Claims
- 1. A recombinant viral vector comprising:
a chimeric capsid having at least one non-native amino acid sequence, wherein the non-native amino acid sequence is derived from a capsid protein domain of a parvovirus, a virus, or a combination thereof, and wherein the chimeric capsid is capable of binding to an attachment site present on a cell surface; and a transgene flanked 5′ and 3′ by inverted terminal repeat sequences, wherein the inverted terminal repeat sequences are derived from a parvovirus, a virus, or a combination thereof, and wherein at least one inverted terminal repeat sequence comprises a packaging signal that allows assembly of the chimeric capsid.
- 2. The recombinant viral vector of claim 1, wherein the chimeric capsid has a modified tropism.
- 3. The recombinant viral vector of claim 2, wherein the chimeric capsid with a modified tropism permits binding of the viral vector to an attachment site on a cell surface with higher affinity than a corresponding viral vector with a wild type capsid.
- 4. The recombinant viral vector of claim 1, wherein the parvovirus selected from the group consisting of AAV-1, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6.
- 5. The recombinant viral vector of claim 4, wherein the parvovirus comprises a capsid protein with viral protein domain selected from the group consisting of VP1, VP2 and VP3.
- 6. The recombinant viral vector of claim 1, wherein the non-native amino acid sequence is a combination of amino acid sequences derived from one or more parvoviruses selected from the group consisting of AAV-1, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6.
- 7. The recombinant viral vector of claim 6, wherein the non-native amino acid sequence is a combination of an amino acid sequence derived from AAV-2 and an amino acid sequence derived from AAV-5.
- 8. The recombinant viral vector of claim 1, wherein the non-native amino acid sequence is derived from a virus.
- 9. The recombinant viral vector of claim 8, wherein the virus is selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 10. The recombinant viral vector of claim 1, wherein the non-native amino acid sequence is a combination of at least one amino acid sequence derived from a parvovirus selected from the group consisting of AAV-1, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6, and at least one amino acid sequence derived from a virus selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 11. The recombinant viral vector of claim 1, wherein the inverted terminal repeat sequences are each derived from a parvovirus selected from the group consisting of AAV-1, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6.
- 12. The recombinant viral vector of claim 1, wherein the inverted terminal repeat sequences are each derived from a viruses selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 13. The recombinant viral vector of claim 1, wherein the inverted terminal repeat sequences are a combination of at least one inverted terminal repeat sequence derived from a parvovirus selected from the group consisting of AAV-1, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6, and at least one inverted terminal repeat sequence derived from a virus selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 14. The recombinant viral vector of claim 1, wherein the transgene is selected from the group consisting of an RNA molecule, a DNA molecule, and a synthetic DNA molecule.
- 15. A recombinant AAV-2 vector comprising:
a chimeric capsid having at least one native AAV-2 amino acid sequence, and at least one non-native amino acid sequence derived from a parvovirus other than AAV-2, wherein the chimeric capsid is capable of binding to an attachment site present on a cell surface; and a transgene flanked 5′ and 3′ by a first inverted terminal repeat sequence derived from AAV-2 and a second inverted terminal repeat sequence derived from a parvovirus.
- 16. The recombinant AAV-2 vector of claim 15, wherein the chimeric capsid has a modified tropism.
- 17. The recombinant AAV-2 vector of claim 16, wherein the chimeric capsid with a modified tropism permits binding of the AAV-2 vector to an attachment site on a cell surface with higher affinity than that exhibited by a corresponding AAV-2 vector with a wild type AAV-2 capsid.
- 18. The recombinant AAV-2 vector of claim 15, wherein the amino acid sequence derived from AAV-2 comprises a viral protein domain selected from the group consisting of VP1, VP2 and VP3.
- 19. The recombinant AAV-2 vector of claim 15, wherein the non-native amino acid sequence is derived from a parvovirus selected from the group consisting of AAV-1, AAV-3, AAV-5 and AAV-6.
- 20. The recombinant AAV-2 vector of claim 19, wherein the non-native amino acid sequence of the parvovirus comprises a viral protein domain selected from the group consisting of VP1, VP2 and VP3.
