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Establishment of highly efficient chloroplast genome editing by using synthetic custom design nucleases

Research Project

Project/Area Number 25450002
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Science in genetics and breeding
Research InstitutionThe University of Tokushima

Principal Investigator

Osakabe Keishi  徳島大学, 農工商連携センター, 特任教授 (70450335)

Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2013: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Keywords葉緑体 / ゲノム編集 / TALEN / Cas9 / タバコ / CRISPR / CRISPR/CAS9 / 遺伝子ターゲッティング
Outline of Final Research Achievements

To establish chloroplast genome editing in order to open a frontier of chloroplast engineering, site-specific DNA cleavage system by using the synthetic custom design nuclease was designed and constructed. At first, TALEN as a synthetic custom design nuclease was chosen, and The TALEN with transit peptide (tp-TALEN) against to the target on chloroplast genome was constructed. The tp-TALEN could cut the target DNA specifically in vitro, but was failed to transport to chloroplast. Therefore, Cas9 nuclease was used instead of TALEN. Cas9 with transit peptide was successively transported to chloroplast.

Report

(4 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • 2013 Research-status Report
  • Research Products

    (12 results)

All 2016 2015 2014 2013

All Journal Article (5 results) (of which Peer Reviewed: 4 results,  Acknowledgement Compliant: 3 results,  Open Access: 2 results) Presentation (6 results) Book (1 results)

  • [Journal Article] Optimization of CRISPR/Cas9 genome editing to modify abiotic stress responses in plants.2016

    • Author(s)
      Yuriko Osakabe, Takahito Watanabe, Shigeo S Sugano, Risa Ueta, Ryosuke Ishihara, Kazuo Shinozaki, Keishi Osakabe
    • Journal Title

      Scientific Reports

      Volume: -

    • NAID

      120006240041

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Genome engineering of woody plants: past, present and future2016

    • Author(s)
      Yuriko Osakabe, Shigeo S Sugano and Keishi Osakabe
    • Journal Title

      Journal of Wood Science

      Volume: 62 Issue: 3 Pages: 1-9

    • DOI

      10.1007/s10086-016-1548-5

    • NAID

      120006961148

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Genome editing with engineered nucleases in plants.2015

    • Author(s)
      Yuriko Osakabe and Keishi Osakabe
    • Journal Title

      Plant & Cell Physiology

      Volume: 56 Issue: 3 Pages: 389-400

    • DOI

      10.1093/pcp/pcu170

    • Related Report
      2015 Annual Research Report 2014 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] A mutated cytosine deaminase gene, codA (D314A), as an efficient negative selection marker for gene targeting in rice.2014

    • Author(s)
      Keishi Osakabe, Ayako Nishizawa-Yokoi, Namie Ohtsuki, Yuriko Osakabe, Seiichi Toki
    • Journal Title

      Plant Cell Physiology

      Volume: 55 Issue: 3 Pages: 658-665

    • DOI

      10.1093/pcp/pct183

    • NAID

      40020302254

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] 人工エンドヌクレアーゼを利用した高等植物ゲノム改変技術の新展開2013

    • Author(s)
      刑部敬史、刑部祐里子
    • Journal Title

      細胞工学

      Volume: 32 Pages: 520-525

    • Related Report
      2013 Research-status Report
  • [Presentation] ゼニゴケにおける高効率ゲノム編集ベクターの開発2016

    • Author(s)
      菅野茂夫,西浜竜一,白川一,松田頼子,髙木純平,西村いくこ,刑部敬史,河内孝之
    • Organizer
      第57回日本植物生理学会大会
    • Place of Presentation
      岩手大学上田キャンパス(岩手県盛岡市)
    • Year and Date
      2016-03-16
    • Related Report
      2015 Annual Research Report
  • [Presentation] CRISPR/Cas9によるシロイヌナズ ナ環境ストレス応答性遺伝子のゲ ノム編集2016

    • Author(s)
      刑部 祐里子, 菅野 茂夫, 渡辺 崇人, 上田 梨紗, 石原 諒典, 篠崎 一雄, 刑部 敬史
    • Organizer
      第57回日本植物生理学会大会
    • Place of Presentation
      岩手大学上田キャンパス(岩手県盛岡市)
    • Year and Date
      2016-03-16
    • Related Report
      2015 Annual Research Report
  • [Presentation] 高効率 CRISPR/Cas9による SlIAA9 ノックアウトトマトの作出2016

    • Author(s)
      上田 梨紗, 阿部 千尋, 石原 諒典, 渡辺 崇人, 菅野 茂夫, 宮脇 克行, 野地 澄晴, 刑部 祐里子, 刑部 敬史
    • Organizer
      第57回日本植物生理学会大会
    • Place of Presentation
      岩手大学上田キャンパス(岩手県盛岡市)
    • Year and Date
      2016-03-16
    • Related Report
      2015 Annual Research Report
  • [Presentation] CRISPR/Cas9によるトマトIAA9 遺伝子を標的としたゲノム編集技術の確立2015

    • Author(s)
      上田 梨紗, 石原 諒典, 阿部 千尋, 渡辺 崇人, 菅野 茂夫, 宮脇 克行, 野地 澄晴, 刑部 祐里子, 刑部 敬史
    • Organizer
      第38回日本分子生物学会年会
    • Place of Presentation
      神戸ポートアイランド(兵庫県神戸市)
    • Year and Date
      2015-12-01
    • Related Report
      2015 Annual Research Report
  • [Presentation] CRISPR/Cas9によるトマトIAA9遺伝子を標的としたゲノム編集2015

    • Author(s)
      上田梨紗、石原諒典、渡辺崇人、菅野茂夫、宮脇克行、野地澄晴、刑部祐里子、刑部敬史
    • Organizer
      第56回日本植物生理学会年会
    • Place of Presentation
      東京農業大学( 東京都世田谷区)
    • Year and Date
      2015-03-16 – 2015-03-18
    • Related Report
      2014 Research-status Report
  • [Presentation] CRISPR/Cas9による植物ゲノム編集技術開発と環境応答能の改変2015

    • Author(s)
      刑部祐里子、菅野茂夫、篠崎一雄、野地澄晴、刑部敬史
    • Organizer
      第56回日本植物生理学会年会
    • Place of Presentation
      東京農業大学( 東京都世田谷区)
    • Year and Date
      2015-03-16 – 2015-03-18
    • Related Report
      2014 Research-status Report
  • [Book] Targeted Genome Editing Using Engineered Nucleases: ZFNs, TALENs, and the CRISPR/Cas9 System2015

    • Author(s)
      Yuriko Osakabe and Keishi Osakabe
    • Total Pages
      14
    • Publisher
      Springer -Verlag
    • Related Report
      2014 Research-status Report

URL: 

Published: 2014-07-25   Modified: 2019-07-29  

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