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Engineering and analysis of mouse gene clusters by exploiting local hopping characteristic of transposon

Research Project

Project/Area Number 18500327
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory animal science
Research InstitutionOsaka University

Principal Investigator

KOKUBU Chikara  Osaka University, Graduate School of Medicine, Assistant Professor (70379238)

Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥4,110,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥510,000)
Fiscal Year 2007: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
KeywordsDevelopmental engineering / Chromosomal engineering / cis-regulatory element / Enhancer / Morphogenesis / Transposon / Mouse / Heterochromatin
Research Abstract

Large cisregulatory landscapes are a remarkable feature of vertebrate genomes, particularly seen at key developmental gene loci with finely tuned expression patterns. Existing genetic tools for surveying such a large genomic region of interest, ranging from a few kb to a few Mb, are limited. Here we propose a chromosomal engineering strategy exploiting the local hopping property of the Sleeping Beauty (SB) transposon in the mouse genome. We devised the transposon-based Local Hopping Enhancer Detector (LHED) vector carrying an enhancer-detection LacZ reporter and loxP cassette, which allows monitoring of transcription-enhancing activities in the native genomic context and Cre-mediated deletion/inversion rearrangement between the transposon donor and acceptor sites.
As a proof of principle, we generated embryonic stem (ES) cells with a targeted integration of the LHED vector into the developmentally and evolutionally critical Paxl gene locus, followed by efficient local transposition, nested deletion formation, and rapid derivation of embryos by tetraploid complementation. Whole-mount reporter expression analysis of the engineered embryos showed different expression patterns corresponding to each insertion/deletion configuration and markedly facilitated long-range cis-regulatory element mapping in this genomic neighborhood, demonstrating the great potential of this experimental platform for functional exploration of very focused areas in the mouse genome (manuscript in revision).

Report

(3 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • Research Products

    (12 results)

All 2008 2007

All Journal Article (3 results) (of which Peer Reviewed: 1 results) Presentation (9 results)

  • [Journal Article] Sleeping beauty transposase has an affinity for heterochromatin conformation2007

    • Author(s)
      Ikeda R., et al
    • Journal Title

      Mol Cell Biol. 27(5)

      Pages: 1665-1676

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Sleeping beauty transposase has an affinity for heterochromatin conformation.2007

    • Author(s)
      Ikeda R, Kokubu C, Yusa K, Keng VW, Horie K, Takeda J.
    • Journal Title

      Mol Cell Biol. 27

      Pages: 1665-1676

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Sleeping beauty transposase has an affinity for heterochromatin conformation2007

    • Author(s)
      Ikeda R. et al.
    • Journal Title

      Mol Cell Biol. 27(5)

      Pages: 1665-1676

    • Related Report
      2006 Annual Research Report
  • [Presentation] Mapping by Hopping : A Transposon-Based Chromosomal Engineering to Survey a Large Cis-Regulatory Landscape in the Mouse Genome2008

    • Author(s)
      Kokubu C.
    • Organizer
      The 16th CDB Meeting : Cis-Sequence Regulation and its Evolution
    • Place of Presentation
      RIKEN-CDB. Kobe, JAPAN.
    • Year and Date
      2008-09-30
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Mapping by Hopping: A Transposon-Based Chromosomal Engineering to Survey a Large Cis-Regulatory Landscape in the Mouse Genome.2008

    • Author(s)
      Kokubu C.
    • Organizer
      The 16th CDB Meeting: Cis-Sequence Regulation and its Evolution.
    • Place of Presentation
      Riken-CDB, Kobe, JAPAN.
    • Year and Date
      2008-09-30
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Transposon-Based Genome Engineering to Survey a Large Cis-Regulatory Landscape.2007

    • Author(s)
      Kokubu C.
    • Organizer
      Building a Better Mouse II Conference
    • Place of Presentation
      Vanderbilt University, Nashville, USA.
    • Year and Date
      2007-07-14
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Transposon-Based Genome Engineering to Survey a Large Cis-Regulatory Landscape.2007

    • Author(s)
      Kokubu C.
    • Organizer
      Building a Better Mouse II Conference.
    • Place of Presentation
      Vanderbilt University, Nashville, USA.
    • Year and Date
      2007-07-14
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] トランスポゾンを用いたマウス長距離シス調節ゲノム領域の改変/解析法の開発2007

    • Author(s)
      國府 力
    • Organizer
      第40回日本発生生物学会・第59回日本細胞生物学会合同大会
    • Place of Presentation
      福岡国際会議場
    • Year and Date
      2007-05-30
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
  • [Presentation] Mapping by Hopping: A Transposon-Based Chromosomal Engineering Strategy to Survey a Large Cis-Regulatory Landscape in the Mouse Genome.2007

    • Author(s)
      Kokubu C.
    • Organizer
      Joint Meeting of the Japanese Society of Developmental Biologists & the Japan Society for Cell Biology.
    • Place of Presentation
      Fukuoka, JAPAN.
    • Year and Date
      2007-05-30
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Sleeping Beautyトランスポゾンを用いた挿入変異体作製と染色体エンジニアリング2007

    • Author(s)
      國府 力
    • Organizer
      第54回日本実験動物学会総会
    • Place of Presentation
      タワーホール船堀(東京)
    • Year and Date
      2007-05-25
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
  • [Presentation] Insertional mutagenesis and chromosomal engineering using the Sleeping Beauty transposon2007

    • Author(s)
      Kokubu C.
    • Organizer
      The 54th Annual Meeting of the Japanese Association for Laboratory Animal Science.
    • Place of Presentation
      Tokyo, JAPAN.
    • Year and Date
      2007-05-25
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Transposon-Based Genome Engineering to Survey a Large Cis-Reaulatory Landscape2007

    • Author(s)
      Kokubu C.
    • Organizer
      Building a Better Mouse II Conference
    • Place of Presentation
      Vanderbilt University, Nashville, USA.
    • Related Report
      2007 Annual Research Report

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Published: 2006-04-01   Modified: 2016-04-21  

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