGAUC
4577






AD-1181570.1
gsasugccdAadGadAcacuaugauuL96
4220
asAfsucaUfaguguucUfudGgcaucscsu
4399
AGGAUGCCAAGAACACUAUGAUC
4578






AD-1181571.1
gsasugccdAadGadACfacuaugauuL96
4221
asAfsucaUfaguguucUfudGgcaucscsu
4400
AGGAUGCCAAGAACACUAUGAUC
4579






AD-1181572.1
gsasugccdAadGadACfacuaugauuL96
4222
asdAsucaUfaguguucUfudGgcaucscsu
4401
AGGAUGCCAAGAACACUAUGAUC
4580






AD-572387.6
uscsaaggUfcUfAfCfgccuauuacuL96
4223
asGfsuaaUfaGfGfcguaGfaCfcuugascsu
4402
AGUCAAGGUCUACGCCUAUUACA
4581






AD-1181573.1
uscsaaggUfcUfAfCfgccuauuacuL96
4224
asdGsuaaUfaggcguaGfaCfcuugascsu
4403
AGUCAAGGUCUACGCCUAUUACA
4582






AD-1181574.1
uscsaaggUfcUfAfCfgccuauuacuL96
4225
asdGsuaaUfaggcguadGaCfcuugascsu
4404
AGUCAAGGUCUACGCCUAUUACA
4583






AD-1181575.1
uscsaaggUfcUfAfCfgccuauuacuL96
4226
asdGsuaaUfaggcguadGaCfcuugascsc
4405
AGUCAAGGUCUACGCCUAUUACA
4584






AD-1181576.1
asasggUfcUfAfCfgccuauuacuL96
4227
asdGsuaaUfaggcguadGaCfcuusgsa
4406
AGUCAAGGUCUACGCCUAUUACA
4585






AD-1181577.1
uscsaaggUfcUfaCfgccuauuacuL96
4228
asdGsuaaUfaggcguadGaCfcuugascsu
4407
AGUCAAGGUCUACGCCUAUUACA
4586






AD-1181578.1
uscsaaggUfcUfdACfgccuauuacuL96
4229
asdGsuaaUfaggcguadGaCfcuugascsu
4408
AGUCAAGGUCUACGCCUAUUACA
4587






AD-1181579.1
uscsaaggUfcUfACfgccuauuacuL96
4230
asdGsuaaUfaggcguadGaCfcuugascsu
4409
AGUCAAGGUCUACGCCUAUUACA
4588






AD-1181580.1
uscsaaggUfcdTadCgccuauuacuL96
4231
asdGsuaaUfaggcguadGaCfcuugascsu
4410
AGUCAAGGUCUACGCCUAUUACA
4589






AD-1181581.1
uscsaaggUfcdTacdGccuauuacuL96
4232
asdGsuaaUfaggcguadGaCfcuugascsu
4411
AGUCAAGGUCUACGCCUAUUACA
4590






AD-569272.6
asasuucuAfcUfAfCfaucuauaacuL96
4233
asGfsuuaUfaGfAfuguaGfuAfgaauususc
4412
GAAAUUCUACUACAUCUAUAACG
4591






AD-1181582.1
asasuucuAfcUfAfCfaucuauaacuL96
4234
asdGsuuaUfagauguaGfuAfgaauususc
4413
GAAAUUCUACUACAUCUAUAACG
4592






AD-1181583.1
asasuucuAfcUfAfCfaucuauaacuL96
4235
asdGsuuaUfagauguadGuAfgaauususc
4414
GAAAUUCUACUACAUCUAUAACG
4593






AD-1181584.1
asasuucuAfcUfAfCfaucuauaacuL96
4236
asdGsuuaUfagaugdTadGudAgaauususc
4415
GAAAUUCUACUACAUCUAUAACG
4594






AD-1181585.1
asasuucuAfcUfAfCfaucuauaacuL96
4237
asdGsuuaUfagauguadGuAfgaauusgsg
4416
GAAAUUCUACUACAUCUAUAACG
4595






AD-1181586.1
asasuucuAfcUfAfCfaucuauaacuL96
4238
asdGsuuaUfagaugdTadGudAgaauusgsg
4417
GAAAUUCUACUACAUCUAUAACG
4596






AD-1181587.1
asasuucuAfcUfAfCfaucuauaacuL96
4239
asdGsuuaUfagauguadGuAfgaasusu
4418
AAUUCUACUACAUCUAUAACG
4597






AD-1181588.1
asasuucuAfcUfAfCfaucuauaacuL96
4240
asdGsuuaUfagaugdTadGudAgaasusu
4419
AAUUCUACUACAUCUAUAACG
4598






AD-1181589.1
asasuucudAcUfAfCfaucuauaacuL96
4241
asGfsuuaUfaGfAfuguaGfuAfgaauususc
4420
AAUUCUACUACAUCUAUAACG
4599






AD-1181590.1
asasuucudAcUfAfCfaucuauaacuL96
4242
asdGsuuaUfagauguadGuAfgaauusgsg
4421
AAUUCUACUACAUCUAUAACG
4600






AD-1181591.1
asasuucudAcUfAfCfaucuauaacuL96
4243
asdGsuuaUfagaugdTadGudAgaauusgsg
4422
AAUUCUACUACAUCUAUAACG
4601






AD-1181592.1
asasuucudAcUfAfCfaucuauaacuL96
4244
asdGsuuaUfagauguadGuAfgaasusu
4423
AAUUCUACUACAUCUAUAACG
4602






AD-1181593.1
asasuucudAcUfAfCfaucuauaacuL96
4245
asdGsuuaUfagaugdTadGudAgaasusu
4424
AAUUCUACUACAUCUAUAACG
4603






AD-565034.2
csasgagaAfaUfUfCfuacuacaucuL96
4246
asGfsaugu(Agn)guagaaUfuUfcucugsusa
4425
UACAGAGAAAUUCUACUACAUCU
4604






AD-1181594.1
csasgagadAaUfUfCfuacuacaucuL96
4247
asdGsaudGu(Agn)guagaaUfuUfcucugsusc
4426
UACAGAGAAAUUCUACUACAUCU
4605






AD-1181595.1
csasgagadAaUfUfCfuacuacaucuL96
4248
asdGsaudGu(A2p)guagaaUfuUfcucugsusc
4427
UACAGAGAAAUUCUACUACAUCU
4606






