Development of an entirely plasmid-based reverse genetics system for 12-segmented double-stranded RNA viruses
Ryotaro Nouda, Shohei Minami, Yuta Kanai, Takahiro Kawagishi, Jeffery A. Nurdin, Moeko Yamasaki, Ryusei Kuwata, Hiroshi Shimoda, Ken Maeda, Takeshi Kobayashi
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 .