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

作者：Kobayashi Takeshi; Antar Annukka A R; Boehme Karl W; Danthi Pranav; Eby Elizabeth A; Guglielmi Kristen M; Holm Geoffrey H; Johnson Elizabeth M; Maginnis Melissa S; Naik Sam; Skelton Wesley B; Wetzel J Denise; Wilson Gregory J; Chappell James D^*; Dermody Terence S

来源：Cell Host & Microbe, 2007, 1(2): 147-157.

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 all and sigma 3 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.

• 出版日期2007-4