RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA)
View Patent ↗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 ribonucleic acid (siRNA) duplex consisting of a sense strand and an antisense strand, wherein:
(a) each strand is 18 nucleotides in length;
(b) the siRNA duplex has 18 base pairs;
(c) the antisense strand is complementary to a target RNA encoded by a mammalian gene;
(d) the sense strand and the antisense strand are 100% complementary; and
(e) each strand of the siRNA duplex comprises 10 or more 2′-O-methyl nucleotides.
2. A double stranded short interfering ribonucleic acid (siRNA) duplex consisting of a sense strand and an antisense strand, wherein:
(a) each strand is 19 nucleotides in length;
(b) the siRNA duplex has 19 base pairs;
(c) the antisense strand is complementary to a target RNA encoded by a mammalian gene;
(d) the sense strand and the antisense strand are 100% complementary; and
(e) each strand of the siRNA duplex comprises 10 or more 2′-O-methyl nucleotides.
3. A double stranded short interfering ribonucleic acid (siRNA) duplex consisting of a sense strand and an antisense strand, wherein:
(a) each strand is 20 nucleotides in length;
(b) the siRNA duplex has 20 base pairs;
(c) the antisense strand is complementary to a target RNA encoded by a mammalian gene;
(d) the sense strand and the antisense strand are 100% complementary; and
(e) each strand of the siRNA duplex comprises 10 or more 2′-O-methyl nucleotides.
4. A double stranded short interfering ribonucleic acid (siRNA) duplex consisting of a sense strand and an antisense strand, wherein:
(a) each strand is 21 nucleotides in length;
(b) the siRNA duplex has 21 base pairs;
(c) the antisense strand is complementary to a target RNA encoded by a mammalian gene;
(d) the sense strand and the antisense strand are 100% complementary; and
(e) each strand of the siRNA duplex comprises 10 or more 2′-O-methyl nucleotides.
5. A double stranded short interfering ribonucleic acid (siRNA) duplex consisting of a sense strand and an antisense strand, wherein:
(a) each strand is 22 nucleotides in length;
(b) the siRNA duplex has 22 base pairs;
(c) the antisense strand is complementary to a target RNA encoded by a mammalian gene;
(d) the sense strand and the antisense strand are 100% complementary; and
(e) each strand of the siRNA duplex comprises 10 or more 2′-O-methyl nucleotides.
6. A double stranded short interfering ribonucleic acid (siRNA) duplex consisting of a sense strand and an antisense strand, wherein:
(a) each strand is 23 nucleotides in length;
(b) the siRNA duplex has 23 base pairs;
(c) the antisense strand is complementary to a target RNA encoded by a mammalian gene;
(d) the sense strand and the antisense strand are 100% complementary; and
(e) each strand of the siRNA duplex comprises 10 or more 2′-O-methyl nucleotides.
7. The siRNA duplex of claim 1 , wherein each strand of the siRNA duplex further comprises 5 or more 2′-deoxy-2′-fluoro nucleotides.
8. The siRNA duplex of claim 2 , wherein each strand of the siRNA duplex further comprises 5 or more 2′-deoxy-2′-fluoro nucleotides.
9. The siRNA duplex of claim 3 , wherein each strand of the siRNA duplex further comprises 5 or more 2′-deoxy-2′-fluoro nucleotides.
10. The siRNA duplex of claim 4 , wherein each strand of the siRNA duplex further comprises 5 or more 2′-deoxy-2′-fluoro nucleotides.
11. The siRNA duplex of claim 5 , wherein each strand of the siRNA duplex further comprises 5 or more 2′-deoxy-2′-fluoro nucleotides.
12. The siRNA duplex of claim 6 , wherein each strand of the siRNA duplex further comprises 5 or more 2′-deoxy-2′-fluoro nucleotides.
13. The siRNA duplex of claim 1 , wherein each strand of the siRNA duplex further comprises 5 or more phosphorothioate internucleotide linkages.
14. The siRNA duplex of claim 2 , wherein each strand of the siRNA duplex further comprises 5 or more phosphorothioate internucleotide linkages.
15. The siRNA duplex of claim 3 , wherein each strand of the siRNA duplex further comprises 5 or more phosphorothioate internucleotide linkages.
16. The siRNA duplex of claim 4 , wherein each strand of the siRNA duplex further comprises 5 or more phosphorothioate internucleotide linkages.
17. The siRNA duplex of claim 5 , wherein each strand of the siRNA duplex further comprises 5 or more phosphorothioate internucleotide linkages.
18. The siRNA duplex of claim 6 , wherein each strand of the siRNA duplex further comprises 5 or more phosphorothioate internucleotide linkages.
19. A composition comprising the siRNA duplex of claim 1 and a pharmaceutically acceptable carrier or diluent.
20. A composition comprising the siRNA duplex of claim 2 and a pharmaceutically acceptable carrier or diluent.
21. A composition comprising the siRNA duplex of claim 3 and a pharmaceutically acceptable carrier or diluent.
22. A composition comprising the siRNA duplex of claim 4 and a pharmaceutically acceptable carrier or diluent.
23. A composition comprising the siRNA duplex of claim 5 and a pharmaceutically acceptable carrier or diluent.
24. A composition comprising the siRNA duplex of claim 6 and a pharmaceutically acceptable carrier or diluent.