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Development of homoplasmic mitochondrial genome editing method by using microdevice

Research Project

Project/Area Number 26650068
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Cell biology
Research InstitutionInstitute of Physical and Chemical Research

Principal Investigator

Wada Ken-Ichi  国立研究開発法人理化学研究所, 前田バイオ工学研究室, 協力研究員 (20525919)

Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2014: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywordsミトコンドリアゲノム / マイクロデバイス / 細胞融合 / 細胞質移植 / ゲノム改変
Outline of Final Research Achievements

To carry out functional analysis of mitochondrial (mt) genome, it is necessary to achieve homoplasmic mutation(s) on mt genome, however, feasible mt genome editing method has not been developed. The purpose of this study was to develop a novel mt genome editing method by using a specially-developed microfluidic device which can perform direct cytoplasmic transfer between live single cells via formation of cell fusion through a micro aperture. In the proposed method, homoplasmic mt genome mutation is achieved by transferring of single mitochondrion with mutated mt genome into the mt genome-less cell. In this study, single mitochondrion transfer between single live cells, the most critical matter for achieving homoplasmy of mt genome mutation, was realized by developing a new cell fusion method, in particular, modulating geometry of the microfluidic device.

Report

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

    (19 results)

All 2016 2015 2014

All Journal Article (4 results) (of which Peer Reviewed: 3 results,  Acknowledgement Compliant: 2 results) Presentation (14 results) (of which Int'l Joint Research: 3 results,  Invited: 5 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Effects of ROCK inhibitor Y-27632 on cell fusion through a microslit2015

    • Author(s)
      Ken-Ichi Wada, Kazuo Hosokawa, Yoshihiro Ito, Mizuo Maeda
    • Journal Title

      Biotechnology and Bioengineering

      Volume: 112 Issue: 11 Pages: 2334-2342

    • DOI

      10.1002/bit.25641

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Cell fusion through a microslit between adhered cells and observation of their nuclear behavior2014

    • Author(s)
      Ken-Ichi Wada, Kazuo Hosokawa, Eitaro Kondo, Yoshihiro Ito, Mizuo Maeda
    • Journal Title

      Biotechnology and Bioengineering

      Volume: 111 Issue: 7 Pages: 1464-1468

    • DOI

      10.1002/bit.25190

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Microfluidic perfusion cell culture system confined in 35 mm culture dish for standard biological laboratories2014

    • Author(s)
      Eitaro Kondo, Ken-Ichi Wada, Kazuo Hosokawa, Mizuo Maeda
    • Journal Title

      Journal of Bioscience and Bioengineering

      Volume: 118 Issue: 3 Pages: 356-358

    • DOI

      10.1016/j.jbiosc.2014.02.021

    • NAID

      110009857371

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] マイクロデバイスを用いた細胞操作2014

    • Author(s)
      和田健一
    • Journal Title

      バイオマテリアル-生体材料-

      Volume: 32 Pages: 282-290

    • Related Report
      2014 Research-status Report
  • [Presentation] マイクロ流体デバイスで実現する直接的な細胞質移植技術2016

    • Author(s)
      和田健一、細川和生、伊藤嘉浩、前田瑞夫
    • Organizer
      第25回インテリジェント材料シンポジウム
    • Place of Presentation
      東京都新宿区
    • Year and Date
      2016-01-08
    • Related Report
      2015 Annual Research Report
  • [Presentation] A novel cytoplasm transplantation method by microdevice2015

    • Author(s)
      Ken-Ichi Wada, Kazuo Hosokawa, Yoshihiro Ito, Mizuo Maeda
    • Organizer
      The International Chemical Congress of Pacific Basin Societies (PacifiChem2015)
    • Place of Presentation
      Hawaii, USA
    • Year and Date
      2015-12-15
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] マイクロチップによる高効率な生きた単一細胞間における直接的な細胞質移植2015

    • Author(s)
      和田健一、細川和生、伊藤嘉浩、前田瑞夫
    • Organizer
      第38回日本分子生物学会年会
    • Place of Presentation
      兵庫県神戸市
    • Year and Date
      2015-12-01
    • Related Report
      2015 Annual Research Report
  • [Presentation] On-chip direct cytoplasmic transfer between live single cells2015

