Modified small interfering RNA molecules and methods of use
View Patent ↗The present invention provides double-stranded RNA molecules that mediate RNA interference in target cells, preferably hepatic cells. The invention also provides double-stranded RNA molecules that are modified to be resistant to nuclease degradation, which inactivates a virus, and more specifically, hepatitis C virus (HCV). The invention also provides a method of using these modified RNA molecules to inactivate virus in mammalian cells and a method of making modified small interfering RNAs (siRNAs) using human Dicer.
1. A composition comprising a siRNA to HCV, comprising a first strand and a second strand, wherein the sequence of said first strand is the sequence of SEQ ID NO: 46.
2. The composition of claim 1 , wherein the siRNA comprises at least one modification at the 2′ position of at least one ribonucleotide.
3. The composition of claim 1 , wherein said siRNA comprises at least one modification selected from the group consisting of fluoro-, methyl-, methoxyethyl- and propyl-modification.
4. The composition of claim 3 , wherein said fluoro-modification is a 2′-fluoro-modification or a 2′,2′-difluoro-modification.
5. The composition of claim 1 , wherein at least one pyrimidine of said siRNA is modified.
6. The composition of claim 1 , wherein both strands of said siRNA contain at least one modified nucleotide.
7. A method for inactivating Hepatitis C Virus (HCV) in a patient, the method comprising the step of administering to said patient a siRNA in an amount effective to inactivate said virus, wherein said siRNA comprises a first strand and a second strand,
wherein the sequence of said first strand is the sequence of SEQ ID NO: 46 and/or the sequence of said second strand is the sequence of SEQ ID NO: 18 or 45.
8. The method of claim 7 , wherein the siRNA comprises at least one modification at the 2′ position of at least one ribonucleotide.
9. The method of claim 7 , wherein said siRNA comprises at least one modification selected from the group consisting of fluoro-, methyl-, methoxyethyl- and propyl-modification.
10. The method of claim 9 , wherein said fluoro-modification is a 2′ fluoro-modification or a 2′,2′-fluoro-modification.
11. The method of claim 7 , wherein at least one pyrimidine of said siRNA is modified.
12. The method of claim 7 , wherein both strands of said siRNA contain at least one modified nucleotide.
13. A vector encoding the first and/or the second strand of the double-stranded RNA of claim 1 .
14. A host cell comprising the vector of claim 13 .
15. The double-stranded RNA molecule of claim 1 , further comprising a receptor-binding ligand attached to a 5′-end or 3′-end of said siRNA molecule.
16. The double-stranded RNA molecule of claim 15 , wherein said receptor-binding ligand is selected from the group consisting of a cholesterol, an HBV surface antigen, low-density lipoprotein, an HIV-1 surface antigen, an influenza virus surface antigen, an RSV surface antigen, an HPV surface antigen and a polio virus surface antigen.
17. The double-stranded RNA molecule of claim 1 , wherein the double-stranded RNA molecule is in a combination with a second siRNA comprising a first strand and a second strand.
18. The composition of claim 1 , wherein the sequence of the second strand is the sequence of SEQ ID NO: 18 or 45.