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A gene expression control system using a genome exchange technology on a mammalian artificial chromosome vector

Research Project

Project/Area Number 15K12141
Research Category

Grant-in-Aid for Challenging Exploratory Research

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

Principal Investigator

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

Co-Investigator(Kenkyū-buntansha) 古倉 健嗣  鳥取大学, 医学部, 助教 (30344039)
中村 和臣  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) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2016: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Keywords人工染色体ベクター / 遺伝子工学 / 遺伝子改変技術 / ゲノム改変技術 / ゲノム編集 / インテグレース / システム生物学 / 合成生物学
Outline of Final Research Achievements

Mammalian artificial chromosome (MAC) vectors have several advantages as gene delivery vectors, such as stable and independent maintenance in host cells without integration
Previously, a MAC containing a multi-integrase platform (MI-MAC) was developed to facilitate the transfer of multiple genes into desired cells. To introduce multiple-genes into MI-MAC platform, we developed multiple-gene-loading method by combining multi-integration system-equipped MAC vector and CRISPR-Cas9. Next, we developed multiple expression cassette exchange system via three integrase systems on a MAC vector. We designed three excision cassettes that expressed luciferase, where the luciferase expression could be exchanged to a fluorescent protein by site-specific recombination. The transient expression of an integrase caused the targeted luciferase activity to be lost and fluorescence was activated. Furthermore, we developed in vitro cisplatin nephrotoxicity test using these systems.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (8 results)

All 2018 2017 2015

All Journal Article (7 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 7 results,  Open Access: 5 results,  Acknowledgement Compliant: 2 results) Presentation (1 results) (of which Invited: 1 results)

  • [Journal Article] Development of a multiple-gene-loading method by combining multi-integration system-equipped mouse artificial chromosome vector and CRISPR-Cas92018

    • Author(s)
      Honma Kazuhisa、Abe Satoshi、Endo Takeshi、Uno Narumi、Oshimura Mitsuo、Ohbayashi Tetsuya、Kazuki Yasuhiro
    • Journal Title

      PLOS ONE

      Volume: 13 Issue: 3 Pages: e0193642-e0193642

    • DOI

      10.1371/journal.pone.0193642

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Multiple expression cassette exchange via TP901-1, R4, and Bxb1 integrase systems on a mouse artificial chromosome.2017

    • Author(s)
      Tomimatsu K, Kokura K, Nishida T, Yoshimura Y, Kazuki Y, Narita M, Oshimura M, Ohbayashi T.
    • Journal Title

      FEBS Open Bio.

      Volume: 7 Issue: 3 Pages: 306-317

    • DOI

      10.1002/2211-5463.12169

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [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 Research-status 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 Research-status 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 Research-status 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 Issue: 3 Pages: e1005053-e1005053

    • DOI

      10.1371/journal.pgen.1005053

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [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 Research-status Report
    • Peer Reviewed
  • [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 Research-status Report
    • Invited

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Published: 2015-04-16   Modified: 2019-03-29  

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