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Development of screening system for bone forming reagents by using TALEN genome engineering technique

Research Project

Project/Area Number 26670807
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Functional basic dentistry
Research InstitutionOsaka University

Principal Investigator

Nishimura Riko  大阪大学, 歯学研究科(研究院), 教授 (60294112)

Co-Investigator(Renkei-kenkyūsha) HATA KENJI  大阪大学, 大学院歯学研究科, 准教授 (80444496)
MURAKAMI TOMOHIKO  大阪大学, 大学院歯学研究科, 講師 (50510723)
Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywords骨形成 / ゲノム編集 / ゲノム編集法
Outline of Final Research Achievements

In order to develop effective screening system for bone forming reagents, we attempted to generate cell lines knock-ined both Venus and luciferase genes by using the Golden Gate TALEN genome engineering technique. Although the Golden Gate TALEN genome engineering technique is much more effective and useful than conventional TALEN system, selection of positive clone cells was still complicate. We, therefore, attempted to optimize a new genome engineering technique, PITCh method, which would be very powerful for knock-in of interested genes into to mammalian cells. First, we used Cas9-based PITCh technique, allowing us to generate guide RNA construct easily. Second, we designed and generated a PITCh donor vector, in which we introduced CLuc luciferase, Puromycin resistant genes with EGFP expression cassette. This optimization seems very effective and helpful for selection of the positive clone cells.

Report

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

    (3 results)

All 2015 2014

All Presentation (2 results) (of which Invited: 1 results) Book (1 results)

  • [Presentation] ゲノム編集技術を用いた新規プロテインキナーゼの軟骨細胞分化における役割解明2015

    • Author(s)
      森 裕美子、萩野 弘将、後藤 満帆、長田 奈幹、高畑 佳史、村上 智彦、波多 賢二、西村 理行
    • Organizer
      第33回日本骨代謝学会
    • Place of Presentation
      京王プラザホテル東京(東京都新宿区)
    • Year and Date
      2015-07-23
    • Related Report
      2015 Annual Research Report
  • [Presentation] 骨格形成過程における転写制御システムの役割2014

    • Author(s)
      西村 理行
    • Organizer
      歯科基礎医学会
    • Place of Presentation
      福岡国際会議場
    • Year and Date
      2014-09-26
    • Related Report
      2014 Research-status Report
    • Invited
  • [Book] 骨ペディア(分担執筆)2015

    • Author(s)
      西村 理行
    • Total Pages
      327
    • Publisher
      羊土社
    • Related Report
      2015 Annual Research Report

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Published: 2014-04-04   Modified: 2017-05-10  

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