• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Study on practical application of zinc finger nuclease for gene knockout animal production

Research Project

Project/Area Number 25252056
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Integrative animal science
Research InstitutionThe University of Tokyo

Principal Investigator

Naito Kunihiko  東京大学, 大学院農学生命科学研究科(農学部), 教授 (20188858)

Co-Investigator(Renkei-kenkyūsha) Yoshizaki Goro  東京海洋大学, 海洋科学技術研究科, 教授 (70281003)
Kikuchi Kazuhiro  独立行政法人農業生物資源研究所, 動物科学研究領域, 上級研究員 (20360456)
Sugiura Koji  東京大学, 農学生命科学研究科, 准教授 (20595623)
Project Period (FY) 2013-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥46,280,000 (Direct Cost: ¥35,600,000、Indirect Cost: ¥10,680,000)
Fiscal Year 2016: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000)
Fiscal Year 2015: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000)
Fiscal Year 2014: ¥12,610,000 (Direct Cost: ¥9,700,000、Indirect Cost: ¥2,910,000)
Fiscal Year 2013: ¥14,950,000 (Direct Cost: ¥11,500,000、Indirect Cost: ¥3,450,000)
Keywords人工ヌクレアーゼ / CRISPR / ZFN / マウス卵 / ブタ卵 / 魚類卵 / Cas9 / CRISPR/Cas / 魚卵 / 哺乳類卵
Outline of Final Research Achievements

The present study was aimed to promote the practical application of artificial nucleases, which has drawn attention as a new method of gene targeting organism production, and studied various technological developments and application to livestock eggs and fish eggs.
In this study, we showed that triple KO can be prodused at a time using CRISPR/Cas system, and that large-scale mutation can be introduced. As technology development, we constructed offset nicking system which is expected to suppress nonspecific effects and increased availability of original CRISPR/Cas system of other species with different PAM sequence. For the application, we showed the availability of this system to porcine eggs, Yamame and rainbow trout eggs.

Report

(5 results)
  • 2017 Final Research Report ( PDF )
  • 2016 Annual Research Report
  • 2015 Annual Research Report
  • 2014 Annual Research Report
  • 2013 Annual Research Report
  • Research Products

    (32 results)

All 2018 2017 2016 2015 2014 2013

All Journal Article (15 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 14 results,  Open Access: 10 results,  Acknowledgement Compliant: 5 results) Presentation (16 results) (of which Int'l Joint Research: 1 results) Book (1 results)

  • [Journal Article] Contributions of UBE2C and UBE2S to meiotic progression of porcine oocytes2018

    • Author(s)
      Fujioka YA, Onuma A, Fujii W, Sugiura K, Naito K.
    • Journal Title

      Journal of Reproduction and Development

      Volume: 64 Issue: 3 Pages: 253-259

    • DOI

      10.1262/jrd.2018-006

    • NAID

      130007409956

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2016 Annual Research Report
  • [Journal Article] Chd9 mediates highly loosened chromatin structure in growing mouse oocytes2018

    • Author(s)
      Ooga Masatoshi、Funaya Satoshi、Hashioka Yuki、Fujii Wataru、Naito Kunihiko、Suzuki Masataka G.、Aoki Fugaku
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 500 Issue: 3 Pages: 583-588

    • DOI

      10.1016/j.bbrc.2018.04.105

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Efficient mutagenesis by CRISPR/Cas system during meiotic maturation of porcine oocytes2017

    • Author(s)
      Onuma A, Fujii W, Sugiura K, Naito K.
    • Journal Title

      Journal of Reproduction and Development

      Volume: 63 Issue: 1 Pages: 45-50

    • DOI

      10.1262/jrd.2016-094

    • NAID

      130005332294

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Application of dead end-knockout zebrafish to recipients of germ cell transplantation.2017

    • Author(s)
      Li Q, Fujii W, Naito K, Yoshizaki G.
    • Journal Title

      Molecular Reproduction and Development

      Volume: 84 Issue: 10 Pages: 1100-1111

    • DOI

      10.1002/mrd.22870

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Efficient generation of genome-modified mice using Campylobacter jejuni-derived CRISPR/Cas.2017

    • Author(s)
      Fujii W, Ikeda A, Sugiura K, Naito K.
    • Journal Title

      International Jornal of Molecular Sciences

      Volume: 18 Issue: 11 Pages: 2286-2286

    • DOI

      10.3390/ijms18112286

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Zygote-mediated generation of genome-modified mice using Streptococcus thermophiles 1-derived CRISPR/Cas system.2016

    • Author(s)
      Fujii W, Kakuta S, Yoshioka S, Kyuwa S,Sugiura K,Naito K.
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 477 Issue: 3 Pages: 473-476

    • DOI

      10.1016/j.bbrc.2016.06.070

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Analyses of EMI functions on meiotic maturation of porcine oocytes.2016

    • Author(s)
      Fujioka YA, Onuma A, Fujii W, Sugiura K, Naito K.
    • Journal Title