AD-565035.2
asgsagaaAfuUfCfUfacuacaucuuL96
4249
asAfsgaug(Tgn)aguagaAfuUfucucusgsu
4428
ACAGAGAAAUUCUACUACAUCUA
4607






AD-1181596.1
asgsagaadAuUfCfUfacuacaucuuL96
4250
asAfsgadTg(Tgn)aguagaAfuUfucucusgsu
4429
ACAGAGAAAUUCUACUACAUCUA
4608






AD-1181597.1
asgsagaadAuUfCfUfacuacaucuuL96
4251
asAfsgadTg(U2p)aguagaAfuUfucucusgsu
4430
ACAGAGAAAUUCUACUACAUCUA
4609






AD-1181598.1
asgsagaaAfuUfCfUfacuauaucuuL96
4252
asAfsgadTadTaguagaAfuUfucucusgsu
4431
ACAGAGAAAUUCUACUACAUCUA
4610






AD-565037.2
asgsaaauUfcUfAfCfuacaucuauuL96
4253
asAfsuaga(Tgn)guaguaGfaAfuuucuscsu
4432
AGAGAAAUUCUACUACAUCUAUA
4611






AD-1181599.1
asgsaaauUfcUfAfCfuacaucuauuL96
4254
asAfsuadGa(Tgn)guagdTadGaAfuuucuscsu
4433
AGAGAAAUUCUACUACAUCUAUA
4612






AD-1181600.1
asgsaaauUfcUfAfCfuacaucuauuL96
4255
asAfsuadGa(U2p)guagdTadGaAfuuucuscsu
4434
AGAGAAAUUCUACUACAUCUAUA
4613






AD-1181601.1
asgsaaauUfcUfAfCfuacaucuauuL96
4256
asAfsuadGadT guagdTadGaAfuuucuscsu
4435
AGAGAAAUUCUACUACAUCUAUA
4614






AD-567072.2
csasagguCfuUfCfUfcucuggcuguL96
4257
asCfsagcc(Agn)gagagaAfgAfccuugsasc
4436
GUCAAGGUCUUCUCUCUGGCUGU
4615






AD-1181602.1
csasagguCfuUfCfUfcucuggcuguL96
4258
asCfsagdCc(Agn)gagagaAfgAfccuugsgsc
4437
GUCAAGGUCUUCUCUCUGGCUGU
4616






AD-1181603.1
csasagguCfuUfCfUfcucuggcuguL96
4259
asCfsagdCc(A2p)gagagaAfgAfccuugsgsc
4438
GUCAAGGUCUUCUCUCUGGCUGU
4617






AD-1181604.1
csasagguCfuUfCfUfcucuggcuguL96
4260
asCfsagdCcdAgagagaAfgAfccuugsgsc
4439
GUCAAGGUCUUCUCUCUGGCUGU
4618






AD-567300.2
asgsgagaCfuUfCfCfuugaagccauL96
4261
asUfsggcu(Tgn)caaggaAfgUfcuccusgsc
4440
GCAGGAGACUUCCUUGAAGCCAA
4619






AD-1181605.1
asgsgagaCfuUfCfCfuugaagccauL96
4262
asUfsggdCu(Tgn)caaggaAfgUfcuccusgsc
4441
GCAGGAGACUUCCUUGAAGCCAA
4620






AD-1181606.1
asgsgagaCfuUfCfCfuugaagccauL96
4263
asUfsggdCu(U2p)caaggaAfgUfcuccusgsc
4442
GCAGGAGACUUCCUUGAAGCCAA
4621






AD-567301.2
gsgsagacUfuCfCfUfugaagccaauL96
4264
asUfsuggc(Tgn)ucaaggAfaGfucuccsusg
4443
CAGGAGACUUCCUUGAAGCCAAC
4622






AD-1181607.1
gsgsagacUfuCfCfUfugaagccaauL96
4265
asUfsugdGc(Tgn)ucaadGgAfadGucuccsusg
4444
CAGGAGACUUCCUUGAAGCCAAC
4623






AD-1181608.1
gsgsagacUfuCfCfUfugaagccaauL96
4266
asUfsugdGc(U2p)ucaadGgAfadGucuccsusg
4445
CAGGAGACUUCCUUGAAGCCAAC
4624






AD-569262.2
cscsuacaGfaGfAfAfauucuacuauL96
4267
asUfsaguAfgAfAfuuucUfcUfguaggscsu
4446
AGCCUACAGAGAAAUUCUACUAC
4625






AD-1181609.1
cscsuacadGagAfAfauucuacuauL96
4268
asUfsagdTadGaauuucUfcUfguaggscsu
4447
AGCCUACAGAGAAAUUCUACUAC
4626






AD-1181610.1
cscsuacadGagAfAfauucuacuauL96
4269
asUfsagdTa(G2p)aauuucUfcUfguaggscsu
4448
AGCCUACAGAGAAAUUCUACUAC
4627






AD-569265.2
ascsagagAfaAfUfUfcuacuacauuL96
4270
asAfsuguAfgUfAfgaauUfuCfucugusasg
4449
CUACAGAGAAAUUCUACUACAUC
4628






AD-1181611.1
ascsagagAfaAfUfUfcuacuacauuL96
4271
asAfsugdTadGuagaauUfuCfucugusgsg
4450
CUACAGAGAAAUUCUACUACAUC
4629






AD-1181612.1
ascsagagdAaaUfUfcuacuacauuL96
4272
asdAsugdTadGuagaauUfuCfucugusgsg
4451
CUACAGAGAAAUUCUACUACAUC
4630






AD-569268.2
gsasgaaaUfuCfUfAfcuacaucuauL96
4273
asUfsagaUfgUfAfguagAfaUfuucucsusg
4452
CAGAGAAAUUCUACUACAUCUAU
4631






AD-1181613.1
gsasgaaaUfuCfUfAfcuacaucuauL96
4274
asUfsagaUfguaguagAfaUfuucucsusg
4453
CAGAGAAAUUCUACUACAUCUAU
4632






AD-1181614.1
gsasgaaaUfuCfUfdAcuacaucuauL96
4275
asUfsagdAudGuagudAgdAaUfuucucsusg
4454
CAGAGAAAUUCUACUACAUCUAU
4633






