Bovine immunodeficiency virus (BIV) based vectors
BIV packaging constructs, BIV packaging cell lines, methods of making BIV packaging cells and methods of making BIV producer cells are described.
1. A bovine immunodeficiency virus (BIV) packaging construct comprising a BIV DNA sequence fragment comprising at least a gag or a pol gene of BIV, and a promoter operably linked to said BIV DNA fragment.
2. The BIV construct of claim 1 further comprising an internal ribosome binding site.
3. The BIV construct of claim 1 further comprising a heterologous intron.
4. A BIV packaging cell line comprising an established cell line transformed by the BIV construct of claim 1 .
5. A packaging construct comprising a promoter operatively linked to a BIV gag/pol coding sequence and a polyadenylation signal at the 3′ end of said gag/pol coding sequence.
6. The packaging construct of claim 5 , further comprising an intron upstream of said gag/pol coding sequence.
7. The packaging construct of claim 5 , further comprising an RNA transport element.
8. The packaging construct of claim 7 , wherein said RNA transport element comprises a lentiviral rev response element (RRE).
9. The packaging construct of claim 8 , wherein said lentiviral RRE comprises a BIV RRE.
10. The packaging construct of claim 7 , wherein said RNA transport element comprises a constitutive transport element (CTE).
11. The packaging construct of claim 10 , wherein said CTE is a Mason-Pfizer monkey virus CTE.
12. The packaging construct of claim 8 , further comprising a Rev coding sequence.
13. A method of making a packaging cell comprising the steps of (1) transfecting a cell with a BIV gag/pol coding sequence and (2) transfecting the cell with a viral envelope coding sequence.
14. The method of claim 13 , wherein said cell is a mammalian cell.
15. The method of claim 14 , wherein said mammalian cell is a human cell.
16. A method of making a producer cell comprising transfecting the packaging cell of claim 13 with a transfer vector comprising a transgene.