      Molecular Reproduction and Development

      Volume: 83 Issue: 11 Pages: 983-992

    • DOI

      10.1002/mrd.22738

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Development of a mono-promoter-driven CRISPR/Cas9 system in mammalian cells2015

    • Author(s)
      Shin Yoshioka, Wataru Fujii, Tetsuhiro Ogawa, Koji Sugiura, and Kunihiko Naito
    • Journal Title

      Scientific Reports

      Volume: 5 Issue: 1 Pages: 18341-18341

    • DOI

      10.1038/srep18341

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Finding of a highly efficient ZFN pair for <i>Aqpep</i> gene functioning in murine zygotes2015

    • Author(s)
      Fujii W, Onuma A, Yoshioka S, Nagashima K, Sugiura K, Naito K
    • Journal Title

      Journal of Reproduction and Development

      Volume: 61 Issue: 6 Pages: 589-593

    • DOI

      10.1262/jrd.2015-087

    • NAID

      130005113567

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Efficient generation of genome-modified mice via offset-nicking by CRISPR/Cas system.2014

    • Author(s)
      Fujii W, Onuma A, Sugiura K, Naito K.
    • Journal Title

      Biochem Biophys Res Comm

      Volume: 445 Issue: 4 Pages: 791-794

    • DOI

      10.1016/j.bbrc.2014.01.141

    • Related Report
      2014 Annual Research Report 2013 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Nakamura K, Fujii W, Tsuboi M, Tanihata J, Teramoto N, Takeuchi S, Naito K, Yamanouchi K, Nishihara M2014

    • Author(s)
      Nakamura K, Fujii W, Tsuboi M, Tanihata J, Teramoto N, Takeuchi S, Naito K, Yamanouchi K, Nishihara M
    • Journal Title

      Scientific Reports

      Volume: 4 Issue: 1 Pages: 5635-5635

    • DOI

      10.1038/srep05635

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Cytoplasmic Anchoring of cAMP-Dependent Protein Kinase (PKA) by A-Kinase Anchor Proteins (AKAPs) Is Required for Meiotic Arrest of Porcine Full-Grown and Growing Oocytes2014

    • Author(s)
      Nishimura T, Fujii W, Sugiura K, Naito K
    • Journal Title

      Biol Reprod

      Volume: 90 Issue: 3

    • DOI

      10.1095/biolreprod.113.114736

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] One-step Generation of Phenotype-expressing Triple-knockout Mice with Heritable Mutated Alleles by the CRISPR/Cas9 System2014

    • Author(s)
      Fujii W, Onuma A, Sugiura K, Naito K.
    • Journal Title

      Journal of Reproduction and Development

      Volume: 60 Issue: 4 Pages: 324-327

    • DOI

      10.1262/jrd.2013-139

    • NAID

      130004137824

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Repeatable Construction Method for Engineered Zinc Finger Nuclease Based on2013

    • Author(s)
      Wataru Fujii
    • Journal Title

      PloS ONE

      Volume: 8 Issue: 3 Pages: e59801-e59801

    • DOI

      10.1371/journal.pone.0059801

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Efficient generation of large-scale genome-modified-mice using gRNA and CAS9 endonuclease.2013

    • Author(s)
      Fujii W, Kawasaki K, Sugiura K, Naito K.
    • Journal Title

      Nucleic Acids Res.

      Volume: 41 Issue: 20 Pages: e187-e187

    • DOI

      10.1093/nar/gkt772

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Presentation] 生殖細胞欠損ニジマスの作出とその代理親魚への応用2018

    • Author(s)
      藤原亮・片山直人・藤井渉・内藤邦彦・吉崎悟朗
    • Organizer
      平成30年度公益社団法人日本水産学会春季大会
    • Related Report
      2016 Annual Research Report
  • [Presentation] 生殖細胞欠損ヤマメの作出とその代理親魚への応用2017

    • Author(s)
      藤原 亮・片山直人・藤井 渉・内藤邦彦・吉崎悟朗
    • Organizer
      平成29年度公益社団法人日本水産学会春季大会
    • Related Report
      2016 Annual Research Report
  • [Presentation] C. jejuni由来CRISPR/Casによるゲノム改変マウスの作製2017

    • Author(s)
      藤井渉・池田有沙・杉浦幸二・内藤邦彦
    • Organizer
      第123回日本畜産学会
    • Related Report
      2016 Annual Research Report
  • [Presentation] ブタ卵母細胞成熟過程における核外移行因子exportin-1の機能解析2016

    • Author(s)
      小沼あすか・藤井 渉・杉浦幸二・内藤邦彦
    • Organizer
      第121回日本畜産学会
    • Place of Presentation
      日本獣医生命科学大学(東京都武蔵野市)
    • Year and Date
      2016-03-27
    • Related Report
      2015 Annual Research Report
  • [Presentation] Involvement of Chd9 in highly loosened chromatin structure in growing oocytes.2016

    • Author(s)
      Ooga M, Fujii W , Naito K, Aoki F.
    • Organizer
      International Symposium on “Epigenome Dynamics and Regulation in Germ Cells”
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] CRISPR/Casシステムを利用したゲノム改変マウス作製法の開発2015

