Bovine immunodeficiency virus (BIV) based vectors
This invention pertains to BIV constructs encompassing BIV combination vectors, BIV vectors and BIV packaging vectors and particularly the invention pertains to a three vector system comprising: a) a BIV vector construct including a DNA segment from a BIV genome, a packaging sequence to package RNA into virions; a promoter operably linked to the DNA segment; and a transgene operably linked to a second promoter; b) a BIV packaging vector construct comprising a BIV DNA sequence fragment comprising at least a gag gene or pol gene of BIV; a promoter operably linked to the BIV DNA fragment; and a polyadenylation sequence located downstream of the BIV DNA fragment; and c) an expression vector construct comprising a gene encoding a viral surface protein. Also provided is a method for transferring a gene of interest into a mammalian cell.
1. A virion comprising:
(a) at least a portion of a BIV gag coding sequence or a portion of a BIV pol coding sequence;
(b) a viral envelope coding sequence; and
(c) a vector construct comprising:
1) a packaging sequence to package RNA into virions; and
2) a transgene operably linked to a promoter,
wherein at least one of vpw, vpy and tat is deleted from said construct.
2. A virion comprising an RNA vector, said RNA vector comprising a construct comprising a BIV packaging sequence, a transgene operably linked to a promoter, wherein at least one of vpw, vpy and tat is deleted from said vector.
3. The virion of claim 1 , wherein the packaging sequence is a BIV packaging sequence.
4. The virion of claim 1 , wherein said promoter is an LTR promoter.
5. The virion of claim 1 , wherein the transgene is operably linked to a CMV promoter, a PGK promoter or an MND promoter.
6. The virion of claim 1 , wherein said construct further comprises a rev-response element (RRE).
7. The virion of claim 6 , wherein said RRE is a BIV RRE.
8. The virion of claim 1 , wherein said construct further comprises a scaffold attachment region (SAR).
9. The virion of claim 8 , wherein said SAR comprises a human interferon-β SAR.
10. The virion of claim 1 , wherein said construct further comprises a central polypurine tract (cPPT).
11. The virion of claim 1 , wherein said construct further comprises a portion of a U3 region of an LTR.
12. The virion of claim 11 , wherein a portion of the U3 region is deleted or replaced by a heterologous sequence.
13. The virion of claim 1 , wherein the transgene is operably linked to an internal promoter.
14. The virion of claim 11 , wherein one or more nucleotides in the U3 region are mutated or deleted to diminish or eliminate U3-mediated transcription.
15. The virion of claim 1 , wherein any start codons in the packaging sequence be eliminated by deletion or mutation.
16. The virion of claim 11 , wherein the U3 element further comprises a sequence that enhances polyadenylation.
17. The virion of claim 1 , wherein said construct further comprises a splice donor site.
18. The virion of claim 10 , wherein the cPPT is a BIV cPPT.
19. The virion of claim 1 , wherein said construct further comprises a 3′ polypurine tract.
20. The virion of claim 1 , wherein said construct further comprises an RNA transport element.
21. The virion of claim 20 , wherein the RNA transport element is a constitutive transport element (CTE).
22. The virion of claim 21 , wherein the CTE is a Mason-Pfizer Monkey Virus CTE.
23. The virion of claim 2 , wherein said promoter is an LTR promoter.
24. The virion of claim 2 , wherein the transgene is operably linked to a CMV promoter, a PGK promoter or an MND promoter.
25. The virion of claim 2 , wherein said vector further comprises a rev-response element (RRE).
26. The virion of claim 25 , wherein said RRE is a BIV RRE.
27. The virion of claim 2 , wherein said vector further comprises a scaffold attachment region (SAR).
28. The virion of claim 27 , wherein said SAR comprises a human interferon-β SAR.
29. The virion of claim 2 , wherein said vector further comprises a central polypurine tract (cPPT).
30. The virion of claim 2 , wherein said vector further comprises a portion of a U3 region of an LTR.
31. The virion of claim 30 , wherein a portion of the U3 region is deleted or replaced by a heterologous sequence.
32. The virion of claim 2 , wherein the transgene is operably linked to an internal promoter.
33. The virion of claim 30 , wherein one or more nucleotides in the U3 region are mutated or deleted to diminish or eliminate U3-mediated transcription.
34. The virion of claim 2 , wherein any start codons in the packaging sequence be eliminated by deletion or mutation.
35. The virion of claim 30 , wherein the U3 element further comprises a sequence that enhances polyadenylation.
36. The virion of claim 2 , wherein said vector further comprises a splice donor site.
37. The virion of claim 29 , wherein the cPPT is a BIV cPPT.
38. The virion of claim 2 , wherein said vector further comprises a 3′ polypurine tract.
39. The virion of claim 2 , wherein said vector further comprises an RNA transport element.
40. The virion of claim 39 , wherein the RNA transport element is a constitutive transport element (CTE).
41. The virion of claim 40 , wherein the GTE is a Mason-Pfizer Monkey Virus CTE.
42. The virion of claim 1 , further comprising a rev coding sequence.
43. The virion of claim 2 , further comprising a rev coding sequence.
44. The virion of claim 1 , wherein vpw is deleted from said construct.
45. The virion of claim 1 , wherein vpy is deleted from said construct.
46. The virion of claim 1 , wherein tat is deleted from said construct.
47. The virion of claim 2 , wherein vpw is deleted from said construct.
48. The virion of claim 2 , wherein vpy is deleted from said construct.
49. The virion of claim 2 , wherein tat is deleted from said construct.