Development of an entirely plasmid-based reverse genetics system for 12-segmented double-stranded RNA viruses

Abstract

Significance Coltiviruses, including important pathogens such as Colorado tick fever virus in humans, have a genome comprising 12 segments of double-stranded (ds) RNA. However, due to the lack of a reverse genetics system, the molecular mechanisms of coltivirus biology are largely unknown. Here, we established an entirely plasmid-based reverse genetics system for Tarumizu tick virus, a member of the genus Coltivirus , isolated from the brain of a dead raccoon dog. Recombinant wild-type and mutant viruses were generated from cloned complementary DNAs. This is a report of a reverse genetics system for Reoviridae viruses harboring a genome comprising 12 segments of dsRNA. This technology provides opportunities for understanding the mechanisms of replication and pathogenesis of viruses within the genus Coltivirus .