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Development of a rice gene targeting with autonomous self-marker free system as a new breeding technique

Research Project

Project/Area Number 16K07560
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Science in genetics and breeding
Research InstitutionMeijo University

Principal Investigator

Terada Rie  名城大学, 農学部, 教授 (30137799)

Research Collaborator SHIMATANI zenpei  
KAWANO yoji  
ARAKAWA masao  
Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Keywordsゲノム編集 / ターゲティング / イネ / トランスポゾン / OsRacGEF1 / 耐病性 / OsRac1GEF1 / 育種学 / 遺伝学 / 遺伝子 / ゲノム / 植物バイオテクノロジ- / バイオテクノロジ-
Outline of Final Research Achievements

We have developed a gene targeting (GT) system with an autonomous self-marker free as a new breeding technique. In this system the positive selection marker of homologous recombination (HR) mediated GT is placed in maize DNA transposon, Ac/Ds and by induction of Ac expression just after GT the positive selection marker is eliminated from targeted gene locus. Since pseud-phosphorylation of OsRacGEF1 was found increase the resistance to blast fungus, we modified OsRacGEF1-S549D by GT and subsequently the positive selection marker was eliminated by Ac activation. TG rice lines of OsRacGEF1-S549D homozygote without any artificial marker sequences were obtained, which revealed increased resistance to both of blast fungus and sheath blight disease.

Academic Significance and Societal Importance of the Research Achievements

イネ遺伝子ターゲティング法は標的遺伝子のみを自在に改変でき既に30以上の遺伝子改変に成功した。ターゲティング遺伝子改変とAc/Dsによる自律的選抜マーカー削除による迅速育種法では、ポジティブ選抜マーカーの削除によって複数遺伝子のターゲティング改変が可能で、分子遺伝子学の最新知見を活用した複数の遺伝子編集によるゲノムデザインが可能であり、圃場展開も視野に入れた信頼性の高い実用化イネを1年弱の短期間に作り出す迅速次世代育種法として利用促進が期待できる。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (12 results)

All 2019 2018 2017 Other

All Journal Article (3 results) (of which Peer Reviewed: 3 results,  Open Access: 2 results,  Acknowledgement Compliant: 1 results) Presentation (6 results) Remarks (1 results) Patent(Industrial Property Rights) (2 results)

  • [Journal Article] Rice Gene Targeting by Homologous Recombination with a Positive-Negative Selection Strategy2019

    • Author(s)
      Terada R and Shimatani Z
    • Journal Title

      Methods in Molecular Biology

      Volume: in press

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Inheritance of co-edited genes by CRISPR-based targeted nucleotide substitutions in rice2018

    • Author(s)
      Shimatani Zenpei、Fujikura Ushio、Ishii Hisaki、Matsui Yusuke、Suzuki Minoru、Ueke Yuki、Taoka Ken-ichiro、Terada Rie、Nishida Keiji、Kondo Akihiko
    • Journal Title

      Plant Physiology and Biochemistry

      Volume: 131 Pages: 78-83

    • DOI

      10.1016/j.plaphy.2018.04.028

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion2017

    • Author(s)
      Shimatani, Z., Kashojiya, S., Takayama, M., Terada, R., Arazoe, T., Ishii, H., Teramura, H., Yamamoto, T., Komatsu, H., Miura, K., Ezura, H., Nishida, K., Ariizumi, T. and Kondo A.
    • Journal Title

      Nature Biotechnology

      Volume: 印刷中 Issue: 5 Pages: 441-443

    • DOI

      10.1038/nbt.3833

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] イネbZIP型転写因子によるブラシノステロイド関連遺伝子発現制御の解析2019

    • Author(s)
      吉田英樹, 島谷善平,鈴木寿法, 寺田理枝,上口(田中)美弥子,松岡信,辻寛之
    • Organizer
      第60回日本植物生理学会年会(名古屋)
    • Related Report
      2018 Annual Research Report
  • [Presentation] イネ遺伝子ターゲティングに基づく人工配列を残さない標的配列の精密ゲノム編集2019

    • Author(s)
      松井祐介, 島谷善平, 寺田理枝
    • Organizer
      第60回日本植物生理学会年会(名古屋)
    • Related Report
      2018 Annual Research Report
  • [Presentation] 高感度な発光レポーターNanoLucの植物細胞での利用2019

    • Author(s)
      田岡健一郎,島谷善平,小川真奈,齋藤洋美,池田洋一,赤司裕子,山口公志,寺田理枝,川崎努,辻寛之
    • Organizer
      第60回日本植物生理学会年会(名古屋)
    • Related Report
      2018 Annual Research Report
  • [Presentation] イネ茎頂分裂組織の DNA メチル化パターンの動態と制御機構の解析2018

    • Author(s)
      肥後あすか,才原徳子,三浦史仁,東陽子,山田恵美,玉置祥二郎,伊藤佑,樽谷芳明,坂本智昭, 藤原正幸,倉田哲也,深尾陽一朗,森藤暁,寺田理枝,伊藤隆司,角谷徹仁,島本功,辻寛之
    • Organizer
      第59回日本植物生理学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] いもち病耐性イネ育種のための自律的マーカー削除可能な標的遺伝子編集2017

    • Author(s)
      鈴木 実、植家優紀、河野洋治、島本 功、荒川征夫、寺田理枝、島谷善平
    • Organizer
      日本育種学会第131回講演会
    • Place of Presentation
      名古屋大学
    • Related Report
      2016 Research-status Report
  • [Presentation] イネ花成誘導遺伝子のin-vivo発現解析を目指したノックイン・ノックアウトの三重改変2017

    • Author(s)
      植家優紀・鈴木 実・辻 寛之・田岡健一郎・島本 功・寺田理枝・島谷善平
    • Organizer
      日本育種学会第131回講演会
    • Place of Presentation
      名古屋大学
    • Related Report
      2016 Research-status Report
  • [Remarks] 名城大学 育て達人 第149回

    • URL

      http://www.meijo-u.ac.jp/sp/sodate/detail.html?id=RO6fwI

    • Related Report
      2016 Research-status Report
  • [Patent(Industrial Property Rights)] 植物細胞のゲノム編集用核酸及びその用途2019

    • Inventor(s)
      島谷善平, 寺田理枝
    • Industrial Property Rights Holder
      国立大学法人神戸大学,学校法人名城大学
    • Industrial Property Rights Type
      特許
    • Filing Date
      2019
    • Related Report
      2018 Annual Research Report
  • [Patent(Industrial Property Rights)] 植物体に用いる組換えベクター及びその利用2018

    • Inventor(s)
      寺田理枝, 島谷善平
    • Industrial Property Rights Holder
      学校法人名城大学
    • Industrial Property Rights Type
      特許
    • Filing Date
      2018
    • Acquisition Date
      2018
    • Related Report
      2018 Annual Research Report

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Published: 2016-04-21   Modified: 2023-01-30  

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