REVERSIR MOLECULES AND METHODS OF USE THEREOF

1 . A single stranded oligonucleotide for inhibiting RNAi activity of a double stranded ribonucleic acid (dsRNA) agent comprising a thermally destabilizing nucleotide in the antisense strand,

wherein the single stranded oligonucleotide comprises a nucleotide sequence substantially complementary to the antisense strand of the dsRNA agent,

wherein the single stranded oligonucleotide is 16-30 nucleotides in length,

wherein substantially all of the nucleotides of the single stranded oligonucleotide comprise a nucleotide modification, and

wherein at least three of the nucleotide modifications are a high affinity nucleotide modification.

2 . The single stranded oligonucleotide of claim 1 ,

(i) wherein the thermally destabilizing nucleotide modification is selected from the group consisting of an abasic modification; a mismatch with the opposing nucleotide in the duplex; a destabilizing sugar modification, a 2′-deoxy modification, an acyclic nucleotide, an unlocked nucleic acid (UNA), and a glycerol nucleic acid (GNA);

(ii) wherein substantially all of the nucleotides comprise a nucleotide modification selected from the group consisting of a 2′-O-alkyl modification, a 2′ substituted alkoxy modification, a 2′-substituted alkyl modification, a 2′-halo modification, a deoxynucleotide modification, a locked nucleic acid (LNA) modification, a D-Methyleneoxy (4′-CH2-O-2′) locked nucleic acid (LNA) modification, a 2-O-(2-Methoxyethyl) (MOE) modification, bridged nucleic acid (2′,4′-BNA), 2-O-Ethyl (cEt), and a 2′-O-methyl modification;

(iii) wherein all of the nucleotides comprise a nucleotide modification selected from the group consisting of a 2′-O-alkyl modification, a 2′-substituted alkoxy modification, a 2′-substituted alkyl modification, a 2′-halo modification, a deoxynucleotide modification, a D-Methyleneoxy (4′-CH2-O-2′) locked nucleic acid (LNA) modification, bridged nucleic acid (2′,4′-BNA), 2′-O-Ethyl (cEt), and a 2′-O-methyl modification;

(iv) wherein at least four of the nucleotide modifications are a high affinity nucleotide modification;

(v) wherein at least five of the nucleotide modifications are a high affinity nucleotide modification; and/or

(vi) further comprising at least five phosphorothioate internucleotide modifications.

3 - 5 . (canceled)

6 . The single stranded oligonucleotide of claim 1 ,

(i) wherein at least two of the high affinity nucleotide modifications are at positions 2 and 6; positions 2 and 5; positions 2 and 7; positions 2 and 8; positions 2 and 9; positions 2 and 14; positions 2 and 15; and/or positions 2 and 16, counting from the 3′-end of the oligonucleotide;

(ii) wherein the high affinity nucleotide modifications are at positions 2, 6, 8, and 14; 2, 4, 5, 6, and 7; 2, 4, 6, 8, and 13; 2, 4, 6, 8, and 14; 2, 4, 6, 8, and 15; 2, 4, 6, 8, and 16; 2, 8, 10, and 14; 2, 4, 6, 8, and 14; or 2, 8, 12, and 14, counting from the 3′-end of the oligonucleotide;

(iii) wherein at least one of the nucleotides comprising a high affinity nucleotide modification is base paired with the nucleotide comprising the thermally destabilizing nucleotide in the antisense strand of the dsRNA agent; and/or

(iv) wherein the high affinity modification is selected from the group consisting of a locked nucleic acid (LNA) modification, a constrained Ethyl nucleic acid (cEtNA) modification, and a bridged nucleic acid (BNA) modification.

7 - 10 . (canceled)

11 . The single stranded oligonucleotide of claim 1 , wherein the single stranded oligonucleotide is conjugated to at least one ligand.