AD-569269.2
asgsaaauUfcUfAfCfuacaucuauuL96
4276
asAfsuagAfuGfUfaguaGfaAfuuucuscsu
4455
AGAGAAAUUCUACUACAUCUAUA
4634






AD-1181615.1
asgsaaauUfcUfAfCfuacaucuauuL96
4277
asAfsuagAfuguagdTadGaAfuuucuscsu
4456
AGAGAAAUUCUACUACAUCUAUA
4635






AD-1181616.1
asgsaaauUfcUfaCfuacaucuauuL96
4278
asdAsuadGadTguagdTadGadAuuucuscsu
4457
AGAGAAAUUCUACUACAUCUAUA
4636






AD-569270.2
gsasaauuCfuAfCfUfacaucuauauL96
4279
asUfsauaGfaUfGfuaguAfgAfauuucsusc
4458
GAGAAAUUCUACUACAUCUAUAA
4637






AD-1181617.1
gsasaauuCfuAfCfUfacaucuauauL96
4280
asUfsaudAgdAuguaguAfgAfauuucsusc
4459
GAGAAAUUCUACUACAUCUAUAA
4638






AD-1181618.1
gsasaauuCfuaCfUfacaucuauauL96
4281
asUfsaudAgdAuguadGudAgdAauuucsusc
4460
GAGAAAUUCUACUACAUCUAUAA
4639






AD-570676.2
ascsccuaCfuCfUfGfuuguucgaauL96
4282
asUfsucgAfaCfAfacagAfgUfagggusasg
4461
CUACCCUACUCUGUUGUUCGAAA
4640






AD-1181619.1
ascsccuaCfuCfUfdGuuguucgaauL96
4283
asUfsucdGadAcaacagAfgUfagggusgsg
4462
CUACCCUACUCUGUUGUUCGAAA
4641






AD-1181620.1
ascsccuaCfuCfUfdGuuguucgaauL96
4284
asUfsucdGa(Agn)caacagAfgUfagggusgsg
4463
CUACCCUACUCUGUUGUUCGAAA
4642






AD-571304.2
csasagguCfuUfCfUfcucuggcuguL96
4285
asCfsagcCfaGfAfgagaAfgAfccuugsasc
4464
GUCAAGGUCUUCUCUCUGGCUGU
4643






AD-1181604.2
csasagguCfuUfCfUfcucuggcuguL96
4286
asCfsagdCcdAgagagaAfgAfccuugsgsc
4465
GUCAAGGUCUUCUCUCUGGCUGU
4644






AD-1181621.1
csasagguCfuUfCfUfcucuggcuguL96
4287
asCfsagdCc(Agn)gagadGadAgdAccuugsgsc
4466
GUCAAGGUCUUCUCUCUGGCUGU
4645






AD-1069946.2
usgsgcucAfaUfGfAfacagagauauL96
4288
asUfsaucu(C2p)uguucaUfuGfagccasasc
4467
GUUGGCUCAAUGAACAGAGAUAC
4646






AD-1181622.1
usgsgcucAfaUfgAfacagagauauL96
4289
asUfsaudCu(C2p)uguudCaUfudGagccasgsc
4468
GUUGGCUCAAUGAACAGAGAUAC
4647






AD-1181623.1
usgsgcucdAaUfgdAacagagauauL96
4290
asUfsaudCu(C2p)uguudCaUfudGagccasgsc
4469
GUUGGCUCAAUGAACAGAGAUAC
4648






AD-1181624.1
usgsgcucdAaUfgdAacagagauauL96
4291
asUfsaudCu(Cgn)uguudCaUfudGagccasgsc
4470
GUUGGCUCAAUGAACAGAGAUAC
4649






AD-1069956.2
gscsugagGfaGfAfAfuugcuucauuL96
4292
asAfsugaa(G2p)caauucUfcCfucagcsasc
4471
GUGCUGAGGAGAAUUGCUUCAUA
4650






AD-1181625.1
gscsugagdGagAfAfuugcuucauuL96
4293
asAfsugdAa(G2p)caauucUfcCfucagcsgsc
4472
GUGCUGAGGAGAAUUGCUUCAUA
4651






AD-1181626.1
gscsugagdGadGadAuugcuucauuL96
4294
asdAsugdAa(G2p)caauucUfcCfucagcsgsc
4473
GUGCUGAGGAGAAUUGCUUCAUA
4652
















TABLE 32







C3 Free Uptake Single Dose Screens


in PCH cells (% C3 mRNA Remaining)
















