Transgenic, non-human animals containing a coxsackie/adenovirus receptor (CAR)

• US 20020016974A1
• Filed: 02/16/2001
• Published: 02/07/2002
• Est. Priority Date: 02/16/2000
• Status: Abandoned Application

• Current Assignee: University Of Massachusetts
• Original Assignee: University Of Massachusetts
• Inventors: Schmidt, Madelyn R., Woodland, Robert T., Piekos, Brian
• Application Number:
  US09/788,025
• Publication Number:
  US 20020016974A1
• Time in Patent Office: Days
• US Class Current: 800/4
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 2267/0331 Animal model for proliferative diseases
• CPC Class Codes: A01K 2267/0337 Animal models for infectious diseases
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination

Abstract

The invention features a transgenic animal, which expresses a Coxsackie-adenovirus receptor (CAR) in its cells. The invention provides a new animal model for effecting adenovirus-mediated gene delivery and for testing gene function in vivo, and also provides new opportunities to assess gene function in vitro using freshly isolated cells.

Claims

1. A transgenic, non-human animal, one or more of whose cells comprise a transgene comprising a regulatory sequence operably linked to a nucleic acid sequence encoding a Coxsackie/adenovirus receptor (CAR), wherein the transgene is expressed in one or more cells of the transgenic animal resulting in the animal exhibiting increased susceptibility to adenovirus infection as compared to a wildtype animal.

2. The animal of claim 1, wherein the transgene is expressed in one or more hematopoietic cells.

3. The animal of claim 1, wherein the transgene is expressed in one or more lymphocytes.

4. The animal of claim 1, wherein the animal develops or has a disorder.

5. The animal of claim 1, wherein the animal has an immunodeficiency disorder.

6. The animal of claim 5, wherein the immunodeficiency disorder is X-linked severe combined immune deficiency, X-linked agammaglobulinemia, or Wiskott-Aldrich syndrome.

7. The animal of claim 1, wherein the regulatory sequence is constitutive or inducible.

8. The animal of claim 1, wherein the regulatory sequence is an MHC class I promoter.

9. The animal of claim 1, wherein the regulatory sequence is an exogenous sequence.

10. The animal of claim 9, wherein the animal comprises one or more cells that comprise an endogenous regulatory sequence that controls the transcription of an endogenous CAR, and wherein the exogenous regulatory sequence controlling transcription of a CAR is different from the endogenous regulatory sequence controlling the transcription of the endogenous CAR.

11. The animal of claim 1, wherein the nucleic acid sequence is substantially identical to a nucleic acid sequence encoding a mouse or human CAR.

12. The animal of claim 1, wherein the animal is a rodent.

13. The animal of claim 1, wherein the animal is a mouse.

15. The nucleic acid sequence of claim 14, wherein the nucleic acid sequence encodes a human or mouse CAR.

16. A vector comprising the nucleic acid sequence of claim 14.

17. The vector of claim 16, wherein the vector is a plasmid or a viral vector.

18. A cell containing the nucleic acid sequence of claim 14.

19. A cell derived from the animal of claim 1.

20. The cell of claim 19, wherein the cell is a lymphocyte, a liver cell, or a spleen cell.

24. A method of determining whether an adenoviral vector infects a cell in vivo, the method comprising:

(a) contacting a cell from the transgenic animal of claim 1 in vivo with an adenoviral vector comprising a test gene encoding a test protein; and

(b) determining expression of the test gene in the cell, wherein expression of the test gene in the cell indicates that the cell can be infected with the adenoviral vector.

14. An isolated nucleic acid molecule comprising an MHC class 1 regulatory sequence operably linked to a nucleic acid sequence that encodes a CAR.

21. A method of infecting a cell with an adenovirus, the method comprising contacting a cell engineered to express a nucleic acid sequence encoding a Coxsackie-adenovirus receptor (CAR) with an adenovirus such that the adenovirus infects the cell.

22. The method of claim 21, wherein the cell is normally refractory to adenovirus infection.

23. The method of claims 21, wherein the cell is in an animal.

25. The method of claim 24, wherein the cell is normally refractory to adenoviral vector infection.

27. The method of claim 26, wherein the cell is a tumor cell.

28. The method of claim 26, wherein the tumor cell is a prostate tumor cell.

26. A method of testing the efficiency of a regulatory sequence to express a nucleic acid sequence in a cell in vivo, the method comprising:

(a) contacting a cell engineered to express a nucleic acid sequence encoding a Coxsackie/adenovirus receptor (CAR) in vivo with an adenoviral vector comprising a test regulatory sequence operably linked to a reporter gene, and

(b) determining the level of expression of the reporter gene in the cell, wherein the level of expression is an indication of the efficiency of the regulatory sequence.