Establishment of a Reverse Genetics System for Rotavirus: A Multisegmented, Double-Stranded RNA Virus

Abstract

Evaluation of: Komoto S, Sasaki J, Taniguchi K: Reverse genetics system for introduction of site-specific mutations into the double-stranded RNA genome of infectious rotavirus. Proc. Natl Acad. Sci. USA 103, 4646–4651 (2006). Rotavirus (RV), a double-stranded RNA virus within the Reoviridae family, is one of the most common causes of severe diarrhea in infants and young children worldwide. RV infections are highly contagious and may lead to severe dehydration and even death. Although vaccinations represent the most promising method for preventing RV disease in children, molecular virological techniques, such as reverse genetics, which provide a powerful tool not only for generating live-attenuated vaccines or vaccine vectors, but also for understanding gene functions and pathogenesis, are limited in the research field of RV. Komoto and colleagues have developed a novel reverse genetics system for RV using a traditional T7 RNA polymerase expression system. By using a strong selection method with neutralizing monoclonal antibodies specific for the helper RV strain KU VP4 protein, a KU-based transfectant RV carrying the strain SA11 VP4 segment, which is derived from the T7 RNA polymerase-driven plasmid, was recovered. The establishment of this system will provide a greater understanding of the molecular biology of RVs, including their replication and pathogenesis, as well as a tool for the development of attenuated vaccines or vaccine vectors.