500 nM

100 nm

10 nM

1 nM




Dose

Dose

Dose

Dose



Duplex
Avg
SD
Avg
SD
Avg
SD
Avg
SD


















AD-564742.5
55
5
50
6
70
7
87
30


AD-1181478.1
51
6
37
6
81
15
95
25


AD-1181479.1
54
4
48
10
77
12
127
4


AD-1181480.1
40
0
53
2
88
0
107
14


AD-1181481.1
54
5
50
10
89
2
143
36


AD-1181482.1
48
19
46
10
81
18
151
58


AD-1181483.1
97
4
82
19
113
33
175
36


AD-1181484.1
52
9
55
3
88
13
97
6


AD-567304.4
51
9
70
5
82
12
89
19


AD-1181485.1
58
4
56
13
79
8
74
8


AD-1181486.1
56
7
46
5
62
9
97
11


AD-1181487.1
55
5
56
10
81
8
95
19


AD-1181488.1
45
1
47
2
72
6
97
4


AD-1181489.1
67
5
81
9
89
9
118
16


AD-1181490.1
95
2
103
13
123
7
138
17


AD-1181491.1
71
8
95
21
111
11
170
28


AD-1181492.1
62
13
91
11
92
7
77
5


AD-567315.8
41
13
54
12
77
2
73
18


AD-1181493.1
50
9
64
7
69
2
110
6


AD-1181494.1
24
3
36
4
62
8
90
6


AD-1181495.1
45
6
69
10
80
12
124
17


AD-1181496.1
58
4
83
26
90
3
105
7


AD-1181497.1
60
7
108
9
100
8
140
5


AD-1181498.1
48
5
62
16
86
3
88
23


AD-1181499.1
64
2
73
5
85
4
122
0


AD-1181500.1
51
4
55
4
87
10
91
11


AD-1181501.1
72
7
75
8
98
19
129
7


AD-1181502.1
55
14
67
5
107
13
133
2


AD-568586.5
40
3
64
4
95
18
146
13


AD-1181503.1
49
2
74
2
95
20
139
18


AD-1181504.1
57
12
57
10
85
10
71
14


AD-1181505.1
41
7
49
6
87
3
84
23


AD-1181506.1
36
0
43
2
81
2
73
3


AD-1181507.1
85
13
83
10
124
5
110
12


AD-1181508.1
56
8
59
1
89
13
118
27


AD-1181509.1
47
7
53
1
89
14
112
15


AD-1181510.1
55
6
56
8
92
12
135
7


AD-568978.5
144
90
77
2
110
12
158
25


AD-1181511.1
72
9
87
15
93
8
79
8


AD-1181513.1
46
2
60
0
100
5
84
2


AD-1181514.1
44
3
52
7
125
18
93
1


AD-1181515.1
86
10
75
4
123
6
135
10


AD-1181516.1
65
3
83
8
92
10
110
18


AD-1181517.1
61
7
81
10
103
10
131
27


AD-569164.9
48
5
44
3
64
10
80
4


AD-1181518.1
39
4
44
5
61
4
83
13


AD-1181519.1
24
3
30
1
70
3
106
4


AD-1181520.1
24
2
28
2
79
10
112
15


AD-1181521.1
51
4
64
7
119
18
134
11


AD-1181522.1
75
6
76
6
117
7
114
5


AD-1181523.1
58
9
60
12
81
8
122
11


AD-1181524.1
50
5
53
10
80
6
108
1


AD-570712.3
37
3
37
4
58
4
71
3


AD-1181525.1
34
9
44
5
70
7
81
2


AD-1181526.1
35
5
47
8
70
2
81
4


AD-1181527.1
35
7
29
4
88
15
106
21


AD-1181528.1
29
0
30
3
99
19
136
8


AD-1181529.1
34
2
36
6
73
7
98
4


AD-1181530.1
34
3
39
13
64
16
84
8


AD-1181531.1
75
10
91
35
110
17
107
26


AD-1181532.1
66
1
77
5
73
16
92
19


AD-1181533.1
61
5
78
8
81
9
81
13


AD-570713.3
48
8
52
11
98
22
110
28


AD-1181534.1
44
6
52
9
96
27
105
15


AD-1181535.1
82
12
74
10
97
5
112
9


AD-1181536.1
75
11
94
26
108
15
133
9


AD-1181537.1
48
4
69
11
89
12
105
4


AD-1181538.1
45
10
70
6
65
12
89
2


AD-1181539.1
45
6
73
5
71
3
74
13


AD-1181540.1
90
4
86
6
84
8
89
9


AD-1181541.1
78
9
116
20
111
24
101
19


AD-1181542.1
86
19
104
15
96
8
108
2


AD-570714.4
37
10
50
9
58
8
94
16


AD-1181543.1
37
5
57
14
61
15
87
20


AD-1181544.1
51
1
60
18
93
16
114
12


AD-1181545.1
62
6
65
9
67
9
71
12


AD-1181546.1
55
6
73
13
85
9
87
7


AD-1181547.1
95
11
88
2
82
1
103
35


AD-1181548.1
89
6
86
8
91
18
103
16


AD-1181549.1
83
13
104
43
87
3
147
62


AD-571826.5
40
3
55
13
76
12
145
36


AD-1181550.1
74
24
78
12
71
6
230
26


AD-1181551.1
70
7
73
7
84
15
68
2


AD-1181552.1
33
6
51
9
80
18
96
20


AD-1181553.1
34
5
50
12
82
19
97
12


AD-1181554.1
68
10
80
17
91
12
100
20


AD-1181555.1
69
17
79
16
109
4
110
21


AD-572040.6
32
4
60
8
91
15
115
19


AD-1181556.1
74
8
121
31
115
19
129
34


AD-1181557.1
64
8
56
5
106
20
89
12


AD-1181558.1
