Rescue and production of vaccine and therapeutic adenovirus vectors expressing inhibitory transgenes |
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Authors: | Jason G D Gall Alena Lizonova Damodar EttyReddy Duncan McVey Mohammed Zuber Imre Kovesdi Barbara Aughtman C Richter King Douglas E Brough |
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Institution: | (1) GenVec, Inc, 65 West Watkins Mill Rd, 20878 Gaithersburg, MD, USA;(2) KILA Consultants LLC, 7713 Warbler Lane, 20855 Rockville, MD, USA |
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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. |
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Keywords: | Adenoviruses vaccines gene therapy genetic vectors repressor proteins tet repressor |
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