Reverse Genetics for Mammalian Orthoreovirus

Methods Mol Biol. 2017:1602:1-10. doi: 10.1007/978-1-4939-6964-7_1.

Abstract

Reverse genetics allows introduction of specific alterations into a viral genome. Studies performed with mutant viruses generated using reverse genetics approaches have contributed immeasurably to our understanding of viral replication and pathogenesis, and also have led to development of novel vaccines and virus-based vectors. Here, we describe the reverse genetics system that allows for production and recovery of mammalian orthoreovirus, a double-stranded (ds) RNA virus, from plasmids that encode the viral genome.

Keywords: Double-stranded RNA virus; Plasmid-based reverse genetics; Recombinant virus; Reovirus; T7 RNA polymerase; Viral reassortment.

MeSH terms

  • Animals
  • Cell Line
  • Genes, Viral
  • Genome, Viral
  • Humans
  • Mice
  • Mutation
  • Orthoreovirus, Mammalian / genetics*
  • Plasmids / genetics
  • RNA, Double-Stranded
  • RNA, Viral
  • Reassortant Viruses / genetics
  • Recombination, Genetic
  • Reverse Genetics* / methods

Substances

  • RNA, Double-Stranded
  • RNA, Viral