Claims for Patent: 10,590,418

Summary for Patent: 10,590,418

Title:	Methods and compositions for RNAi mediated inhibition of gene expression in mammals
Abstract:	Methods and compositions are provided for modulating, e.g., reducing, viral coding sequence expression in mammals and mammalian cells. In the subject methods, an effective amount of an RNAi agent, e.g., an interfering ribonucleic acid (such as an siRNA or shRNA) or a transcription template thereof, e.g., a DNA encoding an shRNA, is introduced into a target cell, e.g., by being administered to a mammal that includes the target cell, e.g., via a hydrodynamic administration protocol. Also provided are RNAi agent pharmaceutical preparations for use in the subject methods. The subject methods and compositions find use in a variety of different applications, including academic and therapeutic applications.
Inventor(s):	Kay; Mark A. (Los Altos, CA), McCaffrey; Anton (Pacifica, CA)
Assignee:	The Board of Trustees of the Leland Stanford Junior University (Stanford, CA)
Application Number:	10/259,226
Patent Claims:	1. A method of reducing expression of a target RNA sequence in a liver of a non-embryonic mammal, said method comprising: administering to a non-embryonic mammal, an effective amount of an siRNA to reduce expression of a target RNA sequence in liver cells in the mammal in vivo, wherein the siRNA is specific for the target RNA sequence and comprises a duplex structure having a length in a range of from 19 to 29 base pairs, wherein said administering is subcutaneous administration and wherein the siRNA is not administered using a virus or using a template that encodes the siRNA. 2. The method according to claim 1, wherein the non-embryonic mammal is an adult. 3. The method according to claim 1, wherein the target RNA sequence is endogenous to the liver cells. 4. The method according to claim 1, wherein the target RNA sequence comprises a pathogen sequence. 5. A method of reducing expression of a target RNA sequence in a liver of a non-embryonic mammal, said method comprising: administering to a non-embryonic mammal, an effective amount of an siRNA to reduce expression of a target RNA sequence in liver cells in the mammal in vivo, wherein the siRNA is specific for the target RNA sequence and comprises a duplex structure that has a length in a range of from 21 to 29 base pairs, wherein said administering is subcutaneous administration and wherein the siRNA is not administered using a virus or using a template that encodes the siRNA. 6. The method according to claim 5, wherein the non-embryonic mammal is an adult. 7. The method according to claim 5, wherein the target RNA is sequence endogenous to the liver cells. 8. The method according to claim 5, wherein the target RNA sequence comprises a pathogen sequence. 9. A method of reducing expression of a target RNA sequence in a liver of a non-embryonic mammal, said method comprising: administering to a non-embryonic mammal, an effective amount of an siRNA to reduce expression of a target RNA sequence in liver cells in the mammal in vivo, wherein the siRNA is specific for the target RNA sequence and comprises a duplex structure of 21 base pairs in length, wherein said administering is subcutaneous administration and wherein the siRNA is not administered using a virus or using a template that encodes the siRNA. 10. The method according to claim 1, wherein siRNA is chemically modified. 11. The method according to claim 10, wherein the chemically modified siRNA comprises one or more backbone modifications selected from the group consisting of: a phosphorodiamidate, a methylphosphonate, a phosphorothioate, a phosphorodithioate, a phosphoroamidite, an alkyl phosphotriester, and a boranophosphate. 12. The method according to claim 10, wherein the chemically modified siRNA comprises a sugar modification. 13. The method according to claim 10, wherein the chemically modified siRNA comprises a 2'-O-methyl sugar modification. 14. The method according to claim 5, wherein siRNA is chemically modified. 15. The method according to claim 14, wherein the chemically modified siRNA comprises one or more backbone modifications selected from the group consisting of: a phosphorodiamidate, a methylphosphonate, a phosphorothioate, a phosphorodithioate, a phosphoroamidite, an alkyl phosphotriester, and a boranophosphate. 16. The method according to claim 14, wherein the chemically modified siRNA comprises a sugar modification. 17. The method according to claim 14, wherein the chemically modified siRNA comprises a 2'-O-methyl sugar modification. 18. The method according to claim 17, wherein the non-embryonic mammal is an adult. 19. The method according to claim 17, wherein the target RNA sequence is endogenous to the liver cells. 20. The method according to claim 17, wherein siRNA is chemically modified. 21. The method according to claim 20, wherein the chemically modified siRNA comprises one or more backbone modifications selected from the group consisting of: a phosphorodiamidate, a methylphosphonate, a phosphorothioate, a phosphorodithioate, a phosphoroamidite, an alkyl phosphotriester, and a boranophosphate. 22. The method according to claim 20, wherein the chemically modified siRNA comprises a sugar modification. 23. The method according to claim 20, wherein the chemically modified siRNA comprises a 2'-O-methyl sugar modification. 24. A method of reducing expression of a target RNA sequence in a liver of a non-embryonic mammal, said method comprising: administering to a non-embryonic mammal, an effective amount of an siRNA to reduce expression of a target RNA sequence in liver cells in the mammal in vivo, wherein the siRNA is specific for the target RNA sequence and comprises a duplex structure having a length in a range of from 19 to 29 base pairs, wherein said administering is intravenous administration and wherein the siRNA is not administered using a virus or using a template that encodes the siRNA. 25. The method according to claim 24, wherein the non-embryonic mammal is an adult. 26. The method according to claim 24, wherein the target RNA sequence is endogenous to the liver cells. 27. The method according to claim 24, wherein siRNA is chemically modified. 28. The method according to claim 27, wherein the chemically modified siRNA comprises one or more backbone modifications selected from the group consisting of: a phosphorodiamidate, a methylphosphonate, a phosphorothioate, a phosphorodithioate, a phosphoroamidite, an alkyl phosphotriester, and a boranophosphate. 29. The method according to claim 27, wherein the chemically modified siRNA comprises a sugar modification. 30. The method according to claim 27, wherein the chemically modified siRNA comprises a 2'-O-methyl sugar modification. 31. A method of reducing expression of a target RNA sequence in a liver of a non-embryonic mammal, said method comprising: administering to a non-embryonic mammal, an effective amount of an siRNA to reduce expression of a target RNA sequence in liver cells in the mammal in vivo, wherein the siRNA is specific for the target RNA sequence and comprises a duplex structure of 19 base pairs in length, wherein said administering is intravenous administration and wherein the siRNA is not administered using a virus or using a template that encodes the siRNA. 32. The method according to claim 31, wherein the non-embryonic mammal is an adult. 33. The method according to claim 31, wherein the target RNA sequence is endogenous to the liver cells. 34. The method according to claim 31, wherein siRNA is chemically modified. 35. The method according to claim 34, wherein the chemically modified siRNA comprises one or more backbone modifications selected from the group consisting of: a phosphorodiamidate, a methylphosphonate, a phosphorothioate, a phosphorodithioate, a phosphoroamidite, an alkyl phosphotriester, and a boranophosphate. 36. The method according to claim 34, wherein the chemically modified siRNA comprises a sugar modification. 37. The method according to claim 34, wherein the chemically modified siRNA comprises a 2'-O-methyl sugar modification.