103
6
104
4
150
16
135
14


AD-1181559.1
64
3
86
4
138
21
167
37


AD-1181560.1
99
2
121
26
154
14
155
10


AD-1181561.1
78
7
104
33
137
2
191
35


AD-1181562.1
94
7
81
30
105
20
176
53


AD-1181560.2
122
34
104
9
177
46
161
40


AD-572110.5
41
9
44
7
72
5
139
24


AD-1181563.1
24
5
44
4
97
18
107
11


AD-1181564.1
57
7
50
17
93
14
107
11


AD-1181565.1
42
10
47
7
88
6
133
7


AD-1181566.1
56
6
64
4
95
13
113
17


AD-1181567.1
30
3
46
11
85
5
146
23


AD-1181568.1
88
8
77
2
92
5
148
15


AD-1181569.1
58
12
72
22
134
18
156
5


AD-1181570.1
62
3
83
29
153
50
147
32


AD-1181571.1
25
1
64
8
120
24
86
9


AD-1181572.1
92
10
77
12
114
22
105
12


AD-572387.6
93
18
78
9
91
8
136
33


AD-1181573.1
82
8
91
20
99
7
119
14


AD-1181574.1
102
4
106
5
140
36
132
17


AD-1181575.1
119
12
111
26
100
11
157
17


AD-1181576.1
112
7
124
38
116
8
145
16


AD-1181577.1
91
10
111
16
107
9
95
3


AD-1181578.1
94
13
94
13
109
5
97
8


AD-1181579.1
102
1
97
22
90
10
113
8


AD-1181580.1
102
12
102
16
101
10
122
9


AD-1181581.1
102
13
80
12
91
7
143
9


AD-569272.6
68
10
45
2
65
7
121
17


AD-1181582.1
115
38
97
13
94
16
153
60


AD-1181583.1
82
3
83
8
106
21
90
17


AD-1181584.1
88
2
78
5
93
5
94
24


AD-1181585.1
97
7
94
26
97
5
115
22


AD-1181586.1
98
5
99
21
92
8
128
22


AD-1181587.1
93
14
101
22
90
6
159
49


AD-1181588.1
97
7
89
16
86
24
120
21


AD-1181589.1
52
7
54
15
71
13
134
19


AD-1181590.1
86
7
110
13
101
30
192
56


AD-1181591.1
86
8
87
18
104
6
83
11


AD-1181592.1
89
5
78
7
101
11
87
15


AD-1181593.1
91
3
89
12
97
8
121
32


AD-565034.2
41
1
41
1
63
5
92
5


AD-1181594.1
29
1
36
3
53
3
97
1


AD-1181595.1
25
4
29
4
55
19
100
25


AD-565035.2
30
2
48
17
58
16
127
21


AD-1181596.1
50
2
47
6
88
15
77
10


AD-1181597.1
26
5
29
3
56
3
85
10


AD-1181598.1
61
10
60
1
80
4
103
9


AD-565037.2
35
3
41
1
57
7
133
55


AD-1181599.1
56
7
49
9
60
8
106
13


AD-1181600.1
22
6
23
3
51
20
86
6


AD-1181601.1
20
4
25
9
28
0
84
15


AD-567072.2
64
12
75
6
69
9
112
14


AD-1181602.1
69
4
77
13
115
34
92
11


AD-1181603.1
66
17
55
4
105
18
96
10


AD-1181604.1
54
11
66
16
91
13
106
15


AD-567300.2
51
3
56
4
85
12
137
43


AD-1181605.1
46
3
47
4
62
9
114
22


AD-1181606.1
58
7
65
14
63
15
111
23


AD-567301.2
45
15
41
12
52
2
101
9


AD-1181607.1
29
0
43
2
78
18
128
14


AD-1181608.1
36
6
35
2
95
21
79
9


AD-569262.2
18
2
23
2
45
6
78
9


AD-1181609.1
20
5
21
1
47
12
106
25


AD-1181610.1
19
2
27
1
37
1
140
5


AD-569265.2
41
7
32
5
44
7
108
12


AD-1181611.1
16
3
24
8
38
8
115
34


AD-1181612.1
28
1
37
13
72
29
164
55


AD-569268.2
21
2
24
3
54
13
60
11


AD-1181613.1
32
5
26
4
45
9
60
9


AD-1181614.1
28
6
30
4
44
9
77
10


AD-569269.2
19
4
16
3
25
0
67
9


AD-1181615.1
16
1
18
3
26
0
95
27


AD-1181616.1
36
6
26
3
51
25
105
15


AD-569270.2
43
8
51
13
68
15
105
32


AD-1181617.1
24
1
40
6
59
11
99
24


AD-1181618.1
38
3
39
7
64
8
77
4


AD-570676.2
67
19
39
1
49
4
117
7


AD-1181619.1
36
10
35
5
41
6
87
2


AD-1181620.1
68
10
58
9
57
3
127
14


AD-571304.2
70
1
66
11
60
2
215
100


AD-1181604.2
69
5
63
8
101
39
162
36


AD-1181621.1
108
12
150
48
105
5
188
54


AD-1069946.2
83
15
91
14
149
44
67
8


AD-1181622.1
70
4
85
26
137
21
124
17


AD-1181623.1
70
11
76
22
120
48
120
17


AD-1181624.1
95
12
83
10
119
3
129
22


AD-1069956.2
51
9
57
15
102
23
130
16


AD-1181625.1
55
1
46
13
80
27
138
29


AD-1181626.1
60
7
48
10
98
20
128
31
















TABLE 33







C3 Transfection Single Dose Screens


in PCH cells (% C3 mRNA Remaining)
















