Attenuated mycobacteria as vectors for gene delivery to mammalian cells
View Patent ↗Provided are mycobacteria comprising a recombinant gene operably liked to a mammalian promoter that directs expression of the recombinant gene from a mammalian cell. Also provided are mammalian cells comprising the above mycobacteria. Additionally provided are mycobacterial plasmids capable of replication in a mycobacterium. Further provided are methods of expressing a recombinant gene in a mammalian cell.
1. A mycobacterium comprising a recombinant gene operably linked to a mammalian promoter that directs expression of the recombinant gene from a mammalian cell, wherein the mycobacterium is M. smegmatis or M. bovis BCG, and wherein the mycobacterium overexpresses (i) RepA, (ii) RepB or (iii) RepA and RepB.
2. The mycobacterium of claim 1 , wherein the mycobacterium is M. bovis BCG.
3. The mycobacterium of claim 1 , wherein the mycobacterium is M. smegmatis.
4. The mycobacterium of claim 1 , wherein the mycobacterium overexpresses RepA.
5. The mycobacterium of claim 1 , wherein the mycobacterium overexpresses RepB.
6. The mycobacterium of claim 1 , wherein the mycobacterium overexpresses RepA and RepB.
7. The mycobacterium of claim 1 , wherein the mycobacterium is capable of hyperconjugation.
8. The mycobacterium of claim 1 , wherein the promoter is a human cytomegalovirus immediate-early promoter.
9. The mycobacterium of claim 1 , wherein the gene encodes a protein.
10. The mycobacterium of claim 9 , wherein the protein is an antigen of a human pathogen.
11. The mycobacterium of claim 10 , wherein the human pathogen is influenza.
12. The mycobacterium of claim 1 , wherein the gene is on a mycobacterial plasmid.
13. A mammalian cell comprising the recombinant mycobacterium of claim 1 .
14. The cell of claim 13 , wherein the mycobacterium is M. bovis BCG.
15. The cell of claim 13 , wherein the mycobacterium is M. smegmatis.
16. The cell of claim 13 , wherein the mycobacterium overexpresses RepA.
17. The cell of claim 13 , wherein the mycobacterium overexpresses RepB.
18. The cell of claim 13 , wherein the mycobacterium overexpresses RepA and RepB.
19. A mycobacterial plasmid capable of replication in a mycobacterium, wherein the plasmid comprises a recombinant gene operably liked to a mammalian promoter that directs expression of the recombinant gene from a mammalian cell, wherein the mycobacterium is M. smegmatis or M. bovis BCG, and wherein the mycobacterial plasmid directs overexpression of (i) RepA, (ii) RepB or (iii) RepA and RepB.
20. The plasmid of claim 19 , wherein the mycobacterium is M. bovis BCG.
21. The plasmid of claim 19 , wherein the mycobacterium is M. smegmatis.
22. The plasmid of claim 19 , which directs overexpression of RepA.
23. The plasmid of claim 19 , which directs overexpression of RepB.
24. The plasmid of claim 19 , which directs overexpression of RepA and RepB.
25. A method of expressing a recombinant gene in a mammalian cell, the method comprising infecting the cell with the mycobacterium of claim 1 .
26. The method of claim 25 , wherein the mycobacterium is M. bovis BCG.
27. The method of claim 25 , wherein the mycobacterium is M. smegmatis.
28. The method of claim 25 , wherein the mycobacterium overexpresses RepA.
29. The method of claim 25 , wherein the mycobacterium overexpresses RepB.
30. The method of claim 25 , wherein the mycobacterium overexpresses RepA and RepB.
31. The method of claim 25 , wherein the gene encodes an antigen of a human pathogen.