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Generation of synthetic rescue system in murine haploid ES cells.

Research Project

Project/Area Number 26640124
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Medical genome science
Research InstitutionOsaka University

Principal Investigator

TAKEDA Junji  大阪大学, 医学(系)研究科(研究院), 教授 (50163407)

Project Period (FY) 2014-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2014: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Keywordsゲノムワイド関連解析 / ハプロイドES細胞
Outline of Final Research Achievements

We performed experiments to generate a novel synthetic rescue system in murine haploid ES cells. For conditional KO system in haploid ES cells, we first tried to insert tamoxifen- inducible cassette, ERT2-iCre-ERT2 into the Rosa 26 locus. Since homologous targeting in haploid ES cells was very low, we used ZFN for the targeting.
Meanwhile, production of CRISPR-gRNA library has bee reported. The library is able to disrupt both alleles of a gene of interest.
We modified both alleles of Oct4 to floxed ones and we introduced CRISPR-gRNA library to disrupt endogenous genes randomly. After disruption of Oct4 by activation of Cre with tamoxifen. We isolated undifferentiated clones. We are analyzing these clones right now.

Report

(2 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • Research Products

    (4 results)

All 2014

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (3 results) (of which Invited: 2 results)

  • [Journal Article] Simulation and estimation of gene number in a biological pathway using almost complete saturation mutagenesis screening of haploid mouse cells2014

    • Author(s)
      Tokunaga M, Kokubu C, Maeda Y, Sese J, Horie K, Sugimoto N, Kinoshita T, Yusa K, Takeda J.
    • Journal Title

      BMC Genomics

      Volume: 15 Issue: 1 Pages: 1016-1016

    • DOI

      10.1186/1471-2164-15-1016

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] Generation of gene-trapped haploid ES cell bank2014

    • Author(s)
      竹田 潤二
    • Organizer
      Systems Bioloby of Gene Regulation and Genome Editing
    • Place of Presentation
      蘇州(中国)
    • Year and Date
      2014-09-09 – 2014-09-12
    • Related Report
      2014 Annual Research Report
  • [Presentation] 哺乳動物において順遺伝学的手法は有用か?2014

    • Author(s)
      竹田 潤二
    • Organizer
      第33回分子病理学研究会
    • Place of Presentation
      宮城県(日本)
    • Year and Date
      2014-07-26
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] ハプロイドES細胞を利用した遺伝子探索2014

    • Author(s)
      石田 靖雅
    • Organizer
      第66回日本細胞生物学会大会
    • Place of Presentation
      奈良県(日本)
    • Year and Date
      2014-06-11
    • Related Report
      2014 Annual Research Report
    • Invited

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Published: 2014-04-04   Modified: 2016-06-03  

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