12 . The single stranded oligonucleotide of claim 11 ,

(i) wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative;

(ii) wherein the ligand is one or more GalNAc derivatives attached through a monovalent, bivalent, or trivalent branched linker;

(iii) wherein the ligand is

(iv) wherein the ligand is conjugated to a nucleoside comprising a deoxy sugar in the single stranded oligonucleotide, optionally wherein the deoxy sugar is a 2′-deoxy ribose; and/or

(v) wherein the ligand is conjugated to 3′-terminus of the single stranded oligonucleotide.

13 - 17 . (canceled)

18 . A single stranded oligonucleotide for inhibiting RNAi activity of a double stranded ribonucleic acid (dsRNA) agent comprising a thermally destabilizing nucleotide in the antisense strand,

wherein the single stranded oligonucleotide comprises a nucleotide sequence substantially complementary to the antisense strand of the dsRNA agent,

wherein the single stranded oligonucleotide is 18-24 nucleotides in length,

wherein the single stranded oligonucleotide comprises at least five phosphorothioate internucleotide modifications and is represented by formula (I):

wherein:

B1, B2 and B3 each independently represent a nucleotide comprising a nucleotide modification independently selected from the group consisting of a 2′-deoxy, 2′-ribo, 2′-O-alkyl modification, a 2′-substituted alkoxy modification, a 2′-substituted alkoxy alkyl modification, a 2′-substituted alkyl modification, and a 2′-halo modification;

T1, T2, and T3 each independently represent a nucleotide comprising a nucleotide modification selected from the group consisting of a deoxynucleotide modification, a D-Methyleneoxy (4′-CH2-O-2′) locked nucleic acid (LNA) modification, a 2′O-(2-Methoxyethyl) (MOE) modification, a bridged nucleic acid (2′,4′-BNA) modification, 2′-O-Ethyl (cEt) modification, a 2′-deoxy-2′-Fluoro, and a 2′-O-methyl modification;

q 1 , q 3 and q 5 are each independently 3-12 nucleotides in length;

q 2 , q 4 and q 6 are independently 1-6 nucleotide(s) in length; and

wherein the single stranded oligonucleotide is conjugated to at least one ligand.

19 . The single stranded oligonucleotide of claim 1 ,

(i) wherein the single stranded oligonucleotide comprises 5-15 phosphorothioate internucleotide modifications; 5-14 phosphorothioate internucleotide modifications; 5-13 phosphorothioate internucleotide modifications; 5-12 phosphorothioate internucleotide modifications; 5-11 phosphorothioate internucleotide modifications; 5-10 phosphorothioate internucleotide modifications; 5-9 phosphorothioate internucleotide modifications; 5-8 phosphorothioate internucleotide modifications; 5-7 phosphorothioate internucleotide modifications; or 5-6 phosphorothioate internucleotide modifications: or

(ii) wherein the single stranded oligonucleotide comprises 6-14 phosphorothioate internucleotide modifications.

20 . (canceled)

21 . (canceled)

22 . The single stranded oligonucleotide of claim 1 ,

(i) wherein the single stranded oligonucleotide is at least about 90% complementary to the entire length of the antisense strand of the dsRNA agent;

(ii) wherein the single stranded oligonucleotide is 90% complementary to nucleotides 2-16 of the antisense stand of the dsRNA agent;

(iii) wherein the single stranded oligonucleotide is fully complementary to the antisense strand of the dsRNA agent;

(iv) wherein the nucleotide sequence of the antisense strand of the dsRNA agent comprises the nucleotide sequence 5′-UGUACUCUCAUUGUGGAUGACGA-3′ of SEQ ID NO: 9;

(v) wherein the nucleotide sequence of the antisense strand of the dsRNA agent comprises the nucleotide sequence 5′-usGfsuac(Tgn)cucauugUfgGfaugacsgsa-3′ of SEQ ID NO: 11, wherein a, c, g, and u are 2′-O-methyladenosine-3′-phosphate, 2′-O-methylcytidine-3′-phosphate, 2′-O-methylguanosine-3′-phosphate, and 2′-O-methyluridine-3′-phosphate, respectively; Af, Cf, Gf, and Uf are 2′-O-fluoroadenosine-3′-phosphate, 2′-O-fluorocytidine-3′-phosphate, 2′-O-fluoroguanosine-3′-phosphate, and 2′-O-fluorouridine-3′-phosphate, respectively; dT is a deoxy-thymine; s is a phosphorothioate linkage; and (Tgn) is thymidine-glycol nucleic acid (GNA)S-isomer;

