Methods and compositions for the specific inhibition of alpha-1 antitrypsin by double-stranded RNA
This invention relates to compounds, compositions, and methods useful for reducing α-1 antitrypsin target RNA and protein levels via use of dsRNAs, e.g., Dicer substrate siRNA (DsiRNA) agents.
1. A nucleic acid comprising an oligonucleotide strand consisting of 22 nucleotides in length, wherein said oligonucleotide strand has a region that is perfectly complementary to a contiguous sequence of α-1 antitrypsin mRNA consisting of 20 nucleotides in length, which contiguous sequence comprises a target α-1 antitrypsin mRNA sequence as set forth in SEQ ID NO: 1547, and wherein the nucleic acid comprises one or more modified nucleotides.
2. A mammalian cell containing the nucleic acid of claim 1 .
3. A pharmaceutical composition comprising the nucleic acid of claim 1 and a pharmaceutically acceptable carrier.
4. A kit comprising the nucleic acid of claim 1 and instructions for its use.
5. The nucleic acid of claim 1 , further comprising one or more phosphorothioate linkages.
6. A double stranded nucleic acid (dsNA) comprising a sense strand and an antisense strand, wherein the antisense strand is 22 nucleotides in length, wherein the antisense strand has a region that is perfectly complementary to a contiguous sequence of α-1 antitrypsin mRNA consisting of 20 nucleotides in length, which contiguous sequence comprises a target α-1 antitrypsin mRNA sequence as set forth in SEQ ID NO: 1547, and wherein the dsNA comprises one or more modified nucleotides.
7. The dsNA of claim 6 , wherein the sense strand and antisense strand form a duplex such that the antisense strand comprises a two nucleotide overhang at its 3′ end.
8. The dsNA of claim 6 , wherein the dsNA comprises a duplex of 20 to 25 base pairs in length.
9. The dsNA of claim 7 , wherein the sense strand is 25 to 53 nucleotides in length.
10. The dsNA of claim 7 , wherein the sense strand is 36 nucleotides in length.
11. The dsNA of claim 7 , wherein the 3′ terminus of the sense strand and the 5′ terminus of the antisense strand form a blunt end.
12. The dsNA of claim 6 , wherein the sense strand comprises a tetraloop at its 3′ end.
13. The dsNA of claim 10 , wherein the sense strand comprises a tetraloop at its 3′ end.
14. The dsNA of claim 13 , wherein the tetraloop comprises a single stranded loop having a sequence of GAAA.
15. The dsNA of claim 6 , wherein the dsNA comprises at least one phosphorothioate linkage.
16. The dsNA of claim 14 , wherein the dsNA comprises at least one phosphorothioate linkage.
17. The dsNA of claim 6 , wherein the one or more modified nucleotides contains a 2′ modification.
18. The dsNA of claim 17 , wherein the one or more modified nucleotides are selected from a 2′-O-methyl nucleotide and a 2′-fluoro nucleotide.
19. The dsNA of claim 18 , wherein the dsNA comprises at least one 2′-fluoro nucleotide and at least one 2′-O-methyl nucleotide.
20. The dsNA of claim 6 , wherein the dsNA is attached to a moiety selected from the group consisting of a GalNAc moiety, a cholesterol moiety, and a cholesterol targeting ligand.
21. The dsNA of claim 18 , wherein the dsNA is attached to a GalNAc moiety.
22. A pharmaceutical composition comprising the dsNA of claim 21 and a pharmaceutically acceptable carrier.