Claims
- 1. A method of integrating a nucleotide sequence of interest into a genome of a vertebrate comprising:
a) providing an isolated early stage embryo having a plasma membrane; and, b) contacting said plasma membrane with a composition comprising at least a first lentivirus vector comprising the nucleotide sequence of interest.
- 2. The method of claim 1, wherein
a) said isolated early stage embryo further comprises a zona pellucida, said zona pellucida and said plasma membrane defining a perivitelline space; and, b) contacting said plasma membrane comprises introducing into said perivitelline space said composition.
- 3. The method of claim 1, wherein said early stage embryo is a fertilized ooctye.
- 4. The method of claim 1, wherein said early stage embryo is selected from the group consisting of a 2-cell stage embryo, a 4-cell stage embryo, an 8-cell stage embryo, and a morula.
- 5. The method of claim 2, wherein said composition comprising the first lentiviral vector is introduced into the perivitelline space via microinjection.
- 6. The method of claim 3, wherein said first nucleotide sequence of interest is integrated into the genome of the fertilized oocyte.
- 7. The method of claim 1, wherein said first nucleotide sequence of interest encodes a polypeptide.
- 8. The method of claim 1, wherein the first lentivirus vector is derived from a virus selected from the group consisting of a human immunodeficiency virus and a simian immunodeficiency virus.
- 9. The method of claim 1, wherein the first lentiviral vector is a trans-viral vector.
- 10. The method of claim 9, wherein the trans-viral vector is a trans-lentiviral vector or a trans-retroviral vector.
- 11. The method of claim 1, wherein the vertebrate is selected from the group consisting of mice, rabbits, sheep, cattle, aves, and rats.
- 12. The method of claim 1, wherein said composition further comprises a second lentivirus vector comprising a second nucleotide sequence of interest.
- 13. The method of claim 12, wherein the first and the second lentivirus vector is derived from a virus selected from the group consisting of a human immunodeficiency virus and a simian immunodeficiency virus.
- 14. The method of claim 12, wherein the first and the second lentiviral vector is a trans-viral vector.
- 15. The method of claim 14, wherein the trans-viral vector is a trans-lentiviral vector or a trans-retroviral vector.
- 16. The method of claim 1, wherein said method further comprises culturing the early stage embryo under conditions that allow formation of a pre-implantation embryo.
- 17. The method of claim 16, further comprising transferring said pre-implantation embryo into a recipient vertebrate and allowing said pre-implantation embryo to develop into at least one vertebrate.
- 18. The method of claim 16, wherein
a) said early stage embryo further comprises a zona pellucida, said zona pellucida and said plasma membrane defining a perivitelline space; and, b) contacting said plasma membrane comprises introducing into said perivitelline space the composition comprising at least said first lentivirus vector comprising the first nucleotide sequence of interest.
- 19. The method of claim 16, wherein said early stage embryo is a fertilized oocyte.
- 20. The method of claim 16, wherein said early stage embryo is a 2-cell embryo, a 4-cell embryo, an 8-cell embryo, or a morula.
- 21. The method of claim 18, further comprising transferring said pre-implantation embryo into a recipient vertebrate and allowing said pre-implantation embryo to develop into at least one genetically modified vertebrate.
- 22. The method of claim 18, wherein said composition comprising the first lentiviral vector is introduced into the perivitelline space via microinjection.
- 23. The method of claim 19, wherein said first nucleotide sequence of interest is integrated into the genome of the fertilized oocyte.
- 24. The method of claim 16, wherein said composition further comprises a second lentivirus vector comprising a second nucleotide sequence of interest.
- 25. The method of claim 24, further comprising transferring said pre-implantation embryo to a recipient vertebrate and allowing said pre-implantation embryo to develop into at least one genetically modified vertebrate.
- 26. A composition comprising an isolated early stage embryo and an effective concentration of at least a first lentivirus vector comprising a nucleotide sequence of interest, wherein said early stage embryo is from a non-human vertebrate.
- 27. The composition of claim 26, wherein said early stage embryo is a fertilized oocyte.
- 28. The compositions of claim 26, wherein said early stage embryo is a 2-cell embryo, a 4-cell embryo, an 8-cell embryo, or a morula.
- 29. The composition of claim 26, wherein
a) said isolated early stage embryo further comprises a zona pellucida, said zona pellucida and said plasma membrane defining a perivitelline space; and, b) said lentivirus vector is in the perivitelline space.
- 30. The composition of claim 29, wherein said first nucleotide sequence of interest encodes a polypeptide.
- 31. The composition of claim 26, wherein the lentivirus vector is derived from a virus selected from the group consisting of a human immunodeficiency virus and a simian immunodeficiency virus.
- 32. The composition of claim 26, wherein the lentiviral vector is a trans-viral vector.
- 33. The composition of claim 32, wherein the trans-viral vector is a trans-lentiviral vector or a trans-retroviral vector.
- 34. The composition of claim 26, wherein the non-human vertebrate is selected from the group consisting of mice, rabbits, sheep, cattle, aves, and rats.
- 35. The composition of claim 26, further comprising an effective concentration of a second lentivirus vector comprising a second nucleotide sequence of interest.
- 36. A method of integrating a nucleotide sequence of interest into the genome of a vertebrate comprising:
a) providing an isolated oocyte having a plasma membrane; and, b) contacting said plasma membrane with a composition comprising at least a first lentivirus vector comprising the nucleotide sequence of interest.
- 37. The method of claim 36, wherein
a) said isolated oocyte further comprises a zona pellucida, said zona pellucida and said plasma membrane defining a perivitelline space; and, b) contacting said plasma membrane comprises introducing into said perivitelline space said composition.
- 38. A composition comprising an isolated oocyte and an effective concentration of at least a first lentivirus vector comprising a nucleotide sequence of interest, wherein said oocyte is from a vertebrate.
- 39. The composition of claim 38, wherein
a) said isolated oocyte further comprises a zona pellucida, said zona pellucida and said plasma membrane defining a perivitelline space; and, b) said lentivirus vector is in the perivitelline space.
- 40. A method of integrating a nucleotide sequence of interest into a genome of a vertebrate comprising:
a) providing an isolated blastula having a plasma membrane; and, b) contacting said plasma membrane with a composition comprising at least a first lentivirus vector comprising the nucleotide sequence of interest.
- 41. The method of claim 40, wherein the lentivirus vector is derived from a virus selected from the group consisting of a human immunodeficiency virus and a simian immunodeficiency virus.
- 42. The method of claim 40, wherein the lentiviral vector is a trans-viral vector.
- 43. A composition comprising an isolated blastula and an effective concentration of at least a first lentivirus vector comprising a nucleotide sequence of interest, wherein said blastula is from a non-human vertebrate.
CROSS REFERENCE TO RELATED APPLICATION
[0001] This Application claims the benefit of U.S. Provisional Application No. 60/398,476, filed Jul. 25, 2002 and U.S. Provisional Application No. 60/344,841, filed Dec. 21, 2001, both of which are herein incorporated by reference in their entirety.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60398476 |
Jul 2002 |
US |
|
60344841 |
Dec 2001 |
US |