RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA)
The present invention concerns methods and reagents useful in modulating gene expression in a variety of applications, including use in therapeutic, diagnostic, target validation, and genomic discovery applications. Specifically, the invention relates to synthetic chemically modified small nucleic acid molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA) molecules capable of mediating RNA interference (RNAi) against target nucleic acid sequences. The small nucleic acid molecules are useful in the treatment of any disease or condition that responds to modulation of gene expression or activity in a cell, tissue, or organism.
1. A short interfering RNA (siRNA) molecule having a sense strand and an antisense strand that mediates RNA interference against a Hepatitis B Virus target RNA, wherein:
(a) each strand is between 18 and 24 nucleotides in length;
(b) the sense strand comprises 10 or more 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, or universal base modified nucleotides;
(c) the antisense strand comprises 10 or more 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, or universal base modified nucleotides; and
(d) 10 or more pyrimidine nucleotides of the sense and/or antisense strand are 2′-deoxy, 2′-O-methyl or 2′-deoxy-2′-fluoro nucleotides.
2. The siRNA molecule of claim 1 , wherein the sense strand comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 phosphorothioate internucleotide linkages.
3. The siRNA molecule of claim 1 , wherein the antisense strand comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 phosphorothioate internucleotide linkages.
4. A composition comprising the siRNA molecule of claim 1 and a pharmaceutically acceptable carrier or diluent.