(vi) wherein the nucleotide sequence of the single stranded oligonucleotide is at least 90% identical to the entire nucleotide sequence of any one of the unmodified nucleotide sequences in Table 6; and/or

(vii) wherein the nucleotide sequence of the single stranded oligonucleotide comprises a modified nucleotide sequence differing by no more than 4 modified nucleotides from any one of the modified nucleotide sequences in Table 6.

23 - 28 . (canceled)

29 . The single stranded oligonucleotide of claim 18 ,

(i) wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative;

(ii) wherein the ligand is one or more GalNAc derivatives attached through a monovalent, bivalent, or trivalent branched linker;

(iii) wherein the ligand is

(iv) wherein the ligand is conjugated to a nucleoside comprising a deoxy sugar in the single stranded oligonucleotide, optionally wherein the deoxy sugar is a 2′-deoxy ribose; and/or

(v) wherein the ligand is conjugated to 3′-terminus of the single stranded oligonucleotide.

30 - 35 . (canceled)

36 . A pharmaceutical composition comprising the single stranded oligonucleotide of claim 1 .

37 - 40 . (canceled)

41 . An isolated cell comprising the single stranded oligonucleotide of claim 1 .

42 . A method of inhibiting the RNAi inhibitory activity of a dsRNA agent comprising a thermally destabilizing nucleotide modification in the antisense strand, the method comprising

contacting the dsRNA agent with the single stranded oligonucleotide of claim 1 , thereby inhibiting the RNAi inhibitory activity of a dsRNA agent comprising a thermally destabilizing nucleotide modification in the antisense strand.

43 . (canceled)

44 . (canceled)

45 . A method of treating a subject in need thereof, the method comprising

administering to the subject a therapeutically effective amount of the single stranded oligonucleotide of claim 1 , thereby treating the subject.

46 . The method of claim 45 , wherein the subject in need thereof was previously administered a double stranded RNAi agent that inhibits the expression of a target gene and comprises a thermally destabilizing nucleotide modification in the antisense strand.

47 . The method of claim 46 , wherein the target gene is angiotensinogen (AGT), and/or wherein the subject in need thereof is suffering from hypotension, hyperkalemia, or renal dysfunction.

48 - 50 . (canceled)

51 . The method of claim 45 , further comprising administering to the subject an additional therapy or therapeutic agent selected from the group consisting of increased dietary fluid/salt, fludrocortisone/midodrine treatment, intravenous fluids, vasopressor medications, down-titration or interruption of concomitant antihypertensive medications, a low potassium diet, thiazide/loop diuretic medications, oral potassium binders, calcium, glucose, insulin, and hemodialysis, or combinations thereof.

52 . The method of claim 45 , wherein the single stranded oligonucleotide is administered to the subject subcutaneously or intravenously.

53 . (canceled)

54 . The method of claim 45 , wherein the dose of the single stranded oligonucleotide is at a ratio of about 1:1, 2:1 or 3:1 to the dose of dsRNA agent previously administered to the subject.

55 . A method of ameliorating in a subject a side effect of a dsRNA agent which inhibits the expression of a target gene and comprises a thermally destabilizing nucleotide modification in the antisense strand, the method comprising

administering to the subject an effective amount of the single stranded oligonucleotide of claim 1 , thereby ameliorating the side effect of the dsRNA agent in the subject.

56 - 66 . (canceled)

67 . A kit comprising the single stranded oligonucleotide of claim 1 , and instructions for use, and optionally, means for administration of the single stranded oligonucleotide.

68 . (canceled)