    • Author(s)
      藤井渉・杉浦幸二・内藤邦彦
    • Organizer
      第16回日本動物遺伝育種学会
    • Place of Presentation
      神戸大学(兵庫県神戸市)
    • Year and Date
      2015-11-06
    • Related Report
      2015 Annual Research Report
  • [Presentation] ドナーDNA条件がCRISPR/Casによるマウス前核期卵のノックインに与える影響2015

    • Author(s)
      永島圭介・藤井 渉・杉浦幸二・内藤邦彦
    • Organizer
      第15回東京大学生命科学シンポジウム
    • Place of Presentation
      東京大学(東京都文京区)
    • Year and Date
      2015-06-27
    • Related Report
      2015 Annual Research Report
  • [Presentation] 生殖細胞欠損サケ科魚類の大量生産を目指したdead end遺伝子のゲノム編集2015

    • Author(s)
      片山直人・定家咲子・矢澤良輔・藤井渉・内藤邦彦・吉崎悟朗
    • Organizer
      第17回マリンバイオテクノロジー学会
    • Place of Presentation
      東京海洋大学(東京都港区)
    • Year and Date
      2015-05-30
    • Related Report
      2015 Annual Research Report
  • [Presentation] ドナーDNA の状態がCRISPR/Casを用いたマウス前核期卵へのノックインに及ぼす影響2015

    • Author(s)
      永島圭介・藤井 渉・杉浦幸二・内藤邦彦
    • Organizer
      第119回日本畜産学会
    • Place of Presentation
      宇都宮大学
    • Year and Date
      2015-03-28 – 2015-03-30
    • Related Report
      2014 Annual Research Report
  • [Presentation] Knockout of dead end gene using zinc finger nuclease in zebrafish2014

    • Author(s)
      Li Q • Fujii W • Naito K • Yoshizaki G
    • Organizer
      平成26年度日本水産学会秋季大会
    • Place of Presentation
      九州大学箱崎キャンパス
    • Year and Date
      2014-09-19 – 2014-09-22
    • Related Report
      2014 Annual Research Report
  • [Presentation] CRISPR/Cas法を用いた筋ジストロフィーモデルラットの作出2014

    • Author(s)
      中村克行・藤井渉・坪井誠也・谷端淳・寺本奈保美・竹内志帆・内藤邦彦・山内啓太郎・西原真杉
    • Organizer
      第157回日本獣医学会
    • Place of Presentation
      北海道大学
    • Year and Date
      2014-09-09 – 2014-09-12
    • Related Report
      2014 Annual Research Report
  • [Presentation] CRISPR/Casシステムを用いたブタ卵成熟過程におけるゲノム改変効率の検討2014

    • Author(s)
      小沼あすか・藤井 渉・杉浦幸二・内藤邦彦
    • Organizer
      第107回日本繁殖生物学会
    • Place of Presentation
      帯広畜産大学
    • Year and Date
      2014-08-20 – 2014-08-24
    • Related Report
      2014 Annual Research Report
  • [Presentation] ブタ卵母細胞の減数分裂過程におけるFBXO5(EMI1)およびFBXO43(EMI2)の関与2014

    • Author(s)
      藤岡良江・藤井 渉・杉浦 幸二・内藤邦彦
    • Organizer
      第107回日本繁殖生物学会
    • Place of Presentation
      帯広畜産大学
    • Year and Date
      2014-08-20 – 2014-08-24
    • Related Report
      2014 Annual Research Report
  • [Presentation] Generation of Dystrophinmutated rats with a CRISPR/Cas system as a new animal model of muscular dystrophy.2014

    • Author(s)
      Nakamura K, Fujii W, Tsuboi M, Tanihata J, Teramoto N, Takeuchi S, Naito K, Yamanouchi K, Nishihara M.
    • Organizer
      2014 FASEB Science Research Conferences
    • Place of Presentation
      Steamboat Springs, Colorado, USA
    • Year and Date
      2014-07-20 – 2014-07-25
    • Related Report
      2014 Annual Research Report
  • [Presentation] オフセットニッキング法によるゲノム改変マウスの作製2014

    • Author(s)
      藤井 渉・小沼 あすか・杉浦 幸二・内藤 邦彦
    • Organizer
      第118回日本畜産学会
    • Place of Presentation
      つくば国際会議場(エポカルつくば)(茨城県つくば市)
    • Related Report
      2013 Annual Research Report
  • [Presentation] 最適化されたCAS9/gRNAシステムによるゲノム改変マウスの作製2013

    • Author(s)
      藤井 渉・川崎 紅・杉浦 幸二・内藤 邦彦
    • Organizer
      第106回日本繁殖生物学会
    • Place of Presentation
      東京農工大学(東京都府中市)
    • Related Report
      2013 Annual Research Report
  • [Book] 哺乳動物の発生工学2014

    • Author(s)
      内藤 邦彦
    • Total Pages
      195
    • Publisher
      朝倉書店
    • Related Report
      2013 Annual Research Report

URL: 

Published: 2013-05-15   Modified: 2019-07-29  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi