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Screening of factors affecting gene targeting efficiency.

Research Project

Project/Area Number 24780010
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Breeding science
Research InstitutionNational Institute of Agrobiological Sciences

Principal Investigator

ENDO Masaki  独立行政法人農業生物資源研究所, ゲノム機能改変研究ユニット, 研究員 (40546371)

Project Period (FY) 2012-04-01 – 2014-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2013: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2012: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywords形質転換 / ジーンターゲッティング / 植物 / アグロバクテリウム / DNA二重鎖切断
Research Abstract

Arabidopsis tester lines possessing an I-SceI recognition site and an inducible I-SceI expression system were established for monitoring protein attachment at DNA double strand break (DSB) site and for monitoring gene targeting events. By using this experimental material, it was revealed that gene targeting efficiency was increased by a factor of 5-10 by DSB induction during transformation of gene targeting vector.
It is known that T-DNAs preferentially integrate at DNA DSB sites. Previous studies show that VirD2, which is one of the key Agrobacterium tumefaciens proteins involved in T-DNA processing and transfer is important for efficient T-DNA integration in to the plant genome. However there is no direct evidence to show the role of VirD2 in T-DNA integration step. For detecting in vivo VirD2 attachment at DSB sites, I established chromatin immunoprecipitation assay using protoplast of this Arabidopsis tester line with transfection of VirD2 expression cassette and DSB induction.

Report

(3 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • Research Products

    (9 results)

All 2014 2013 2012 Other

All Journal Article (3 results) Presentation (6 results)

  • [Journal Article] 植物におけるゲノム編集2014

    • Author(s)
      遠藤真咲、土岐精一
    • Journal Title

      遺伝

      Volume: 68(2) Pages: 135-139

    • Related Report
      2013 Final Research Report
  • [Journal Article] 植物ゲノムの標的遺伝子改変における人工制限酵素の利用2013

    • Author(s)
      遠藤真咲、土岐精一
    • Journal Title

      植物工学

      Volume: 91 Pages: 448-451

    • Related Report
      2013 Final Research Report
  • [Journal Article] 植物ゲノムの標的遺伝子改変における人工制限酵素の利用2013

    • Author(s)
      遠藤真咲、土岐精一
    • Journal Title

      生物工学

      Volume: 91 Pages: 448-448

    • Related Report
      2013 Annual Research Report
  • [Presentation] 組換えの鋳型をゲノムから切り出す標的組換えシステム2013

    • Author(s)
      遠藤真咲、三上雅史、土岐精一
    • Organizer
      日本植物細胞分子生物学会第31回講演会
    • Place of Presentation
      北海道大学
    • Year and Date
      2013-09-10
    • Related Report
      2013 Final Research Report
  • [Presentation] 鋳型をゲノムから切り出すジーンターゲッティング法2013

    • Author(s)
      遠藤真咲、Stanton B Gelvin,土岐精一
    • Organizer
      日本育種学会第123回講演会
    • Place of Presentation
      東京農業大学
    • Year and Date
      2013-03-27
    • Related Report
      2013 Final Research Report
  • [Presentation] DNA double strand breaks and targeted T-DNA integration in Arabidopsis2012

    • Author(s)
      Endo M, Gelvin S, Toki S.
    • Organizer
      第35回日本分子生物学会年会
    • Place of Presentation
      福岡国際会議場・マリンメッセ福岡
    • Year and Date
      2012-12-13
    • Related Report
      2013 Final Research Report
  • [Presentation] 組換えの鋳型をゲノムから切り出す標的組換えシステム

    • Author(s)
      遠藤真咲、三上雅史、土岐精一
    • Organizer
      日本植物細胞分子生物学会 第31回講演会
    • Place of Presentation
      北海道大学
    • Related Report
      2013 Annual Research Report
  • [Presentation] 鋳型をゲノムから切り出すジーンターゲッティング法

    • Author(s)
      遠藤 真咲, Stanton B. Gelvin, 土岐 精一
    • Organizer
      日本育種学会 第123回講演会
    • Place of Presentation
      東京農業大学
    • Related Report
      2012 Research-status Report
  • [Presentation] DNA double strand breaks and targeted T-DNA integration in Arabidopsis

    • Author(s)
      Endo M, Gelvin S, Toki S
    • Organizer
      第35回日本分子生物学会年会
    • Place of Presentation
      福岡国際会議場・マリンメッセ福岡
    • Related Report
      2012 Research-status Report

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Published: 2013-05-31   Modified: 2019-07-29  

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