50 nM

10 nM

1 nM

0.1 nm




Dose

Dose

Dose

Dose



Duplex
Avg
SD
Avg
SD
Avg
SD
Avg
SD


















AD-564742.5
4
0
6
4
54
2
64
7


AD-1181478.1
4
0
4
3
134
8
55
3


AD-1181479.1
5
1
4
3
137
33
73
17


AD-1181480.1
4
0
4
3
137
8
78
15


AD-1181481.1
6
1
4
2
118
21
77
6


AD-1181482.1
5
1
4
2
98
14
38
3


AD-1181483.1
8
2
7
3
107
10
105
17


AD-1181484.1
6
0
4
1
27
5
58
15


AD-567304.4
6
1
7
1
44
12
63
0


AD-1181485.1
3
1
6
0
23
5
60
8


AD-1181486.1
4
1
10
5
45
9
55
10


AD-1181487.1
5
2
9
5
49
7
70
2


AD-1181488.1
4
1
5
1
44
7
64
9


AD-1181489.1
7
2
7
2
103
30
76
2


AD-1181490.1
7
3
6
3
105
12
108
19


AD-1181491.1
8
2
6
1
90
20
81
17


AD-1181492.1
6
2
9
2
75
14
89
14


AD-567315.8
4
1
10
1
76
5
48
7


AD-1181493.1
6
1
7
0
105
24
60
8


AD-1181494.1
5
0
5
0
112
21
44
4


AD-1181495.1
7
1
6
1
129
28
65
18


AD-1181496.1
9
1
5
2
133
31
75
11


AD-1181497.1
6
1
2
3
93
12
61
2


AD-1181498.1
7
1
16
5
120
28
70
12


AD-1181499.1
6
1
15
3
101
20
74
11


AD-1181500.1
8
0
10
2
107
7
72
6


AD-1181501.1
9
1
13
1
109
1
71
8


AD-1181502.1
11
2
14
2
125
19
76
2


AD-568586.5
7
2
5
2
78
26
54
10


AD-1181503.1
7
3
5
0
82
11
36
3


AD-1181504.1
5
1
7
2
81
7
39
10


AD-1181505.1
5
1
8
2
77
6
37
1


AD-1181506.1
5
0
9
1
86
10
41
3


AD-1181507.1
22
2
29
3
89
1
90
9


AD-1181508.1
9
2
10
2
94
5
62
3


AD-1181509.1
6
0
7
1
97
4
55
4


AD-1181510.1
7
2
6
1
49
8
46
5


AD-568978.5
8
0
9
3
101
18
74
23


AD-1181511.1
8
1
14
5
67
11
69
11


AD-1181513.1
6
0
11
3
81
7
59
5


AD-1181514.1
6
0
7
1
24
4
55
9


AD-1181515.1
65
8
62
1
25
4
99
5


AD-1181516.1
37
2
31
3
17
5
94
14


AD-1181517.1
41
5
38
5
19
3
88
12


AD-569164.9
4
1
7
1
19
1
36
4


AD-1181518.1
6
1
7
1
12
1
46
6


AD-1181519.1
6
1
7
1
33
6
48
6


AD-1181520.1
6
0
7
1
25
2
53
5


AD-1181521.1
12
1
15
1
24
4
74
3


AD-1181522.1
32
3
31
1
11
2
83
6


AD-1181523.1
14
1
9
1
14
4
68
5


AD-1181524.1
7
1
6
1
78
2
51
5


AD-570712.3
5
1
6
0
50
9
47
2


AD-1181525.1
5
0
6
1
43
4
62
12


AD-1181526.1
7
0
5
1
50
11
63
10


AD-1181527.1
6
1
6
1
35
4
54
9


AD-1181528.1
5
1
5
1
19
5
51
3


AD-1181529.1
6
1
5
1
23
4
62
12


AD-1181530.1
5
1
5
0
18
5
40
5


AD-1181531.1
22
1
18
5
13
1
89
7


AD-1181532.1
12
1
10
3
16
3
74
17


AD-1181533.1
15
1
17
1
13
2
76
7


AD-570713.3
6
0
6
1
16
2
56
6


AD-1181534.1
8
1
7
1
21
4
66
8


AD-1181535.1
23
4
16
1
19
4
104
20


AD-1181536.1
25
5
16
2
10
1
113
57


AD-1181537.1
8
2
5
1
15
1
77
50


AD-1181538.1
2
1
5
1
8
2
44
5


AD-1181539.1
4
0
8
2
10
0
58
2


AD-1181540.1
30
2
40
3
13
3
93
3


AD-1181541.1
122
20
89
5
10
1
99
12


AD-1181542.1
88
12
79
12
8
0
97
13


AD-570714.4
6
1
5
0
11
6
47
5


AD-1181543.1
4
1
4
0
19
1
41
6


AD-1181544.1
6
1
10
2
15
2
66
10


AD-1181545.1
5
1
10
1
16
2
74
3


AD-1181546.1
9
0
17
4
17
5
79
7


AD-1181547.1
77
15
62
4
14
0
85
7


AD-1181548.1
66
9
65
7
40
4
87
2


AD-1181549.1
35
2
49
19
91
25
87
6


AD-571826.5
5
0
6
1
39
1
55
2


AD-1181550.1
8
1
8
0
72
9
70
13


AD-1181551.1
7
2
7
1
36
3
79
4


AD-1181552.1
4
0
5
2
37
10
59
2


AD-1181553.1
3
2
7
1
40
14
50
6


AD-1181554.1
27
9
25
6
68
10
89
2


AD-1181555.1
24
10
17
1
25
7
120
34


AD-572040.6
5
2
8
3
26
15
72
10


AD-1181556.1
22
4
42
1
20
1
122
22


AD-1181557.1
5
0
17
4
54
2
97
9


AD-1181558.1
40
3
79
10
134
8
92
14


AD-1181559.1
16
5
59
2
137
33
113
17


AD-1181560.1
45
28
77
3
137
8
102
10


AD-1181561.1
22
12
60
4
118
21
91
5


AD-1181562.1
119
96
63
1
98
14
99
10


AD-1181560.2
113
110
71
11
107
10
117
2


AD-572110.5
3
0
10
2
27
5
106
21


AD-1181563.1
4
0
14
3
44
12
94
18


AD-1181564.1
5
0
9
1
23
5
54
17


AD-1181565.1
6
1
13
3
45
9
84
3


AD-1181566.1
7
1
14
2
49
7
84
3


AD-1181567.1
5
1
16
4
44
7
89
18


AD-1181568.1
21
4
35
0
103
30
108
14


AD-1181569.1
21
8
52
7
105
12
108
16


AD-1181570.1
14
4
36
3
90
20
104
7


AD-1181571.1
9
4
22
1
75
14
95
12


AD-1181572.1
18
1
23
1
76
5
83
12


AD-572387.6
26
3
53
7
105
24
118
9


AD-1181573.1
44
10
64
2
112
21
112
27


AD-1181574.1
66
15
80
1
129
28
141
25


AD-1181575.1
91
9
89
12
133
31
132
20


AD-1181576.1
71
22
99
9
93
12
104
7


AD-1181577.1
88
21
94
12
120
28
83
6


AD-1181578.1
88
9
95
13
101
20
93
12


AD-1181579.1
99
17
102
13
107
7
102
16


AD-1181580.1
88
3
100
1
109
1
122
8


AD-1181581.1
87
11
104
12
125
19
115
16


AD-569272.6
8
0
17
2
78
26
110
3


AD-1181582.1
22
4
33
4
82
11
99
6


AD-1181583.1
57
8
80
3
81
7
76
11


AD-1181584.1
41
3
64
10
77
6
100
20


AD-1181585.1
69
3
82
9
86
10
93
11


AD-1181586.1
67
4
75
13
89
1
136
3


AD-1181587.1
65
6
83
5
94
5
117
24


AD-1181588.1
63
6
80
8
97
4
117
4


AD-1181589.1
8
2
16
5
49
8
110
21


AD-1181590.1
68
13
62
6
101
18
106
10


AD-1181591.1
60
5
78
4
67
11
85
8


AD-1181592.1
83
2
89
8
83
4
98
8


AD-1181593.1
80
1
89
8
81
7
99
4


AD-565034.2
7
1
9
2
24
4
82
11


AD-1181594.1
5
1
8
1
25
4
73
15


AD-1181595.1
5
1
8
1
17
5
69
11


AD-565035.2
5
1
10
1
19
3
69
28


AD-1181596.1
6
1
9
1
19
1
40
11


AD-1181597.1
5
1
8
1
12
1
42
6


AD-1181598.1
9
0
14
0
33
6
92
11


AD-565037.2
5
1
7
1
25
2
62
3


AD-1181599.1
6
0
9
0
24
4
70
9


AD-1181600.1
4
1
6
1
11
2
50
15


AD-1181601.1
4
0
6
2
14
4
32
6


AD-567072.2
20
2
40
4
78
2
119
6


AD-1181602.1
11
3
22
2
50
9
85
22


AD-1181603.1
10
1
19
5
43
4
81
5


AD-1181604.1
12
0
19
2
50
11
100
9


AD-567300.2
7
1
14
3
35
4
102
14


AD-1181605.1
4
1
7
1
19
5
62
1


AD-1181606.1
5
1
11
2
23
4
105
5


AD-567301.2
4
0
6
0
18
5
72
12


AD-1181607.1
4
0
6
1
13
1
82
46


AD-1181608.1
4
1
8
0
16
3
32
5


AD-569262.2
4
1
7
1
13
2
41
5


AD-1181609.1
4
1
9
2
16
2
51
3


AD-1181610.1
4
0
7
1
21
4
71
9


AD-569265.2
5
1
9
1
19
4
54
12


AD-1181611.1
3
1
5
1
10
1
55
21


AD-1181612.1
5
0
7
1
15
1
51
9


AD-569268.2
3
1
5
1
8
2
18
2


AD-1181613.1
4
0
6
0
10
0
25
5


AD-1181614.1
4
0
7
1
13
3
37
4


AD-569269.2
3
0
5
1
10
1
31
8


AD-1181615.1
3
1
6
0
8
0
28
7


AD-1181616.1
