A plasmid-based reverse genetics system for animal double-stranded RNA viruses.

Kobayashi T, Antar AA, Boehme KW, Danthi P, Eby EA, Guglielmi KM, Holm GH, Johnson EM, Maginnis MS, Naik S, Skelton WB, Wetzel JD, Wilson GJ, Chappell JD, Dermody TS
Cell Host Microbe. 2007 1 (2): 147-57

PMID: 18005692 · PMCID: PMC2034303 · DOI:10.1016/j.chom.2007.03.003

Mammalian orthoreoviruses (reoviruses) are highly tractable experimental models for studies of double-stranded (ds) RNA virus replication and pathogenesis. Reoviruses infect respiratory and intestinal epithelium and disseminate systemically in newborn animals. Until now, a strategy to rescue infectious virus from cloned cDNA has not been available for any member of the Reoviridae family of dsRNA viruses. We report the generation of viable reovirus following plasmid transfection of murine L929 (L) cells using a strategy free of helper virus and independent of selection. We used the reovirus reverse genetics system to introduce mutations into viral capsid proteins sigma1 and sigma3 and to rescue a virus that expresses a green fluorescent protein (GFP) transgene, thus demonstrating the tractability of this technology. The plasmid-based reverse genetics approach described here can be exploited for studies of reovirus replication and pathogenesis and used to develop reovirus as a vaccine vector.

MeSH Terms (13)

Animals Disease Models, Animal Genome, Viral Mammals Plasmids Recombination, Genetic Reoviridae Reoviridae Infections RNA, Double-Stranded RNA Viruses Transfection Vaccines, Synthetic Viral Vaccines

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