A Plasmid-Based Reverse Genetics System for Animal Double-Stranded RNA Viruses

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

Research output: Contribution to journalArticle

Abstract

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 σ1 and σ3 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.

Original languageEnglish (US)
Pages (from-to)147-157
Number of pages11
JournalCell Host and Microbe
Volume1
Issue number2
DOIs
StatePublished - Apr 19 2007

Keywords

  • MICROBIO

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Virology

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    Kobayashi, T., Antar, A. A. R., Boehme, K. W., Danthi, P., Eby, E. A., Guglielmi, K. M., Holm, G. H., Johnson, E. M., Maginnis, M. S., Naik, S., Skelton, W. B., Wetzel, J. D., Wilson, G. J., Chappell, J. D., & Dermody, T. S. (2007). A Plasmid-Based Reverse Genetics System for Animal Double-Stranded RNA Viruses. Cell Host and Microbe, 1(2), 147-157. https://doi.org/10.1016/j.chom.2007.03.003