5
0
8
1
11
6
34
4


AD-569270.2
3
1
9
2
19
1
74
16


AD-1181617.1
4
0
5
1
15
2
34
10


AD-1181618.1
6
1
8
1
16
2
37
3


AD-570676.2
6
0
8
1
17
5
36
9


AD-1181619.1
6
0
7
1
14
0
38
5


AD-1181620.1
8
1
14
4
40
4
75
10


AD-571304.2
16
5
25
8
91
25
130
30


AD-1181604.2
10
2
18
4
39
1
107
22


AD-1181621.1
26
4
89
30
72
9
86
37


AD-1069946.2
9
0
19
9
36
3
61
14


AD-1181622.1
5
2
7
3
37
10
58
10


AD-1181623.1
9
2
11
4
40
14
81
16


AD-1181624.1
16
2
17
3
68
10
91
14


AD-1069956.2
6
0
6
1
25
7
70
9


AD-1181625.1
5
0
6
3
26
15
59
7


AD-1181626.1
5
1
10
1
20
1
60
10









Example 6. In Vivo Screening of dsRNA Duplexes in Mice

Duplexes of interest, identified from the above in vitro studies were evaluated in vivo. In particular, at pre-dose day—14 groups of wild-type mice (C57BL/6) (n=3) were transduced by intravenous administration of 2×1010 viral particles of an adeno-associated virus 8 (AAV8) vector encoding human complement component C3. In particular, mice were administered an AAV8 encoding a portion of human complement component C3 mRNA spanning nucleotides 94-2892 of NM_000064.3.


At day 0, groups of three mice were subcutaneously administered a single 10 mg/kg dose of the agents of interest or PBS control. At day 14 post-dose animals were sacrificed, liver samples were collected and snap-frozen in liquid nitrogen. Tissue mRNA was extracted and analyzed by the RT-QPCR method.


Human C3 mRNA levels were compared to housekeeping gene GAPDH. The values were then normalized to the average of PBS vehicle control group. The data were expressed as percent of baseline value, and presented as mean plus standard deviation. The results, presented in FIG. 4, demonstrate that the exemplary duplex agents tested effectively reduce the level of the human C3 messenger RNA in vivo.


Example 7. In Vivo Analysis of Duplexes of Interest in Non-Human Primates

Duplexes of interest, identified from the above in vitro and in vivo studies, were evaluated in vivo. In particular, female Cynomolgus monkeys were subcutaneously administered a single dose of the agents of interest. FIG. 5 provides the treatment groups and the duplexes of interest. Serum was collected weekly to the end of the study and C3 protein levels were determined by ELISA assay (C3 Human ELISA: Hycult HK366). Briefly, this human C3 ELISA assay was previously validated for cross reactivity to cynomolgus monkey and the instructions provided with the kit were followed with the exceptions that samples were diluted 1:50,000 or 1:20,000 for samples with high silencing expected in order to keep ODs within the standard curve. ELISA assays were performed at interim time points, and any data that was reproduced twice was averaged at the g/ml level then normalized to average pre-dose.


The results, shown in FIG. 6, demonstrate that the exemplary duplex agents tested potently and durably reduce the level of the Cynomolgus C3 protein in vivo.


Additional duplexes of interest, identified from the above in vitro and in vivo studies, were also evaluated in vivo. In particular, nine Groups of female Cynomolgus monkeys were subcutaneously administered a single dose of the agents of interest (Groups 1-5 and 7-10) and one Group of emale Cynomolgus monkeys was subcutaneously administered two doses of a duplex of interest on Day 1 and Day 55. FIG. 7 provides the treatment groups and the duplexes of interest. Serum was collected weekly to the end of the study and C3 protein levels were determined by ELISA assay (C3 Human ELISA: Hycult HK366). Briefly, this human C3 ELISA assay was previously validated for cross reactivity to cynomolgus monkey and the instructions provided with the kit were followed with the exceptions that samples were diluted 1:50,000 or 1:15,000 for samples with high silencing expected in order to keep ODs within the standard curve. ELISA assays were performed at interim time points, and any data that was reproduced twice was averaged at the g/ml level then normalized to average pre-dose.


The results, shown in FIG. 8, demonstrate that the exemplary duplex agents tested potently and durably reduce the level of the Cynomolgus C3 protein in vivo.


In another study, additional duplexes of interest, identified from the above in vitro and in vivo studies, were evaluated in vivo. In particular, three Groups of female Cynomolgus monkeys were subcutaneously administered a single 3 mg/kg dose of AD-1181519, AD-569268, or AD-570714 (Groups 1, 4, and 7), three Groups of female Cynomolgus monkeys were subcutaneously administered a single 9 mg/kg dose of AD-1181519, AD-569268, or AD-570714 (Groups 2, 5, and 8), one Group of female Cynomolgus monkeys was subcutaneously administered a single 25 mg/kg dose of AD-570714 (Group 10), and three Groups of female Cynomolgus monkeys were subcutaneously administered three 3 mg/kg dose of AD-1181519, AD-569268, or AD-570714 on Days 1, 29, and 57 (Groups 3, 6, and 9; 3×3 mg/kg) (see FIG. 8). Serum was collected weekly to the end of the study. C3 protein levels were determined by ELISA assay (C3 Human ELISA: Hycult HK366) and hemolytic activity was evaluated to determine the functional activity of the alternative pathway, e.g., alternative hemolysis assay Wieslsab Complement Alternative Pathway (CAP) assay. For the C3 ELISA assays, the assay used was previously validated for cross reactivity to cynomolgus monkey and the instructions provided with the kit were followed with the exceptions that samples were diluted 1:39,067. ELISA assays were performed at interim time points, and any data that was reproduced twice was averaged at the μg/ml level then normalized to average pre-dose. In addition, liver biopsies were performed on 3 animals administered 25 mg/kg of AD-570714 at Days −21 and Day 29 (see FIG. 9).


