Description of Invention:
This invention relates to a VAC-BAC shuttle vector system for the creation of recombinant poxviruses from DNA cloned in a bacterial artificial chromosome. A VAC-BAC is a bacterial artificial chromosome (BAC) containing a vaccinia virus genome (VAC) that can replicate in bacteria and produce infectious virus in mammalian cells.
The following are some of the uses for a VAC-BAC:
VAC-BACs can be used to modify vaccinia virus DNA by deletion, insertion or point mutation or add new DNA to the VAC genome with methods developed for bacterial plasmids, rather than by recombination in mammalian cells.
It can be used to produce recombinant vaccinia viruses for gene expression.
It can be used for the production of modified vaccinia viruses that have improved safety or immunogenicity.
Advantages of the VAC-BAC shuttle system:
VAC-BACs are clonally purified from bacterial colonies before virus reconstitution in mammalian cells.
Manipulation of DNA is much simpler and faster in bacteria than in mammalian cells.
Modified genomes can be characterized prior to virus reconstitution.
Only virus with modified genomes will be produced so that virus plaque isolations are not needed.
Generation of a stock of virus from a VAC-BAC is accomplished within a week rather than many weeks.
Multiple viruses can be generated at the same time since plaque purification is unnecessary.
DHHS Reference No. E-355-2001/1 --
U.S. Provisional Application No. 60/402,824 filed 09 Aug 2002
DHHS Reference No. E-355-2001/2 --
International Patent Application No. PCT/US03/11183 filed 10 Apr 2003, which published as WO 03/087330 A2 on 23 Oct 2003
U.S. Patent Application No. 10/959,392 filed 05 Oct 2004
European Patent Application No. 037183431 filed 10 Apr 2003
Relevant Publication:
A Domi and B Moss. Cloning the vaccinia virus genome as a bacterial artificial chromosome in Escherichia coli and recovery of infectious virus in mammalian cells. Proc Natl Acad Sci USA. 2002 Sep 17;99(19):12415-12420. [PubMed abs]
A Domi and B Moss. Engineering of a vaccinia virus bacterial artificial chromosome in Escherichia coli by bacteriophage lambda-based recombination. Nat Methods. 2005 Feb;2(2):95-97. [PubMed abs]
Licensing Status: In addition to licensing, the technology is available for further development through collaborative research opportunities with the inventors.
