Adenoviruses for control of gene expression

• US 6,974,694 B2
• Filed: 10/17/2001
• Issued: 12/13/2005
• Est. Priority Date: 06/07/1995
• Status: Expired due to Fees

• Current Assignee: Advec Incorporated
• Original Assignee: Advec Incorporated
• Inventors: Wang, Ping, Anton, Martina, Rudnicki, Michael A., Bacchetti, Silvia, Muller, William J., Graham, Frank L.
• Primary Examiner(s): McKelvey, Terry
• Assistant Examiner(s): VOGEL, NANCY TREPTOW
• Application Number:
  US09/981,685
• Publication Number:
  US 20030100523A1
• Time in Patent Office: 1,518 Days
• Field of Search: 435/320.1, 435/462, 435/456, 514/44
• US Class Current: 435/320.1
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2710/10343 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2800/30 Vector systems comprising sequences for excision in presence of a recombinase, e.g. loxP or FRT
• CPC Class Codes: C12N 2830/002 inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor

Abstract

The invention claims a class of adenovirus vectors for delivering recombinases to a large number of cells of different origins, and methods for regulating the expression of a gene in transfected mammalian cells in culture and in cells of transgenic animals, comprising infecting said cells with an Ad vector encoding a recombinase whose target site is present at or adjacent to the gene, wherein the action of the recombinase regulates the expression of said gene.

Claims

1. An adenovirus comprising an inserted expression cassette comprising a promoter sequence, a coding sequence of a gene, and a transcription termination site, and site-specific recombinase target sites positioned to remove or invert a portion of the expression cassette, whereby recombination between said target sites mediated by a site-specific recombinase alters expression of the coding sequence of the gene.

2. An adenovirus comprising an inserted expression cassette comprising a promoter sequence, a coding sequence of a gene, and a transcription termination site, and site-specific recombinase target sites flanking the promoter sequence of said expression cassette that promotes expression of the gene, whereby recombination between said target sites mediated by a site-specific recombinase removes the promoter sequence, resulting in decreased expression of the coding sequence of the gene.

3. The adenovirus of claim 2, wherein the coding sequence of the gene is from a non-adenoviral source.

4. An adenovirus comprising an inserted expression cassette comprising a promoter sequence, a coding sequence of a gene, and a transcription termination site, the promoter sequence directed away from said gene, and two site-specific recombinase target sites flanking said promoter sequence but oriented in opposite orientation to one another, whereby recombination between said target sites mediated by a site-specific recombinase inverts the promoter sequence, resulting in increased expression of the coding sequence of the gene.

5. The adenovirus of claim 4, wherein the coding sequence of the gene is from a non-adenoviral source.

6. An adenovirus comprising an inserted expression cassette comprising a promoter sequence, a coding sequence of a gene, a DNA spacer sequence located between the promoter sequence and the coding sequence, and a transcription termination site, and site-specific recombinase target sites flanking the DNA spacer sequence, whereby recombination between said target sites mediated by a site-specific recombinase removes the DNA spacer sequence, resulting in increased expression of the coding sequence of the gene.

7. The adenovirus of claim 6, wherein the coding sequence of the gene is from a non-adenoviral source.

8. An adenovirus comprising an inserted expression cassette comprising a promoter sequence, a coding sequence of a gene, and a transcription termination site, and site-specific recombinase target sites flanking the coding sequence of the gene, whereby recombination between said target sites mediated by a site-specific recombinase removes the coding sequence, resulting in decreased of the coding sequence of the gene.

9. The adenovirus of claim 8, wherein the coding sequence of the gene is from a non-adenoviral source.

10. An adenovirus comprising an inserted expression cassette comprising a promoter sequence, a coding sequence of a gene, and a transcription termination site, a portion of said expression cassette comprising the coding sequence oriented in an opposite direction to normal translation of the coding sequence of the gene, and two site-specific recombinase target sites flanking said coding sequence but oriented in opposite orientation to one another, whereby recombination between said target sites mediated by a site-specific recombinase inverts the coding sequence, resulting in increased expression of the coding sequence of the gene.

11. The adenovirus of claim 10, wherein the coding sequence of the gene is from a non-adenoviral source.

12. An adenovirus comprising a promoter sequence, a coding sequence of a gene, a transcription termination site, and site-specific recombinase target sites flanking the coding sequence of the gene, whereby recombination between said target sites mediated by a site-specific recombinase removes the coding sequence of the gene, resulting in decreased expression of the gene.

13. The adenovirus of claim 12, wherein the coding sequence of the gene is from a non-adenoviral source.

14. An adenovirus comprising a promoter sequence, a coding sequence of a gene, and a transcription termination site, said coding sequence of the gene oriented in an opposite direction to normal translation of the gene, and two site-specific recombinase target sites flanking said coding sequence of the gene but oriented in opposite orientation to one another, whereby recombination between said target sites mediated by a site-specific recombinase inverts the gene, resulting in increased expression of the coding sequence of the gene.

15. The adenovirus of claim 14, wherein the coding sequence of the gene is from a non-adenoviral source.