title: Rescue and production of vaccine and therapeutic adenovirus vectors expressing inhibitory transgenes.
abstract: Expression of certain transgenes from an adenovirus vector can be deleterious to its own replication. This can result in the inhibition of virus rescue, reduced viral yields, or, in the worst case, make it impossible to construct a vector expressing the inhibiting transgene product. A gene regulation system based on the tet operon was used to allow the rescue and efficient growth of adenovectors that express transgenes to high levels. A key advantage to this system is that repression of transgene expression is mediated by the packaging cell line, thus, expression of regulatory products from the adenovector are not required. This provides a simple, broadly applicable system wherein transgene repression is constitutive during vector rescue and growth and there is no effect on adenovector-mediated expression of gene products in transduced cells. Several high-level expression vectors based on first- and second-generation adenovectors were rescued and produced to high titer that otherwise could not be grown. Yields of adenovectors expressing inhibitory transgene products were increased, and the overgrowth of cultures by adenovectors with nonfunctional expression cassettes was prevented. The gene regulation system is a significant advancement for the development of adenovirus vectors for vaccine and other gene transfer applications.
authors.affiliation: GenVec, Inc. 65 West Watkins Mill Rd, Gaithersburg MD 20878, USA.
authors.forename: Jason G D
authors.initials: JG
authors.lastname: Gall
authors.forename: Alena
authors.initials: A
authors.lastname: Lizonova
authors.forename: Damodar
authors.initials: D
authors.lastname: EttyReddy
authors.forename: Duncan
authors.initials: D
authors.lastname: McVey
authors.forename: Mohammed
authors.initials: M
authors.lastname: Zuber
authors.forename: Imre
authors.initials: I
authors.lastname: Kovesdi
authors.forename: Barbara
authors.initials: B
authors.lastname: Aughtman
authors.forename: C Richter
authors.initials: CR
authors.lastname: King
authors.forename: Douglas E
authors.initials: DE
authors.lastname: Brough
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005822:Genetic Vectors; D006801:Humans; D019076:Transgenes; D014765:Viral Vaccines / Q000235:genetics*; D014779:Virus Replication
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D014765:Viral Vaccines