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A multiple gene expression switch using a multi-integrase system on a mammalian artificial chromosome vector.

Research Project

Project/Area Number 25540134
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Life / Health / Medical informatics
Research InstitutionTottori University

Principal Investigator

Ohbayashi Tetsuya  鳥取大学, 生命機能研究支援センター, 准教授 (80348804)

Co-Investigator(Renkei-kenkyūsha) Nakamura Kazuomi  鳥取大学, 生命機能研究支援センター, プロジェクト研究員 (90598137)
Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywords人工染色体ベクター / インテグレース / レポーターシステム / ゲノム編集 / ゲノム改変 / ゲノムプログラム / 遺伝子発現
Outline of Final Research Achievements

Site-specific recombinases (SSR), such as Cre or Flp recombination target cassettes, have been successfully excised or inverted by a single SSR to regulate transgene expression. This study investigated the potential for expanding the multiple regulation of transgenes using three different integrase systems. We designed three excision cassettes that expressed luciferase, where the luciferase expression could be exchanged to a fluorescent protein by site-specific recombination. Each cassette that was regulated independently by a different integrase was connected in tandem and inserted into a mouse artificial chromosome (MAC) vector in CHO cells. By transient expression of integrase, the targeted luciferase activity was lost and fluorescence was activated. These results suggest that the combined use of these integrase systems in a defined locus on a MAC vector permits the multiple regulation of transgene expression and might contribute to genomic or cell engineering.

Report

(4 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • 2013 Research-status Report
  • Research Products

    (16 results)

All 2015 2014 2013 Other

All Journal Article (7 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 7 results,  Open Access: 5 results,  Acknowledgement Compliant: 1 results) Presentation (8 results) (of which Invited: 3 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Use of a Human Artificial Chromosome for Delivering Trophic Factors in a Rodent Model of Amyotrophic Lateral Sclerosis.2015

    • Author(s)
      Watanabe Y, Kazuki Y, Kazuki K, Ebiki M, Nakanishi M, Nakamura K, Yoshida Yamakawa M, Hosokawa H, Ohbayashi T, Oshimura M, Nakashima K.
    • Journal Title

      Mol Ther Nucleic Acids.

      Volume: 4 Pages: e253-e253

    • DOI

      10.1038/mtna.2015.28

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Exploring new gene integration sites for gene knock-in by gene-trapping strategy.2015

    • Author(s)
      Nanchi I, Yoshimura Y, Nakamura K, Masago Y, Ohbayashi T, Okuda T.
    • Journal Title

      Transgenic Res

      Volume: 24(3) Issue: 3 Pages: 549-59

    • DOI

      10.1007/s11248-015-9872-x

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Mouse embryonic stem cells with a multi-integrase mouse artificial chromosome for transchromosomic mouse generation.2015

    • Author(s)
      Yoshimura Y, Nakamura K, Endo T, Kajitani N, Kazuki K, Kazuki Y, Kugoh H, Oshimura M, Ohbayashi T.
    • Journal Title

      Transgenic Res.

      Volume: 24(4) Issue: 4 Pages: 717-727

    • DOI

      10.1007/s11248-015-9884-6

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Phenotype specific analyses reveal distinct regulatory mechanism for chronically activated p53.2015

    • Author(s)
      Kirschner K, Samarajiwa SA, Cairns JM, Menon S, Pérez-Mancera PA, Tomimatsu K, Bermejo-Rodriguez C, Ito Y, Chandra T, Narita M, Lyons SK, Lynch AG, Kimura H, Ohbayashi T, Tavaré S, Narita M.
    • Journal Title

      PLoS Genet

      Volume: 11(3)

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Exploring the origin and limitations of kidney regeneration.2015

    • Author(s)
      Endo T, Nakamura J, Sato Y, Asada M, Yamada R, Takase M, Takaori K, Oguchi A, Iguchi T, Higashi AY, Ohbayashi T, Nakamura T, Muso E, Kimura T, Yanagita M.
    • Journal Title

      J Pathol.

