Rotavirus is a member of the family Reoviridae, which have genomes consisting of 10-12 double-stranded RNA segments. The functions of proteins encoded by each segment of the rotavirus genome have been studied extensively by several methods including reassortants, temperature-sensitive mutants, isolates with rearranged RNA segments, RNAi analysis, and other procedures. However, as found for most RNA viruses, the technique of reverse genetics is required for precise genotype/phenotype correlation, for the analysis of the role of specific mutation in replication process and pathogenesis, and for the development of vectors and vaccines. In 2006, we presented the first description of a reverse genetics system for rotavirus, although a helper virus and a selection system are required. Since then, two other approaches have been reported for rotavirus reverse genetics, both requiring the presence of a helper virus. A tractable, helper virus-free reverse genetics system for rotavirus has not been developed so far, in contrast to the recent developments of plasmid only-based reverse genetics systems for other members of the Reoviridae.
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