TY - JOUR
T1 - Entirely plasmid-based reverse genetics system for rotaviruses
AU - Kanai, Yuta
AU - Komoto, Satoshi
AU - Kawagishi, Takahiro
AU - Nouda, Ryotaro
AU - Nagasawa, Naoko
AU - Onishi, Misa
AU - Matsuura, Yoshiharu
AU - Taniguchi, Koki
AU - Kobayashi, Takeshi
N1 - Funding Information:
We thank Terence S. Dermody for reviewing the manuscript, Kaede Yukawa for secretarial work, Toru Okamoto for technical advice, Saori Fukuda for technical assistance, Polly Roy for providing BSR cells, and Naoto Ito for providing BHK/T7-9 cells. This work was supported in part by grants-in-aid for the Research Program on Emerging and Re-Emerging Infectious Diseases from the Japan Agency for Medical Research and Development and Japanese Society for the Promotion of Science (JSPS) KAKENHI Grants JP16K19138, JP15J04209, and JP26292149.
Publisher Copyright:
© 2017, National Academy of Sciences. All rights reserved.
PY - 2017/2/28
Y1 - 2017/2/28
N2 - 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 small transmembrane protein that accelerates cell-to-cell fusion and vaccinia virus capping enzyme. We used this system to obtain insights into the process by which RV nonstructural protein NSP1 subverts host innate immune responses. By insertion into the NSP1 gene segment, we recovered recombinant viruses that encode split-green fluorescent protein-tagged NSP1 and NanoLuc luciferase. This technology will provide opportunities for studying RV biology and foster development of RV vaccines and therapeutics.
AB - 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 small transmembrane protein that accelerates cell-to-cell fusion and vaccinia virus capping enzyme. We used this system to obtain insights into the process by which RV nonstructural protein NSP1 subverts host innate immune responses. By insertion into the NSP1 gene segment, we recovered recombinant viruses that encode split-green fluorescent protein-tagged NSP1 and NanoLuc luciferase. This technology will provide opportunities for studying RV biology and foster development of RV vaccines and therapeutics.
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U2 - 10.1073/pnas.1618424114
DO - 10.1073/pnas.1618424114
M3 - Article
C2 - 28137864
AN - SCOPUS:85014364320
VL - 114
SP - 2343
EP - 2348
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 9
ER -