Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 2343-2348 |
| Number of pages | 6 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 114 |
| Issue number | 9 |
| DOIs | |
| Publication status | Published - 28-02-2017 |
Fingerprint
All Science Journal Classification (ASJC) codes
- General
Cite this
}
Entirely plasmid-based reverse genetics system for rotaviruses. / Kanai, Yuta; Komoto, Satoshi; Kawagishi, Takahiro; Nouda, Ryotaro; Nagasawa, Naoko; Onishi, Misa; Matsuura, Yoshiharu; Taniguchi, Koki; Kobayashi, Takeshi.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, No. 9, 28.02.2017, p. 2343-2348.Research output: Contribution to journal › Article
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
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.
UR - http://www.scopus.com/inward/record.url?scp=85014364320&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85014364320&partnerID=8YFLogxK
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 -