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 double-stranded short interfering nucleic acid (siNA) molecule comprising a sense strand and an antisense strand, wherein:
a) the antisense strand is complementary to the nucleotide sequence of SEQ ID NO: 449;
b) each strand of said double-stranded siNA molecule is 21 nucleotides in length;
c) all 21 nucleotides of each strand of the siNA molecule are base-paired to the complementary nucleotides of the other strand of the siNA molecule;
(d) the sense strand comprises 10 or more 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, or universal base modified nucleotides;
(e) the antisense strand comprises 10 or more 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, or universal base modified nucleotides; and
(f) 10 or more pyrimidine nucleotides of the sense and/or antisense strand are 2′-deoxy, 2′-O-methyl or 2′-deoxy-2′-fluoro nucleotides; and
wherein the siNA molecule downregulates expression of a Hepatitis B virus target gene.
2. The siNA molecule of claim 1 , wherein said antisense strand comprises a phosphorothioate internucleotide linkage at the 3′ end of said antisense strand.
3. The siNA molecule of claim 1 , which comprises no ribonucleotides.
4. A pharmaceutical composition comprising the siNA molecule of claim 1 in an acceptable carrier or diluent.