      Volume: 236(2) Issue: 2 Pages: 251-63

    • DOI

      10.1002/path.4514

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Dual-color fluorescence imaging to monitor CYP3A4 and CYP3A7 expression in human hepatic carcinoma HepG2 and HepaRG cells.2014

    • Author(s)
      Tsuji S, Kawamura F, Kubiura M, Hayashi A, Ohbayashi T, Kazuki Y, Chesné C, Oshimura M, Tada M.
    • Journal Title

      PLoS One.

      Volume: 9(8) Issue: 8 Pages: e104123-e104123

    • DOI

      10.1371/journal.pone.0104123

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] A novel and stable mouse artificial chromosome vector.2014

    • Author(s)
      Takiguchi M, Kazuki Y, Hiramatsu K, Abe S, Iida Y, Takehara S, Nishida T, Ohbayashi T, Wakayama T, Oshimura M.
    • Journal Title

      ACS Synth Biol

      Volume: 3(12) Issue: 12 Pages: 903-14

    • DOI

      10.1021/sb3000723

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] ゲノムプログラム&コントロールによる新しい遺伝子改変マウス2015

    • Author(s)
      大林徹也
    • Organizer
      第5回 合成生物工学シンポジウム
    • Place of Presentation
      神戸大学統合研究拠点コンベンションホール
    • Year and Date
      2015-08-05
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 人工染色体を用いた蛍光・発光遺伝子導入マウス ES 細胞の樹立法2014

    • Author(s)
      大林 徹也、黒見 靖、吉村 祐貴、中村 和臣、遠藤 猛、中島 芳、近江谷 克裕、 押村 光雄
    • Organizer
      日本実験動物代替法学会 第27回大会
    • Place of Presentation
      横浜国立大学
    • Year and Date
      2014-12-05 – 2014-12-07
    • Related Report
      2014 Research-status Report
  • [Presentation] 人工染色体ベクターによる遺伝子導入細胞を用いた in vitro 安全性試験法の有意性2014

    • Author(s)
      押村 光雄、中島 芳浩、香月 康宏、大林 徹也
    • Organizer
      日本実験動物代替法学会 第27回大会
    • Place of Presentation
      横浜国立大学
    • Year and Date
      2014-12-05 – 2014-12-07
    • Related Report
      2014 Research-status Report
  • [Presentation] 発光・蛍光プローブ遺伝子を導入したマルチカラー細胞/マウスを迅速に作製するシステム

    • Author(s)
      大林徹也
    • Organizer
      光学イメージング2013
    • Place of Presentation
      東京 秋葉原UDX Next1,2
    • Related Report
      2013 Research-status Report
  • [Presentation] マルチインテグレースシステムを用いた遺伝子導入細胞・マウス作製技術

    • Author(s)
      大林徹也
    • Organizer
      第3回ゲノム編集研究会
    • Place of Presentation
      広島 広島大学
    • Related Report
      2013 Research-status Report
  • [Presentation] 発光遺伝子によるマルチレポーターシステムの開発と創薬スクリーニングへの応用

    • Author(s)
      大林徹也
    • Organizer
      日本薬学会第134回年会
    • Place of Presentation
      熊本 鶴屋ホール
    • Related Report
      2013 Research-status Report
    • Invited
  • [Presentation] 人工染色体ベクター導入発光マウス由来細胞を用いた毒性評価システム

    • Author(s)
      大林徹也
    • Organizer
      日本実験動物代替法学会第26回大会
    • Place of Presentation
      京都 京都テレサ
    • Related Report
      2013 Research-status Report
    • Invited
  • [Presentation] マルチインテグレースシステムの開発と染色体工学技術への応用

    • Author(s)
      大林徹也
    • Organizer
      鳥取大学染色体工学研究センター研究成果発表会
    • Place of Presentation
      東京 キャンパス・イノベーションセンター東京
    • Related Report
      2013 Research-status Report
  • [Patent(Industrial Property Rights)] 人工染色体ベクター及び形質転換哺乳類細胞2013

    • Inventor(s)
      中島芳浩、室冨和俊、安永茉由、大林徹也、押村光雄
    • Industrial Property Rights Holder
      中島芳浩、室冨和俊、安永茉由、大林徹也、押村光雄
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2013-263910
    • Filing Date
      2013-12-20
    • Related Report
      2013 Research-status Report

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

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