J Gen Virol. 2013 Jan 30. [Epub ahead of print]
Arenavirus reverse genetics for vaccine development.
Source
University of Rochester;
Abstract
Arenaviruses are important human pathogens with no FDA-licensed vaccines available and current antiviral therapy being limited to an off-label use of the nucleoside analog ribavirin of limited prophylactic efficacy. The development of reverse genetics systems represented a major breakthrough in the arenavirus research. However, rescue of recombinant arenaviruses using current reverse genetics systems has been restricted to rodent cells. In this study we describe the rescue of recombinant arenaviruses from human 293T cells and Vero cells, a FDA-approved line for vaccine development. We also describe the generation of novel vectors that mediate synthesis of both negative-sense genome RNA and positive-sense mRNA species of LCMV directed by the human RNA polymerases I and II, respectively, within the same plasmid. This approach reduces to half the number of vectors required for arenavirus rescue, which could facilitate virus rescue in cell lines approved for human vaccine production but that cannot be transfected at high efficiencies. We have shown the feasibility of this approach by rescuing both the Old World prototypic arenavirus LCMV and the live attenuated vaccine Candid#1 strain of the New World arenavirus Junin. Moreover, we show the feasibility of using these novel strategies for efficient rescue of recombinant tri-segmented both LCMV and Candid#1.
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