    • Author(s)
      Ken-Ichi Wada
    • Organizer
      28th International Microprocess and Nanotechnology Conference
    • Place of Presentation
      Toyama, Japan
    • Year and Date
      2015-11-10
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Prevention of nuclear mixing of fused cells interconnected through a microslit using ROCK inhibitor Y-27632 for direct cytoplasmic transfer between live single cells2015

    • Author(s)
      Ken-Ichi Wada, Kazuo Hosokawa, Yoshihiro Ito, Mizuo Maeda
    • Organizer
      The 19th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2015)
    • Place of Presentation
      Gyeongju, Korea
    • Year and Date
      2015-10-25
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] マイクロデバイスを用いた単一細胞の操作に基づいた細胞生物学の展開2015

    • Author(s)
      和田健一
    • Organizer
      第11回医歯工融合セミナー
    • Place of Presentation
      茨城県つくば市
    • Year and Date
      2015-10-08
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] Selective Organelle Transfer Between Live Single Cells2015

    • Author(s)
      和田健一、細川和生、伊藤嘉浩、前田瑞夫
    • Organizer
      第1回COINS国際シンポジウム
    • Place of Presentation
      東京都 東京大学(伊藤国際学術研究センター・伊藤謝恩ホール)
    • Year and Date
      2015-02-27
    • Related Report
      2014 Research-status Report
  • [Presentation] Selective organelle transplantation between live single cells2015

    • Author(s)
      和田健一、細川和生、伊藤嘉浩、前田瑞夫
    • Organizer
      シングルセルワークショップ
    • Place of Presentation
      神奈川県横浜市(横浜理研)
    • Year and Date
      2015-02-16 – 2015-02-17
    • Related Report
      2014 Research-status Report
  • [Presentation] マイクロデバイスを用いた細胞操作 ―ゲノム操作への応用―2015

    • Author(s)
      和田健一
    • Organizer
      第44回染色体工学研究センターセミナー
    • Place of Presentation
      鳥取県米子市 鳥取大学・とっとりバイオフロンティア1F
    • Year and Date
      2015-01-26
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] マイクロデバイスで迫る細胞のインテリジェンス2015

    • Author(s)
      和田健一
    • Organizer
      第24回インテリジェント材料/システムシンポジウム
    • Place of Presentation
      東京都新宿区(TWIns会議室2F)
    • Year and Date
      2015-01-19
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] マイクロデバイスを用いた選択的な細胞質移植の試み2015

    • Author(s)
      和田健一、細川和生、伊藤嘉浩、前田瑞夫
    • Organizer
      シンポジウム:細胞アッセイの現状と将来
    • Place of Presentation
      東京都目黒区(東京大学生産技術研究所)
    • Year and Date
      2015-01-13
    • Related Report
      2014 Research-status Report
  • [Presentation] マイクロデバイスによる直接的かつ選択的な細胞質移植2014

    • Author(s)
      和田健一、細川和生、伊藤嘉浩、前田瑞夫
    • Organizer
      第24回日本MRS年次大会
    • Place of Presentation
      神奈川県横浜市(開港記念会館)
    • Year and Date
      2014-12-10 – 2014-12-12
    • Related Report
      2014 Research-status Report
  • [Presentation] マイクロデバイスを用いた選択的なオルガネラ移植2014

    • Author(s)
      和田健一、細川和生、前田瑞夫、伊藤嘉浩
    • Organizer
      第37回日本分子生物学会年会
    • Place of Presentation
      神奈川県横浜市(パシフィコ横浜)
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Research-status Report
  • [Presentation] Hippo経路による力学負荷の受容 ―骨格筋肥大機構を視野に入れて―2014

    • Author(s)
      和田健一
    • Organizer
      第6回分子骨格筋代謝研究会
    • Place of Presentation
      京都府京都市 京都大学吉田南キャンパス(人間・環境学研究科棟地階 大講義室)
    • Year and Date
      2014-05-18
    • Related Report
      2014 Research-status Report
    • Invited
  • [Patent(Industrial Property Rights)] 凍結細胞の製造方法2014

    • Inventor(s)
      細川和生、近藤栄太郎、和田健一、前田瑞夫
    • Industrial Property Rights Holder
      細川和生、近藤栄太郎、和田健一、前田瑞夫
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2014-202940
    • Filing Date
      2014-10-01
    • Related Report
      2014 Research-status Report

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

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