PNAS USA, 2017 Jan 30.2017/01/30
Rotaviruses (RVs) are highly important pathogens that cause severe diarrhea among infants and young children worldwide. The understanding of the molecular mechanisms underlying RV replication and pathogenesis has been hampered by the lack of an entirely plasmid-based reverse genetics system. In this study, we describe the recovery of recombinant RVs entirely from cloned cDNAs. The strategy requires coexpression of a fusion-associated small transmembrane (FAST) protein that accelerates cell-to-cell fusion and vaccinia virus capping enzyme. We used this system to obtain new insights into the process by which RV NSP1 subverts host innate immune responses. By insertion into the NSP1 gene segment, we recovered recombinant viruses that encode split-GFP-tagged NSP1 and NanoLuc luciferase. This technology will provide new opportunities for studying RV biology and foster development of new RV vaccines and therapeutics.