- 21. The recombinant AAV-2 vector of claim 15, wherein the chimeric capsid comprises a native amino acid sequence derived from the VP1 domain of AAV-2 and, wherein the non-native amino acid sequence comprises a VP2 domain and a VP3 domain derived from AAV-5.
- 22. The recombinant AAV-2 vector of claim 15, wherein the second inverted terminal repeat sequence derived from a parvovirus is selected from the group consisting of AAV-1, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6.
- 23. The recombinant AAV-2 vector of claim 15, wherein the transgene is selected from the group consisting of an RNA molecule, a DNA molecule, and a synthetic DNA molecule.
- 24. A recombinant AAV-2 vector comprising:
a chimeric capsid having at least one native AAV-2 amino acid sequence and at least one non-native amino acid sequence derived from a virus, wherein the chimeric capsid is capable of binding to an attachment site present on a cell surface; and a transgene flanked 5′ and 3′ by a first inverted terminal repeat sequence derived from AAV-2 and a second inverted terminal repeat sequence derived from a parvovirus.
- 25. The recombinant AAV-2 vector of claim 24, wherein the chimeric capsid has a modified tropism.
- 26. The recombinant AAV-2 vector of claim 25, wherein the chimeric capsid with a modified tropism permits binding of the AAV-2 vector to an attachment site on a cell surface with higher affinity than a corresponding AAV-2 vector with a wild type capsid.
- 27. The recombinant AAV-2 vector of claim 24, wherein the amino acid sequence derived from AAV-2 comprises a viral protein domain selected from the group consisting of VP1, VP2 and VP3.
- 28. The recombinant AAV-2 vector of claim 24, wherein the non-native amino acid sequence is derived from a virus selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 29. The recombinant AAV-2 vector of claim 24, wherein the second inverted terminal repeat sequence is derived from a parvovirus selected from the group consisting of AAV-1, AAV-3, AAV-4, AAV-5 and AAV-6.
- 30. The recombinant AAV-2 vector of claim 24, wherein the transgene is selected from the group consisting of an RNA molecule, a DNA molecule, and a synthetic DNA molecule.
- 31. A recombinant AAV-2 vector comprising:
a chimeric capsid having at least one native AAV-2 amino acid sequence, and at least one non-native amino acid sequence derived from a virus, wherein the chimeric capsid is capable of binding to an attachment site present on a cell surface; and a transgene flanked by a first inverted terminal repeat sequence derived from AAV-2 and a second inverted terminal repeat sequence derived from a virus.
- 32. The recombinant AAV-2 vector of claim 31, wherein the chimeric capsid has a modified tropism.
- 33. The recombinant AAV-2 vector of claim 32, wherein the chimeric capsid with a modified tropism permits binding of the AAV-2 vector to an attachment site on a cell surface with higher affinity than a corresponding AAV-2 vector with a wild type capsid.
- 34. The recombinant AAV-2 vector of claim 31, wherein the amino acid sequence derived from AAV-2 comprises a viral protein domain selected from the group consisting of VP1, VP2 and VP3.
- 35. The recombinant AAV-2 vector of claim 31, wherein the non-native amino acid sequence is derived from a virus selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 36. The recombinant AAV-2 vector of claim 31, wherein the second terminal repeat sequence is derived from a virus selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 37. The recombinant AAV-2 vector of claim 31, wherein the transgene is selected from the group consisting of an RNA molecule, a DNA molecule, and a synthetic DNA molecule.
- 38. A chimeric capsid vehicle comprising a native AAV-2 amino acid sequence and at least one non-native amino acid sequence derived from a capsid protein of a parvovirus other than AAV-2, covalently linked to a transgene.
- 39. The chimeric capsid vehicle of claim 38 wherein the chimeric capsid has a modified tropism.
- 40. The chimeric capsid vehicle of claim 39, wherein the chimeric capsid with a modified tropism permits binding of the chimeric capsid to an attachment site on a cell surface with higher affinity than a corresponding wild type capsid vehicle.
