1887

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Volume 103, Issue 12

A reverse genetics system for human rotavirus G2P[4] No Access

Abstract

Rotaviruses (RVs) are an important cause of acute gastroenteritis in young children. Recently, versatile plasmid-based reverse genetics systems were developed for several human RV genotypes; however, these systems have not been developed for all commonly circulating human RV genotypes. In this study, we established a reverse genetics system for G2P[4] human RV strain HN126. Nucleotide sequence analysis, including that of the terminal ends of the viral double-stranded RNA genome, revealed that HN126 possessed a DS-1-like genotype constellation. Eleven plasmids, each encoding 11 gene segments of the RV genome, and expression plasmids encoding vaccinia virus RNA capping enzyme (D1R and D12L), Nelson Bay orthoreovirus FAST, and NSP2 and NSP5 of HN126, were transfected into BHK-T7 cells, and recombinant strain HN126 was generated. Using HN126 or simian RV strain SA11 as backbone viruses, reassortant RVs carrying the outer and intermediate capsid proteins (VP4, VP7 and VP6) of HN126 and/or SA11 (in various combinations) were generated. Viral replication analysis of the single, double and triple reassortant viruses suggested that homologous combination of the VP4 and VP7 proteins contributed to efficient virus infectivity and interaction between other viral or cellular proteins. Further studies of reassortant viruses between simian and other human RV strains will contribute to developing an appropriate model for human RV research, as well as suitable backbone viruses for generation of recombinant vaccine candidates.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): reverse genetics system and rotavirus
Funding
This study was supported by the:
  • Japan Science and Technology Agency Moonshot R&D (Award JPMJMS2025)
    • Principle Award Recipient: TakeshiKobayashi
  • KAKENHI (Award JP21H02739)
    • Principle Award Recipient: TakeshiKobayashi
  • AMED (Award JP20fk0108122)
    • Principle Award Recipient: TakeshiKobayashi
© 2022 The Authors
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2022-12-15
2024-04-19
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A reverse genetics system for human rotavirus G2P[4]
J. Gen. Virol. 103, 001816 (2022); https://doi.org/10.1099/jgv.0.001816
/content/journal/jgv/10.1099/jgv.0.001816
/content/journal/jgv/10.1099/jgv.0.001816
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