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Establishment of a strategy for generation of Orthobunyavirus expressing an enhanced foreign reporter gene

Research Project

Project/Area Number 17K08074
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Veterinary medical science
Research InstitutionThe University of Tokyo

Principal Investigator

Uema Akiko  東京大学, 大学院農学生命科学研究科(農学部), 特任助教 (20630156)

Co-Investigator(Kenkyū-buntansha) 堀本 泰介  東京大学, 大学院農学生命科学研究科(農学部), 教授 (00222282)
村上 晋  東京大学, 大学院農学生命科学研究科(農学部), 准教授 (10636757)
Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2017: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Keywords改変型組換えアカバネウイルス / 転写プロモーター活性 / 感染の可視化 / 改変型オルソブニヤウイルス / 改変型AKAV / 転写プロモーター活性の増強 / 病原微生物
Outline of Final Research Achievements

Although the Akabane virus (AKAV) expressing enhanced green fluorescence protein, we previously generated, was able to detect infected cells in in vivo histopathological study, its fluorescent signal was too weak to apply to in vivo imaging study. Here, we successfully generated a modified reporter virus. The produced virus expressed higher intensity of eGFP fluorescence both in vitro and in vivo than the original virus did. In addition, the virus was pathogenic in mice at a comparable level to that in the original and wild-type virus. In the mice infected with the virus, the fluorescent signals, i.e., the virus-infected cells, were detected in the central nervous system using the whole-organ imaging. Our findings indicate that the modified virus could be used as a powerful tool to help elucidate the dynamics of AKAV in vivo.

Academic Significance and Societal Importance of the Research Achievements

本研究で行なった蛍光発現アカバネウイルスの改変では、アンチゲノムからの転写やゲノムへの複製が増強されていることが分かった。これまでに報告されたオルソブニヤウイルスのミニゲノムアッセイを使った研究とは逆の結果となっており、オルソブニヤウイルスゲノムに関する新たな知見が得られた。
本研究で確立されたウイルス作出手法により、蛍光遺伝子を別の病原体の遺伝子に置き換えることで、アカバネウイルスの多価ワクチン製造が可能となる。またこの手法は別のウイルスにも応用が期待でき、感染機構の解明に貢献できる。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • Research Products

    (3 results)

All 2019 2018 Other

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

  • [Journal Article] Generation of a GFP reporter Akabane virus with enhanced fluorescence intensity by modification of artificial ambisense S genome.2019

    • Author(s)
      Takenaka-Uema A, Murakami S, Ushio N, Kobayashi-Kitamura T, Uema M, Uchida K, Horimoto T.
    • Journal Title

      Viruses

      Volume: 11(7) Issue: 7 Pages: 634-634

    • DOI

      10.3390/v11070634

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] 蛍光高発現組換えアカバネウイルスの性状解析2018

    • Author(s)
      上間亜希子
    • Organizer
      第66回日本ウイルス学会学術集会
    • Related Report
      2018 Research-status Report
  • [Remarks] 緑色蛍光を高度に発現する人工アンビセンス鎖ゲノムをもつ組換えアカバネウイルスの作出

    • URL

      https://www.a.u-tokyo.ac.jp/topics/topics_20190718-1.html

    • Related Report
      2019 Annual Research Report

URL: 

Published: 2017-04-28   Modified: 2021-02-19  

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