- 41. The chimeric capsid vehicle of claim 38, wherein the amino acid sequence derived from AAV-2 comprises a viral protein domain selected from the group consisting of VP 1, VP2 and VP3.
- 42. The chimeric capsid vehicle of claim 38, wherein the non-native amino acid sequence is derived from a parvovirus selected from the group consisting of AAV-1, AAV-3, AAV-5 and AAV-6.
- 43. The chimeric capsid vehicle of claim 38, wherein the transgene is selected from the group consisting of an RNA molecule, a DNA molecule, and a synthetic DNA molecule.
- 44. A chimeric capsid vehicle comprising a native AAV-2 amino acid sequence and at least one non-native amino acid derived from a capsid protein of a virus, covalently linked to a transgene.
- 45. The chimeric capsid vehicle of claim 44 wherein the chimeric capsid has a modified tropism.
- 46. The chimeric capsid vehicle of claim 45, wherein the chimeric capsid with a modified tropism permits binding of the chimeric capsid to an attachment site on a cell surface with higher affinity than a corresponding wild type capsid vehicle.
- 47. The chimeric capsid vehicle of claim 44, wherein the amino acid sequence derived from AAV-2 comprises a viral protein domain selected from the group consisting of VP1, VP2 and VP3.
- 48. The chimeric capsid vehicle of claim 44, wherein the non-native amino acid sequence is derived from a virus selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 49. The chimeric capsid vehicle of claim 44, wherein the transgene is selected from the group consisting of an RNA molecule, a DNA molecule, and a synthetic DNA molecule.
- 50. A method for modifying the tropism of a recombinant AAV-2 vector comprising:
replacing at least a portion of a native amino acid sequence of an AAV-2 capsid protein with a non-native amino acid sequence derived from a capsid protein of a parvovirus other than AAV-2; and combining the capsid proteins under conditions for assembly to produce a chimeric capsid encapsidating an AAV-2 vector, to thereby modify the tropism of an AAV-2 vector.
- 51. The method of claim 50, wherein the parvovirus is selected from the group consisting of AAV-1, AAV-3, AAV-5 and AAV-6.
- 52. A method for modifying the tropism of a recombinant AAV-2 vector comprising:
replacing at least a portion of a native amino acid sequence of an AAV-2 capsid protein with a non-native amino acid sequence derived from a capsid protein of a virus; and combining the capsid protein under conditions for assembly, to thereby modify the tropism of an AAV-2 vector.
- 53. The method of claim 52, wherein the non-native amino acid sequence is derived from a virus selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 54. A method for improving gene therapy in a subject with a disorder comprising:
administering a therapeutically effective amount of a recombinant vector comprising a transgene and a chimeric capsid capable of binding to an attachment site present on a cell surface; targeting a cell that recombinant vector with a chimeric capsid can bind to with a higher affinity than the corresponding viral vector with a wild type capsid; and expressing the transgene in a subject at a level sufficient to ameliorate the disorder, thereby improving gene therapy.
- 55. The method of claim 54, wherein the step of administering the recombinant vector with a chimeric capsid further comprises administering a recombinant vector comprising a chimeric capsid with at least one amino acid sequence derived from a first parvovirus and at least one amino acid sequence derived from a second parvovirus.
- 56. The method of claim 55, wherein the first parvovirus is selected from the group consisting of AAV-l, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6.
- 57. The method of claim 55, wherein the second parvovirus is selected from the group consisting of AAV-l, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6.
- 58. The method of claim 54, wherein the step of administering the recombinant vector with a chimeric capsid comprises administering a recombinant vector comprising a chimeric capsid with at least one amino acid sequence derived from a parvovirus and at least one amino acid sequence derived from a virus.
- 59. The method of claim 58, wherein the parvovirus is selected from the group consisting of AAV-1, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6.
- 60. The method of claim 58, wherein the virus is selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 61. The method of claim 54, wherein the step of administering the recombinant vector with a chimeric capsid comprises administering a recombinant vector comprising a chimeric capsid with at least one amino acid sequence derived from AAV-2 and at least one amino acid sequence derived from a parvovirus.
