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 produced by a producer cell comprising:
a) a BIV gag/pol coding sequence;
b) a viral envelope coding sequence; and
c) a vector construct comprising a promoter linked to a first BIV R region, a BIV U5 element linked to the first BIV R region, a packaging sequence, a transgene, a 3′ polypurine tract, and a BIV U3 element linked to a second BIV R region, wherein the promoter initiates RNA transcription of the vector construct, and 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 first BIV R region linked to a BIV U5 element, a packaging sequence, a transgene operably linked to a promoter, a 3′ polypurine tract, and a BIV U3 element linked to a second BIV R region, and wherein at least one of vpw, vpy and tat is deleted from said vector.
3. A virion according to claim 1 , wherein the transgene is operably linked to an internal promoter.
4. A virion according to claim 1 , wherein one or more nucleotide sequences in the U3 element are mutated or deleted in order to diminish or eliminate U3-mediated transcription.
5. A virion according to claim 1 , wherein the packaging sequence is a BIV packaging sequence.
6. A virion according to claim 1 , wherein any start codons in the packaging sequence are eliminated by deletion or mutation.
7. A virion according to claim 1 , wherein the U3 element further comprises a sequence that enhances polyadenylation.
8. A virion according to claim 1 , wherein the major splice donor site has been inactivated or eliminated.
9. A virion according to claim 1 , further comprising a cPPT.
10. A virion according to claim 9 , wherein the cPPT is a BIV cPPT.
11. A virion according to claim 1 , further comprising an RNA transport element.
12. A virion according to claim 11 , wherein the RNA transport element is a lentiviral rev response element (RRE).
13. A virion according to claim 12 , wherein the lentiviral RRE is a BIV RRE.
14. A virion according to claim 11 , wherein the RNA transport element is a constitutive transport element (CTE).
15. A virion according to claim 14 , wherein the CTE is a Mason-Pfizer Monkey Virus CTE.
16. A virion according to claim 2 , wherein the transgene is operably linked to an internal promoter.
17. A virion according to claim 2 , wherein one or more nucleotide sequences in the U3 element are mutated or deleted in order to diminish or eliminate U3-mediated transcription.
18. A virion according to claim 2 , wherein the packaging sequence is a BIV packaging sequence.
19. A virion according to claim 2 , wherein any start codons in the packaging sequence are eliminated by deletion or mutation.
20. A virion according to claim 2 , wherein the U3 element further comprises a sequence that enhances polyadenylation.
21. A virion according to claim 2 , wherein the major splice donor site has been inactivated or eliminated.
22. A virion according to claim 2 , further comprising a cPPT.
23. A virion according to claim 22 , wherein the cPPT is a BIV cPPT.
24. A virion according to claim 2 , further comprising an RNA transport element.
25. A virion according to claim 24 , wherein the RNA transport element is a lentiviral rev response element (RRE).
26. A virion according to claim 25 , wherein the lentiviral RRE is a BIV RRE.
27. A virion according to claim 24 , wherein the RNA transport element is a constitutive transport element (CTE).
28. A virion according to claim 27 , wherein the CTE is a Mason-Pfizer Monkey Virus CTE.