Claims
- 1. A cDNA molecule encoding a fusion protein that comprises mammalian DHFR and a therapeutic protein.
- 2. The cDNA of claim 1, wherein the fusion protein comprises a wild-type mammalian DHFR.
- 3. The cDNA of claim 2, wherein the wild-type mammalian DHFR is rat, mouse, dog, monkey or human DHFR.
- 4. The cDNA of claim 1, wherein the fusion protein comprises a mutant form of DHFR.
- 5. The cDNA of claim 4, wherein the fusion protein comprises a mutant form of human DHFR.
- 6. The cDNA of claim 5, wherein the mutant form of human DHFR has increased resistance to methotrexate.
- 7. The cDNA of claim 4, wherein the mutant form of human DHFR differs from wild-type DHFR as a result of one or more mutations, including at least one mutation at an amino acid corresponding to amino acid 15, 22, 31 or 34 of the wild-type sequence.
- 8. The eDNA of claim 7, wherein the mutant form of human DHFR differs from wild-type human DHFR as a result of a set of mutations comprising a mutation at the amino acid corresponding to amino acid 22 and a mutation at the amino acid corresponding to amino acid 31 of the wild-type sequence.
- 9. The cDNA of claim 1, wherein the therapeutic protein is a protein that enhances toxicity of an administered drug.
- 10. The cDNA of claim 9, wherein the therapeutic protein is a mutant or wild-type form of herpes simplex virus thymidine kinase.
- 11. The cDNA of claim 10, wherein the fusion protein comprises a wild-type mammalian DHFR.
- 12. The cDNA of claim 11, wherein the wild-type mammalian DHFR is rat, mouse, dog, monkey or human DHFR.
- 13. The cONA of claim 10, wherein the fusion protein comprises a mutant form of DHFR.
- 14. The cDNA of claim 13, wherein the fusion protein comprises a mutant form of human DHFR.
- 15. The cDNA of claim 14, wherein the mutant form of human DHFR has increased resistance to methotrexate.
- 16. The cDNA of claim 13, wherein the mutant form of human DHFR differs from wild-type DHFR as a result of one or more mutations, including at least one mutation at an amino acid corresponding to amino acid 15, 22, 31 or 34 of the wild-type sequence.
- 17. The cDNA of claim 16, wherein the mutant form of human DHFR differs from wild-type human DHFR as a result of a set of mutations comprising a mutation at the amino acid corresponding to amino acid 22 and a mutation at the amino acid corresponding to amino acid 31 of the wild-type sequence.
- 18. The cDNA of claim 9, wherein the therapeutic protein is cytosine deaminase.
- 19. The cDNA of claim 1, wherein the therapeutic protein is a product of a pro-apoptotic gene.
- 20. The cDNA of claim 19, wherein the fusion protein comprises a wild-type mammalian DHFR.
- 21. The cDNA of claim 19, wherein the fusion protein comprises a mutant form of DHFR.
- 22. The cDNA of claim 21, wherein the fusion protein comprises a mutant form of human DHFR.
- 23. The cDNA of claim 22, wherein the mutant form of human DHFR has increased resistance to methotrexate.
- 24. The cDNA of claim 22, wherein the mutant form of human DHFR differs from wild-type DHIFR as a result of one or more mutations, including at least one mutation at an amino acid corresponding to amino acid 15, 22, 31 or 34 of the wild-type sequence.
- 25. The cDNA of claim 24, wherein the mutant form of human DHFR differs from wild-type human DHFR as a result of a set of mutations comprising a mutation at the amino acid corresponding to amino acid 22 and a mutation at the amino acid corresponding to amino acid 31 of the wild-type sequence.
- 26. The cDNA of claim 1, wherein the therapeutic protein is a product of a tumor suppressor gene.
- 27. The cDNA of claim 19, wherein the fusion protein comprises a wild-type mammalian DHFR.
- 28. The cDNA of claim 27, wherein the fusion protein comprises a mutant form of DHFR.
- 29. The cDNA of claim 28, wherein the fusion protein comprises a mutant fonrm of human DHFR.
- 30. The cDNA of claim 29, wherein the mutant form of human DHFR has increased resistance to methotrexate.
- 31. The cDNA of claim 29, wherein the mutant form of human DHFR differs from wild-type DHFR as a result of one or more mutations, including at least one mutation at an amino acid corresponding to amino acid 15, 22, 31 or 34 of the wild-type sequence.
- 32. The cDNA of claim 31, wherein the mutant form of human DHFR differs from wild-type human DHFR as a result of a set of mutations comprising a mutation at the amino acid corresponding to amino acid 22 and a mutation at the amino acid corresponding to amino acid 31 of the wild-type sequence.
- 33. The cDNA of claim 1, wherein the therapeutic protein is an immunostimulatory molecule.
- 34. The cDNA of claim 33, wherein the fusion protein comprises a wild-type mammalian DHFR.
- 35. The cDNA of claim 33, wherein the fusion protein comprises a mutant form of DHFR.
- 36. The cDNA of claim 35, wherein the fusion protein comprises a mutant form of human DHFR.
- 37. The cDNA of claim 36, wherein the mutant form of human DHFR has increased resistance to methotrexate.
- 38. The cDNA of claim 36, wherein the mutant form of human DHFR differs from wild-type DHFR as a result of one or more mutations, including at least one mutation at anamino acid corresponding to amino acid 15, 22, 31 or 34 of the wild-type sequence.
