Modified double-stranded rna agents

A double-stranded RNA (dsRNA) comprising a sense strand having 21 nucleotides and an antisense strand having 23 nucleotides that is at least 95% complementary to a portion of a target mRNA of a target gene, wherein:

• (a) each nucleotide of the sense and antisense strands has a modification independently selected from the group consisting of 2′-O-Methyl (2′-OMe) and 2′-deoxy-2′-fluoro (2′-F);
• (b) the sense strand comprises 2′-F modifications only at positions 7, 9, 10, and 11, counting from the 5′-end of the sense strand;
• (c) the antisense strand comprises 2′-F modifications only at positions 2, 6, 9, 14, and 16, counting from the 5′-end of the antisense strand;
• (d) the sense and antisense strands form a duplex region that is 21 base pairs in length;
• (e) the dsRNA is covalently conjugated to a ligand at the 3′-end of the sense strand;
• (f) the dsRNA has a 2 nucleotide overhang at the 3′-end of the antisense strand;
• and
• (g) phosphorothioate internucleotide linkage modifications, when present, only occur within a terminal region at one or both termini of the sense strand, one or both termini of the antisense strand, or one or both termini of both strands, wherein the terminal region is the last 5 nucleotides of a strand.

A dsRNA comprising,

• (a) a sense strand having:
• (i) a length of 21 nucleotides;
• (ii) an asialoglycoprotein receptor (ASGPR) ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;
• (iii) 2′-O-Methyl (2′-OMe) modifications at positions 1 to 6, 8, and 12 to 21, and 2′-deoxy-2′-fluoro (2′-F) modifications at positions 7, and 9 to 11;
• counting from the 5′-end of the sense strand;
• and
• (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3, counting from the 5′-end of the sense strand;
• 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 of the antisense strand;
• 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 of the antisense strand;
• wherein the dsRNA has a two-nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.