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Applications of entirely plasmid-based reverse genetics system on rotavirus research

Research Project

Project/Area Number 18K07150
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 49060:Virology-related
Research InstitutionFujita Health University

Principal Investigator

Komoto Satoshi  藤田医科大学, 医学部, 准教授 (60367711)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywordsロタウイルス / リバースジェネティクス系 / 遺伝子操作系 / 11-プラスミドシステム / ヒトロタウイルス / リバースジェネティクス / 腸管指向性ウイルスベクター / リアソータント / 動物ロタウイルス / 組換えロタウイルス / 増殖能 / 病原性
Outline of Final Research Achievements

Reverse genetics technology allows one to engineer infectious viruses from cloned cDNAs at will. We developed an efficient reverse genetics system that makes it possible to generate infectious rotaviruses (RVAs) from just 11 T7-driven plasmids encoding an RVA genome when the quantity ration of the 2 rescue T7-driven plasmids for the NSP2 and NSP5 genes is increased by 3-fold in relation to that of the other 9 plasmids (11 plasmid-only system). Furthermore, it is now possible to rescue recombinant RVAs even with severely less efficient infectivity such as RVA vectors by using the 11 plasmid-only system, which has not been possible with the original RVA reverse genetics system. Finally, the successful generation of the first infectious human RVA was achieved when we made minor modification to the 11 plasmid-only system by increasing the trypsin concentration in the maintenance medium of the cell culture and applying the roller-culture technique to grow human RVAs.

Academic Significance and Societal Importance of the Research Achievements

11-プラスミドシステムの確立により、今日では多くの研究者が容易にRVを人工合成できる状況に至った。現在、世界中で11-プラスミドシステムと11-プラスミドシステムをベースとした遺伝子操作系を駆使して、ロタウイルスの増殖、病原性発現の機構解明といった基礎研究、そして安全性に優れた次世代ワクチンや腸管指向性ウイルスベクター開発といった臨床応用を目指した研究が活発に行われている。また、11-プラスミドシステムは、まだ生体における安全性が確認されていない因子を一切用いないため、臨床応用を進めるうえで明確なアドバンテージがある。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (9 results)

All 2021 2020 2019 2018

All Journal Article (6 results) (of which Peer Reviewed: 6 results,  Open Access: 1 results) Presentation (3 results) (of which Invited: 1 results)

  • [Journal Article] Strategy for generation of replication?competent recombinant rotaviruses expressing multiple foreign genes2021

    • Author(s)
      Hatazawa Riona、Fukuda Saori、Kumamoto Kanako、Matsushita Fumio、Nagao Shizuko、Murata Takayuki、Taniguchi Koki、Matsui Taei、Komoto Satoshi
    • Journal Title

      Journal of General Virology

      Volume: 102 Issue: 4 Pages: 1-9

    • DOI

      10.1099/jgv.0.001587

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Rapid generation of rotavirus single-gene reassortants by means of eleven plasmid-only based reverse genetics2020

    • Author(s)
      Fukuda S, Hatazawa R, Kawamura Y, Yoshikawa T, Murata T, Taniguchi K, Komoto S.
    • Journal Title

      J Gen Virol

      Volume: 101 Issue: 8 Pages: 806-815

    • DOI

      10.1099/jgv.0.001443

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Generation of recombinant rotaviruses from just 11 cDNAs encoding a viral genome2020

    • Author(s)
      Komoto S, Fukuda S, Hatazawa R, Murata T, Taniguchi K.
    • Journal Title

      Virus Res

      Volume: 286 Pages: 198075-198075

    • DOI

      10.1016/j.virusres.2020.198075

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Reverse genetics system for human rotaviruses2020

    • Author(s)
      Komoto S, Fukuda S, Murata T, Taniguchi K.
    • Journal Title

      Microbiol Immunol

      Volume: - Issue: 6 Pages: 401-406

    • DOI

      10.1111/1348-0421.12795

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Generation of infectious recombinant human rotaviruses from just 11 cloned cDNAs encoding the rotavirus genome2019

    • Author(s)
      Komoto S, Fukuda S, Kugita M, Hatazawa R, Koyama C, Katayama K, Murata T, Taniguchi K.
    • Journal Title

      J Virol.

      Volume: 93(8) Issue: 8

    • DOI

      10.1128/jvi.02207-18

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Generation of recombinant rotaviruses expressing fluorescent proteins using an optimized reverse genetics system2018

    • Author(s)
      Komoto S, Fukuda S, Ide T, Ito N, Sugiyama M, Yoshikawa T, Murata T, Taniguchi K.
    • Journal Title

      J Virol

      Volume: Epub ahead of print Issue: 13

    • DOI

      10.1128/jvi.00588-18

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Presentation] Eleven-plasmid-based reverse genetics platform for efficient generation of rotavirus reassortants2019

    • Author(s)
      Fukuda S, Hatazawa R, Yoshikawa T, Murata T, Taniguchi K, Komoto S.
    • Organizer
      第67回日本ウイルス学会学術集会
    • Related Report
      2019 Research-status Report
  • [Presentation] ロタウイルスの遺伝子操作系2019

    • Author(s)
      河本聡志
    • Organizer
      第60回日本臨床ウイルス学会学術集会
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] Development of efficient and simple reverse genetics system based on only rotavirus cDNAs2018

    • Author(s)
      Fukuda S, Ide T, Ito N, Sugiyama M, Yoshikawa T, Murata T, Taniguchi K, Komoto S.
    • Organizer
      第66回日本ウイルス学会学術集会
    • Related Report
      2018 Research-status Report

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Published: 2018-04-23   Modified: 2022-01-27  

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