A plasmid-based reverse genetics system for rotaviruses

Abstract

Rotavirus (RV), a non-enveloped icosahedral virus containing eleven gene segments of double-stranded RNA, is the leading cause of severe, acute diarrhea among infants and young children worldwide. Safe and effective rotavirus vaccines have been available since 2006, and have markedly reduced the number of deaths by severe gastroenteritis. However, rotaviruses are still responsible for approximately 200,000 deaths annually worldwide. Reverse genetics systems for the manipulation of viral genomes are a powerful approach for studying viral replication and pathogenesis, and for developing vaccines and viral vectors. The understanding of the molecular mechanisms underlying RV pathogenesis, or development of next generation vaccines, has been hampered by the lack of a complete reverse genetics system. Recently, we developed a novel reverse genetics system which enabled recovery of recombinant RVs entirely from cloned cDNAs. This new strategy requires co-expression of a small transmembrane protein that accelerates cell-to-cell fusion and vaccinia virus capping enzyme. In this review, the strategies and history of the development of reverse genetics systems for the family Reoviridae are described.