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Investigation of super-active TALEN in vitro

Research Project

Project/Area Number 25871134
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Molecular biology
Laboratory animal science
Research InstitutionThe Institute of Physical and Chemical Research

Principal Investigator

ITO Yoko  独立行政法人理化学研究所, 生命システム研究センター, 特別研究員 (60584571)

Project Period (FY) 2013-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
KeywordsTALEN / ゲノム編集 / 人工ヌクレアーゼTALEN
Outline of Final Research Achievements

Gene editing in vivo has become possible by the development of artificial nucleases that can be designed to cut the genome DNA selectively at the target site. TALENs are highly specific artificial nucleases. However in mammalian cells and embryos, TALENs often show poor activity. Recently we have developed “super-active” TALEN and demonstrated that which can mediate efficient genome editing in mouse embryos.
To evaluate how our “super-active” mutations actually affect enzymatic properties, we have produced active TALEN proteins and measured their activities in vitro. Our data suggested that super-active TALEN has high-affinity DNA-binding, which might cause high activity. In addition, structural biological approaches indicated the structural differences between super-active and conventional TALE. Our in vitro based approaches provide us a new insight for further improvement of TALEN techniques.

Report

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

    (4 results)

All 2015 2014 2013

All Presentation (4 results)

  • [Presentation] In vitro analysis of thermo stabilized super-active TALEN2015

    • Author(s)
      寺原(伊藤)陽子、池田一穂、宮下尚之、岡田康志
    • Organizer
      第120回 日本解剖学会・第92回日本生理学会 合同大会
    • Place of Presentation
      神戸国際会議場
    • Year and Date
      2015-03-21
    • Related Report
      2014 Annual Research Report
  • [Presentation] DNA結合ドメインの改変による高活性TALENの開発2014

    • Author(s)
      池田一穂、寺原陽子、岡田康志
    • Organizer
      第119回日本解剖学会
    • Place of Presentation
      栃木県下野市
    • Related Report
      2013 Research-status Report
  • [Presentation] TALE蛋白DNA結合ドメインの改変による高活性TALEN2013

    • Author(s)
      池田一穂、寺原陽子、岡田康志
    • Organizer
      第3回ゲノム編集研究会
    • Place of Presentation
      広島県東広島市
    • Related Report
      2013 Research-status Report
  • [Presentation] Optimization of TALE DNA binding domain for a high-efficiency genome editing in vivo2013

    • Author(s)
      Kazuho Ikeda, Yoko Terahara, Yasushi Okada
    • Organizer
      The American Society for Cell Biology Annual Meeting
    • Place of Presentation
      New Orleans America
    • Related Report
      2013 Research-status Report

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

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