- 39. The cDNA of claim 38, wherein the mutant form of human DHFR differs from wild-type human DHFR as a result of a set of mutations comprising a mutation at the amino acid corresponding to amino acid 22 and a mutation at the amino acid corresponding to amino acid 31 of the wild-type sequence.
- 40. The cDNA of claim 1, wherein the therapeutic protein is a functional protein useful in gene therapy.
- 41. The cDNA of claim 40, wherein the therapeutic protein is a wild-type or enhanced beta-globin protein.
- 42. The cDNA of claim 1, further comprising a region encoding a reporter protein.
- 43. The cDNA of claim 42, wherein the reporter protein is green fluorescent protein.
- 44. A fusion protein that comprises mammalian DHFR and a therapeutic protein.
- 45. The fusion protein of claim 44, wherein the fusion protein comprises a wild-type mammalian DHFR.
- 46. The fusion protein claim 46, wherein the wild-type mammalian DHFR is rat, mouse, dog, monkey or human DHFR.
- 47. The fusion protein of claim 44, wherein the fusion protein comprises a mutant form of DHFR.
- 48. The fusion protein of claim 47, wherein the fusion protein comprises a mutant form of human DHFR.
- 49. The fusion protein of claim 48, wherein the mutant form of human DHFR has increased resistance to methotrexate.
- 50. The fusion protein of claim 47, wherein the mutant form of human DHFR differs from wild-type DHFR as a result of one or more mutations, including at least one mutation at an amino acid corresponding to amino acid 15, 22, 31 or 34 of the wild-type sequence.
- 51. The fusion protein of claim 50, wherein the mutant form of human DHFR differs from wild-type human DHFR as a result of a set of mutations comprising a mutation at the amino acid corresponding to amino acid 22 and a mutation at the amino acid corresponding to amino acid 31 of the wild-type sequence.
- 52. The fusion protein of claim 44, wherein the therapeutic protein is a protein that enhances toxicity of an administered drug.
- 53. The fusion protein of claim 52, wherein the therapeutic protein is a mutant or wild-type form of herpes simplex virus thymidine kinase.
- 54. The fusion protein of claim 52, wherein the therapeutic protein is cytosine deaminase.
- 55. The fusion protein of claim 44, wherein the therapeutic protein is a product of a pro-apoptotic gene.
- 56. The fusion protein of claim 44, wherein the therapeutic protein is a product of a tumor suppressor gene.
- 57. The fusion protein of claim 44, wherein the therapeutic protein is an immunostimulatory molecule.
- 58. The fusion protein of claim 44, wherein the therapeutic protein is a functional protein useful in gene therapy.
- 59. The fusion protein of claim 58, wherein the therapeutic protein is a wild-type or enhanced beta-globin protein.
- 60. The fusion protein of claim 44, further comprising a reporter protein.
- 61. The fusion protein of claim 60, wherein the reporter protein is green fluorescent protein.
- 62. A method for providing enhanced delivery of a therapeutic protein to a mammalian subject comprising the step of administering to the mammalian subject a cDNA molecule encoding a fusion protein that comprises mammalian DHFR and a therapeutic protein such that the cDNA molecule is expressed
- 63. The method of claim 62, wherein the fusion protein comprises a wild-type mammalian DHFR.
- 64. The method of claim 63, wherein the wild-type mammalian DHFR is rat, mouse, dog, monkey or human DHFR.
- 65. The method of claim 62, wherein the fusion protein comprises a mutant form of DHFR.
- 66. The method of claim 65, wherein the fusion protein comprises a mutant form of human DHFR.
- 67. The method of claim 66, wherein the mutant form of human DHFR has increased resistance to methotrexate.
- 68. The method of claim 66, wherein the mutant form of human DHFR differs from wild-type DHFR as a result of one or more mutations, including at least one mutation at an amino acid corresponding to amino acid 15, 22, 31 or 34 of the wild-type sequence.
- 69. The method of claim 68, wherein the mutant form of human DHFR differs from wild-type human DHFR as a result of a set of mutations comprising a mutation at the amino acid corresponding to amino acid 22 and a mutation at the amino acid corresponding to amino acid 31 of the wild-type sequence.
- 70. The method of claim 62, wherein the therapeutic protein is a protein that enhances toxicity of an administered drug.
- 71. The method of claim 70, wherein the therapeutic protein is a mutant or wild-type form of herpes simplex virus thymidine kinase.
- 72. The method of claim 70, wherein the therapeutic protein is cytosine deaminase.
- 73. The method of claim 62, wherein the therapeutic protein is a product of a pro-apoptotic gene.
- 74. The method of claim 62, wherein the therapeutic protein is a product of a tumor suppressor gene.
- 75. The method of claim 62, wherein the therapeutic protein is an immunostimulatory molecule.
- 76. The method of claim 62, wherein the therapeutic protein is a functional protein useful in gene therapy.
- 77. The method of claim 76, wherein the therapeutic protein is a wild-type or enhanced beta-globin protein.
- 78. The method of claim 62, wherein the fusion protein further comprises a reporter protein.
- 79. The method of claim 78, wherein the reporter protein is green fluorescent protein.
Parent Case Info
[0001] This application claims the benefit of U.S. Provisional Application Serial No. 60/375,250 filed Apr. 22, 2002, which application is incorporated herein by reference in its entirety.
Government Interests
[0002] The invention disclosed in this application was supported by grants CA-08010 and CA-86438-02 from the National Cancer Institute. The United States Government may have certain rights in this invention.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60375250 |
Apr 2002 |
US |