Claims
- 1. A method for producing a recombinant adenovirus comprising a gene of interest, said method not producing replication competent adenovirus having a functional E1 region, said method comprising:
providing a cell, said cell harboring a first nucleic acid comprising adenoviral nucleic acid; transferring recombinant nucleic acid into said cell, said recombinant nucleic acid comprising:
a second nucleic acid containing adenoviral nucleic acid including at least one encapsidating signal, and at least one functional Inverted Terminal Repeat, said recombinant nucleic acid lacking overlapping sequences with the cellular nucleic acid having a functional E1 region; culturing said cell; and harvesting the recombinant adenovirus produced from said cell.
- 2. The method according to claim 1 wherein said recombinant nucleic acid is in linear form and comprises functional Inverted Terminal Repeats at or near both termini.
- 3. The method according to claim 1 wherein said cell is a primary cell.
- 4. The method according to claim 1 wherein said recombinant nucleic acid is DNA.
- 5. A method ofproducing, in a producer cell, recombinant adenovirus comprising a gene of interest, said method comprising:
culturing, in a suitable medium, a producer cell comprising one or more recombinant nucleic acid molecules having no overlapping sequences with respect to one another, wherein said producer cell expresses at least adenoviral E1A region gene products; and harvesting recombinant adenovirus produced from said cell.
- 6. The method according to claim 5 wherein one or more of said recombinant nucleic acid molecules of said producer cell further has a mutated E2A region of an adenovirus of the family Adenoviridae.
- 7. A method of producing a recombinant adenovirus comprising a gene of interest, said method comprising:
culturing a producer cell in a suitable medium and harvesting said adenovirus therefrom, wherein said producer cell comprises:
one or more recombinant nucleic acid molecules having no overlapping sequences with respect to one another, and wherein said producer cell contains a gene encoding for adenoviral E1 and E2A region gene products; and harvesting recombinant adenovirus produced from said producer cell.
- 8. The method according to claim 7 wherein the gene encoding for the adenoviral E2A region gene products is under the control of an inducible promoter.
- 9. The method according to claim 7 wherein the gene encoding for the E2A region gene products is mutated so that at least one of its products is temperature sensitive.
- 10. The method according to claim 8 wherein the gene encoding for the E2A region gene products is mutated so that at least one of its products is temperature sensitive.
- 11. The method according to claim 8 wherein said producer cell is of monkey origin.
- 12. The method according to claim 11 wherein said producer cell is of monkey origin.
- 13. A method according to claim 7 wherein one or more of said recombinant nucleic acid molecules of said producer cell further has a mutated E2A region of an adenovirus of the family Adenoviridae.
- 14. A method of producing a recombinant adenovirus comprising a gene of interest, said method comprising:
culturing a producer cell in a suitable medium, said producer cell comprising:
one or more recombinant nucleic acid molecules having no overlapping sequences with respect to one another, said producer cell further expressing adenoviral E1 and E2A region gene products, wherein said E2A region is mutated so that at least one of its products is temperature sensitive; and harvesting said recombinant adenovirus from said cell.
- 15. A method of producing a recombinant adenovirus comprising a gene of interest, said method comprising:
culturing a producer cell in a suitable medium, said producer cell comprising:
one or more recombinant nucleic acid molecules having no overlapping sequences with respect to one another, and DNA sequences encoding the adenoviral E1A and E1B region gene products; and harvesting recombinant adenovirus from said cell.
- 16. The method according to claim 15 wherein said recombinant nucleic acid molecule further comprises DNA sequences encoding adenoviral E2A region gene products.
- 17. The method according to claim 16 wherein one of said DNA sequences encoding the E2A region gene product is selected from the group consisting of a DNA sequence encoding the wild-type E2A region operably linked to an inducible promoter and a DNA sequence encoding a temperature sensitive 125 mutation.
Priority Claims (2)
Number |
Date |
Country |
Kind |
EP 95201611.1 |
Jun 1995 |
EP |
|
EP 95201728.3 |
Jun 1995 |
EP |
|
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of co-pending, co-owned U.S. patent application Ser. No. 09/356,575, filed on Jul. 19, 1999, U.S. Pat. No. 6,265,212, the contents of which are incorporated by this reference, which is a continuation-in-part of U.S. application Ser. No. 08/793,170 filed Mar. 25, 1997, pending, incorporated herein by reference, which is the national stage filing of PCT/NL96/00244 filed Jun. 14, 1996.
Divisions (1)
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Number |
Date |
Country |
Parent |
09356575 |
Jul 1999 |
US |
Child |
09912552 |
Jul 2001 |
US |
Continuation in Parts (1)
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Number |
Date |
Country |
Parent |
08793170 |
Mar 1997 |
US |
Child |
09356575 |
Jul 1999 |
US |