- 62. The method of claim 61, wherein the parvovirus is selected from the group consisting of AAV-1, AAV-3, AAV-5 and AAV-6.
- 63. The method of claim 54, wherein the step of administering the recombinant vector with a chimeric capsid comprises administering a recombinant vector comprising a chimeric capsid with at least one amino acid sequence derived from AAV-2 and at least one amino acid sequence derived from a virus.
- 64. The method of claim 63, wherein the virus is selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 65. A method for increasing the efficiency of entry into a cell using a recombinant viral vector with a chimeric capsid comprising:
producing a chimeric capsid encapsidating a viral vector, wherein the chimeric capsid has a modified tropism; and contacting a cell with the recombinant viral vector having a chimeric capsid such that the chimeric capsid binds to an attachment site on the cell surface and permits the vector to enter the cell more efficiently that a viral vector comprising a wild type capsid.
- 66. The method of claim 65, wherein the step of producing a chimeric capsid encapsidating a viral vector comprises producing a chimeric capsid with at least one amino acid sequence derived from a first parvovirus and at least one amino acid sequence derived from a second parvovirus.
- 67. The method of claim 66, wherein the first parvovirus is selected from the group consisting of AAV-1, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6.
- 68. The method of claim 66, wherein the second parvovirus is selected from the group consisting of AAV-1, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6.
- 69. The method of claim 65, wherein the step of producing a chimeric capsid encapsidating a viral vector comprises producing a chimeric capsid with at least one amino acid sequence derived from a parvovirus and at least one amino acid sequence derived from virus.
- 70. The method of claim 69, wherein the parvovirus is selected from the group consisting of AAV-1, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6.
- 71. The method of claim 69, wherein the virus is selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 72. The method of claim 65, wherein the step of producing a chimeric capsid encapsidating a viral vector comprises producing a chimeric capsid with at least one amino acid sequence derived from AAV-2 and at least one amino acid sequence derived from a parvovirus.
- 73. The method of claim 72, wherein the parvovirus is selected from the group consisting of AAV-1, AAV-3, AAV-5 and AAV-6.
- 74. The method of claim 65, wherein the step of producing a chimeric capsid encapsidating a viral vector comprises producing a chimeric capsid with at least one amino acid sequence derived from AAV-2 and at least one amino acid sequence derived from a virus.
- 75. The method of claim 74, wherein the virus is selected from the group consisting of herpesvirus, adenovirus, lentivirus, retrovirus, Epstein-Barr virus and vaccinia virus.
- 76. A method of making a recombinant particle with a chimeric capsid comprising:
providing a first construct comprising a transgene flanked 5′ and 3′ with inverted terminal repeat sequences, wherein at least one invented terminal repeat sequence comprises a packaging signal, and a second construct comprising a nucleic acid sequence encoding a chimeric capsid; and contacting a population of cells with the first and second constructs, such that the population of cells allows assembly of a recombinant particle, to thereby produce a recombinant particle with a chimeric capsid.
- 77. The method of claim 76, wherein the first construct comprises inverted terminal repeat sequences derived from one or more parvoviruses selected from the group consisting of AAV-1, AAV-2, AAV-3, AAV-4, AAV-5 and AAV-6.
- 78. The method of claim 76, wherein the first construct comprises inverted terminal repeat sequences derived from AAV-2.
- 79. The method of claim 76, wherein the second construct further comprises a nucleic acid sequence encoding a chimeric capsid of any one of claims 1, 15, 24 or 31.
- 80. The method of claim 76, wherein the step of contacting the population of cells further comprises contacting a population of 293 cells.
- 81. A cell comprising a recombinant viral vector comprising a chimeric capsid of any of claims 1, 15, 24 or 31.
- 82. A pharmaceutical composition comprising a recombinant viral vector comprising
a chimeric capsid of any one of claims 1, 15, 24 or 31; and a pharmaceutically acceptable carrier.
Parent Case Info
[0001] This application claims priority to U.S. Provisional Patent Application No. 60/189,110, filed Mar. 14, 2000.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60189110 |
Mar 2000 |
US |