The results, shown in FIG. 10, demonstrate that the exemplary duplex agents tested potently and durably reduce the level of the Cynomolgus C3 protein in vivo.


EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments and methods described herein. Such equivalents are intended to be encompassed by the scope of the following claims.

Claims
  • 1. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C3 in a cell, or a pharmaceutically acceptable salt thereof, comprising a sense strand differing by no more than 4 bases from the nucleotide sequence 5′-gsasgccgUfuCfUfCfuacaauuacu-3′ of SEQ ID NO:4188 and an antisense strand differing by no more than 4 bases from the nucleotide sequence 5′-asGfsuaaUfuGfUfagagAfaCfggcucsgsg-3′ of SEQ ID NO:4367, wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C and U, respectively; and s is a phosphorothioate linkage.
  • 2. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, comprising a sense strand differing by no more than 3 bases from the nucleotide sequence 5′-gsasgccgUfuCfUfCfuacaauuacu-3′ of SEQ ID NO:4188 and an antisense strand differing by no more than 3 bases from the nucleotide sequence 5′-asGfsuaaUfuGfUfagagAfaCfggcucsgsg-3′ of SEQ ID NO:4367.
  • 3. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, comprising a sense strand differing by no more than 2 bases from the nucleotide sequence 5′-gsasgccgUfuCfUfCfuacaauuacu-3′ of SEQ ID NO:4188 and an antisense strand differing by no more than 2 bases from the nucleotide sequence 5′-asGfsuaaUfuGfUfagagAfaCfggcucsgsg-3′ of SEQ ID NO:4367.
  • 4. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, comprising a sense strand differing by no more than 1 base from the nucleotide sequence 5′-gsasgccgUfuCfUfCfuacaauuacu-3′ of SEQ ID NO:4188 and an antisense strand differing by no more than 1 base from the nucleotide sequence 5′-asGfsuaaUfuGfUfagagAfaCfggcucsgsg-3′ of SEQ ID NO:4367.
  • 5. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, comprising a sense strand comprising the nucleotide sequence 5′-gsasgccgUfuCfUfCfuacaauuacu-3′ of SEQ ID NO:4188 and an antisense strand comprising the nucleotide sequence 5′-asGfsuaaUfuGfUfagagAfaCfggcucsgsg-3′ of SEQ ID NO:4367.
  • 6. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, comprising a sense strand consisting of the nucleotide sequence 5′-gsasgccgUfuCfUfCfuacaauuacu-3′ of SEQ ID NO:4188 and an antisense strand consisting of the nucleotide sequence 5′-asGfsuaaUfuGfUfagagAfaCfggcucsgsg-3′ of SEQ ID NO:4367.
  • 7. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, further comprising a ligand.
  • 8. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 7, wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent.
  • 9. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 7, wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
  • 10. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 9, wherein the ligand is one or more GalNAc derivatives attached through a monovalent, bivalent, or trivalent linker.
  • 11. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 10, wherein the ligand is
  • 12. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 11, wherein the dsRNA agent is conjugated to the ligand as shown in the following schematic
  • 13. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 12, wherein X is O.
  • 14. An isolated cell containing the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1.
  • 15. A pharmaceutical composition comprising the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1.
  • 16. The pharmaceutical composition of claim 15, wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is in an unbuffered solution.
  • 17. The pharmaceutical composition of claim 16, wherein the unbuffered solution is saline or water.
  • 18. The pharmaceutical composition of claim 15, wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is in a buffer solution.
  • 19. The pharmaceutical composition of claim 18, wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof.
  • 20. The pharmaceutical composition of claim 19, wherein the buffer solution is phosphate buffered saline (PBS).
  • 21. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C3 in a cell, or a pharmaceutically acceptable salt thereof, comprising a sense strand comprising the nucleotide sequence 5′-gsasgccgUfuCfUfCfuacaauuacu-3′ of SEQ ID NO:4188 and an antisense strand comprising the nucleotide sequence 5′-asGfsuaaUfuGfUfagagAfaCfggcucsgsg-3′ of SEQ ID NO:4367, wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C and U, respectively; and s is a phosphorothioate linkage,wherein the 3′-end of the sense strand of the dsRNA agent is conjugated to a ligand as shown in the following schematic
  • 22. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 21, which is in a sodium salt form.
  • 23. An isolated cell containing the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 21.
  • 24. A pharmaceutical composition comprising the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 21.
  • 25. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C3 in a cell, or a pharmaceutically acceptable salt thereof, consisting of a sense strand consisting of the nucleotide sequence 5′-gsasgccgUfuCfUfCfuacaauuacu-3′ of SEQ ID NO:4188 and an antisense strand consisting of the nucleotide sequence 5′-asGfsuaaUfuGfUfagagAfaCfggcucsgsg-3′ of SEQ ID NO:4367, wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C and U, respectively; and s is a phosphorothioate linkage,wherein the 3′-end of the sense strand of the dsRNA agent is conjugated to a ligand as shown in the following schematic
  • 26. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 25, which is in a sodium salt form.
  • 27. An isolated cell containing the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 25.
  • 28. A pharmaceutical composition comprising the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 25.
RELATED APPLICATIONS

This application is a continuation of Ser. No. 17/721,530, filed on Apr. 15, 2022, which is a 35 § U.S.C. 111(a) continuation application which claims the benefit of priority to PCT/US2020/056563, filed on Oct. 21, 2020, which claims the benefit of priority to U.S. Provisional Application No. 62/924,210, filed on Oct. 22, 2019. The entire contents of each of the foregoing applications are incorporated herein by reference.

Provisional Applications (1)
Number Date Country
62924210 Oct 2019 US
Continuations (2)
Number Date Country
Parent 17721530 Apr 2022 US
Child 18594132 US
Parent PCT/US2020/056563 Oct 2020 WO
Child 17721530 US