DOUBLE-STRANDED/SELF-COMPLEMENTARY VECTORS WITH A TRUNCATED CBA PROMOTER AND METHODS OF GENE DELIVERY

• US 20100069467A1
• Filed: 04/27/2007
• Published: 03/18/2010
• Est. Priority Date: 04/28/2006
• Status: Active Grant

• Current Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Original Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Inventors: Boye, Sanford L., Byrne, Barry Jonh, Hauswirth, William W.
• Granted Patent: US 8,298,818 B2
• Time in Patent Office: Days
• US Class Current: 514/44.R
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2830/15 chimeric enhancer/promoter combination

Abstract

The present inventors concerns vectors carrying a truncated chimeric CMV-chicken β-actin (smCBA) promoter in which the hybrid chicken β-actin/rabbit β-globin intron is greatly shortened, and their use to deliver to an operatively linked polynucleotide to host cells in vitro or in vivo, resulting in expression of the polynucleotide in the host cells. In one embodiment, the vector carrying the smCBA promoter is administered to the eye. In another embodiment, the vector carrying the smCBA promoter is a self-complementary adeno-associated virus (AAV). The AAV vector may be of any serotype (e.g., type 1, type 2, type 3, type 4, type 5, type 6, type 7, type 8, type 9, type 10). In another embodiment, a self-complementary vector carrying the smCBA promoter is administered to the eye. Another aspect of the invention concerns host cells carrying a vector of the invention. Another aspect of the invention concerns pharmaceutical composition comprising the vectors or host cells of the invention, and a pharmaceutically acceptable carrier.

Claims

1. A vector comprising a truncated chimeric CMV-chicken β

-actin promoter.

2. The vector of claim 1, wherein said truncated chimeric CMV-chicken β

-actin promoter consists of the nucleic acid sequence of SEQ ID NO;

1.

3. The vector of claim 1, further comprising a heterologous polynucleotide operably linked to the truncated chimeric CMV-chicken β

-actin promoter.

4. The vector of claim 3, wherein the polynucleotide encodes a polypeptide, an interfering RNA molecule or an antisense oligonucleotide.

6. The vector of claim 1, wherein the vector is a viral vector.

7. The vector of claim 1, wherein the vector is a self-complementary adeno-associated virus (scAAV).

5. (canceled)

8. An isolated host cell comprising a vector comprising a truncated chimeric CMV-chicken β

-actin promoter.

9. The isolated cell of claim 8, wherein the cell is a human cell or a non-human animal cell.

11. The isolated cell of claim 8, wherein the cell is of a type selected from the group consisting of brain cell, liver cell, lung cell, cardiac cell, ocular cell, and smooth muscle cell.

12. The isolated cell of claim 8, wherein the cell is a neuron.

10. (canceled)

13. (canceled)

14. A method for delivering a polynucleotide to a cell, comprising administering an effective amount of a vector comprising a truncated chimeric CMV-chicken β

-actin promoter to the cell, wherein the polynucleotide is expressed in the cell.

15. The method of claim 14, wherein said administering is carried out in vitro or in vivo.

17. The method of claim 14, wherein the cell is a human cell or a non-human animal cell.

19. The method of claim 14, wherein the cell is of a type selected from the group consisting of brain cell, liver cell, lung cell, cardiac cell, ocular cell, and smooth muscle cell.

20. The method of claim 14, wherein the cell is a neuron.

21. The method of claim 14, wherein said administering comprising administering an effective amount of the vector to a human or animal systemically or administering an effective amount of the vector to a human or animal locally at the site of the cell.

23. The method of claim 14, wherein the vector is administered to cells of the eye.

24. The method of claim 23, wherein the vector is administered to the vitreous or subretinally.

25. The method of claim 14, wherein the polynucleotide is heterologous to the cell or native to the cell.

27. The method of claim 14, wherein the polynucleotide encodes a polypeptide, an interfering RNA molecule or an antisense oligonucleotide.

16. (canceled)

18. (canceled)

22. (canceled)

26. (canceled)

28-29. -29. (canceled)