Adenovirus gene therapy vehicle and cell line

• US 6,281,010 B1
• Filed: 10/27/1995
• Issued: 08/28/2001
• Est. Priority Date: 06/05/1995
• Status: Expired due to Fees

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Wilson, James M., Gao, Guang-Ping
• Primary Examiner(s): Nguyen, Dave Trong
• Application Number:
  US08/549,489
• Time in Patent Office: 2,132 Days
• Field of Search: 514/44, 435/173.3, 435/320.1, 435/69.1, 435/325, 435/62, 435/455, 435/91.4, 424/93.2, 424/93.21
• US Class Current: 435/325
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• 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 2750/14143 viral genome or elements thereof as genetic vector
• CPC Class Codes: Y10S 977/802 Virus-based particle
• CPC Class Codes: Y10S 977/907 Liposome

Abstract

A novel adenovirus E1/E4 expressing packaging cell line is provided, which permits the generation of recombinant adenoviruses deleted in both gene regions. The E1/E4 deleted recombinant adenovirus is capable of expressing a selected transgene product in cells in vivo or in vitro. This recombinant virus is useful in the treatment of genetic disorders.

Claims

1. The cell line 293-10-3 ATCC Accession No. PTA-2361.

2. A method for producing a recombinant adenovirus deleted in the E1 and E4 genes, comprising the step of:

introducing to cell line 293-10-3 ATCC Accession No. PTA-2361

(i) a recombinant vector comprising adenovirus 5′ and

3′

cis elements necessary for replication and packaging flanking a transgene under the control of regulatory sequences that direct expression of said transgene in a host cell, and

(ii) a helper virus providing any adenovirus genes not present in the cell line or in said vector, that are necessary to permit production of said recombinant adenovirus.

3. The method according to claim 2, further comprising culturing said cells under conditions which permit packaging of said transgene into a recombinant E1, E4-deleted adenovirus.

4. The method according to claim 3, further comprising purifying said recombinant adenovirus from cellular debris.

5. The cell line 293-27-18 ATCC Accession No. PTA-2511.

6. A method for producing a recombinant adenovirus deleted in the E1 and E4 genes, comprising the step of:

introducing to cell line 293-27-18 ATCC Accession No. PTA-2511

(i) a recombinant vector comprising adenovirus 5′ and

3′

cis elements necessary for replication and packaging flanking a transgene under the control of regulatory sequences that direct expression of said transgene in a host cell, and

(ii) a helper virus providing any adenovirus genes not present in the cell line or in said vector, that are necessary to permit production of said recombinant adenovirus.

7. The method according to claim 6, further comprising culturing said cells under conditions which permit packaging of said transgene into a recombinant E1, E4-deleted adenovirus.

8. The method according to claim 6, further comprising purifying said recombinant adenovirus from cellular debris.