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Development of a multiple knockout technology with CRISPR/Cas9 in rats

Research Project

Project/Area Number 25890011
Research Category

Grant-in-Aid for Research Activity Start-up

Allocation TypeSingle-year Grants
Research Field Laboratory animal science
Research InstitutionKyoto University

Principal Investigator

YOSHIMI Kazuto  京都大学, 医学(系)研究科(研究院), 研究員 (50709813)

Project Period (FY) 2013-08-30 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywordsラット / ゲノム編集 / ノックアウト / ノックイン / CRISPR/Cas9 / CRISPR/Cas / チロシナーゼ
Outline of Final Research Achievements

Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system was applied as a newly and efficient genome editing tool in rats. We focused on the coat color related gene, Tyr, Asip and Kit genes. Microinjection of guide RNA (gRNA) and Cas9 mRNA into rat embryos produced a variety of insertion or deletion mutations and generated knockout (KO) rats by using the CRISPR/Cas9 system. Furthermore, combination of CRISPR/Cas9 and single-stranded oligodeoxyribonucleotides (ssODNs) enabled us to generate several types of targeted knock-in (KI) rats, such as SNP substitution, short DNA fragment integration, and large DNA fragment elimination. Therefore, CRISPR/Cas9 provides an efficient and robust genome engineering tool to generate multiple KO and KI rats.

Report

(3 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Annual Research Report
  • Research Products

    (18 results)

All 2015 2014 2013 Other

All Journal Article (3 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results,  Acknowledgement Compliant: 1 results) Presentation (12 results) (of which Invited: 2 results) Book (1 results) Remarks (1 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Allele-specific genome editing and correction of disease-associated phenotypes in rats using the CRISPR-Cas platform.2014

    • Author(s)
      Kazuto Yoshimi, Takehito Kaneko, Birger Voigt, Tomoji Mashimo
    • Journal Title

      Nature Communications

      Volume: 5 Issue: 1 Pages: 4240-4240

    • DOI

      10.1038/ncomms5240

    • NAID

      120005451811

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] ラットにおけるゲノム編集技術革命(Progress of genome editing technologies in rats)2014

    • Author(s)
      吉見一人、金子武人、真下知士
    • Journal Title

      実験医学

      Volume: 32 (11) Pages: 1715-1720

    • Related Report
      2014 Annual Research Report
  • [Journal Article] ラットにおけるTALENおよびCRISPR/Cas9を用いた遺伝子改変2013

    • Author(s)
      吉見一人、金子武人、真下知士
    • Journal Title

      実験医学別冊 最強のステップアップシリーズ「今すぐ始めるゲノム編集」(山本卓/編)

      Volume: - Pages: 109-119

    • Related Report
      2013 Annual Research Report
  • [Presentation] Improved Protocols for Generating Genetically-modified Rats with the CRISPR/Cas9 System2015

    • Author(s)
      Kazuto Yoshimi, Takehito Kaneko, Birger Voigt, Tomoji Mashimo
    • Organizer
      BRI International Symposium 2015
    • Place of Presentation
      新潟
    • Year and Date
      2015-03-05 – 2015-03-06
    • Related Report
      2014 Annual Research Report
  • [Presentation] Improved Protocols for Generating Genetically-modified Rats with the CRISPR/Cas9 System2014

    • Author(s)
      Kazuto Yoshimi, Takehito Kaneko, Birger Voigt, Tomoji Mashimo
    • Organizer
      2014 Rat Genomics & Models
    • Place of Presentation
      Hinxton, UK
    • Year and Date
      2014-12-01 – 2014-12-04
    • Related Report
      2014 Annual Research Report
  • [Presentation] CRISPR/Casシステムによるノックインラットの作製法 (Efficient generation of knock-in rats using CRISPR/Cas system)2014

    • Author(s)
      吉見一人、金子武人、真下知士
    • Organizer
      第37回日本分子生物学会総会
    • Place of Presentation
      横浜
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Annual Research Report
  • [Presentation] CRISPR/Cas9を利用した効率的な遺伝子改変ラット作製法2014

    • Author(s)
      吉見一人、国広弥生、林真智子、服部晃佑、金子武人、真下知士
    • Organizer
      第4回ゲノム編集研究会
    • Place of Presentation
      広島
    • Year and Date
      2014-10-06 – 2014-10-07
    • Related Report
      2014 Annual Research Report
  • [Presentation] ゲノム編集技術がもたらす実験動物の新展開 ~ラットを中心に~2014

    • Author(s)
      吉見一人
    • Organizer
      第31回藤田カンファレンス
    • Place of Presentation
      大阪
    • Year and Date
      2014-09-20 – 2014-09-21
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] Allele-specific genome editing and correction of coat color-related phenotypes in rats using the CRISPR/Cas system2014

    • Author(s)
      Kazuto Yoshimi, Takehito Kaneko, Birger Voigt, Tomoji Mashimo
    • Organizer
      Cold Spring Harbor Asia Conference “Systems Biology of Gene Regulation and Genome Editing”
    • Place of Presentation
      Suzhou, China
    • Year and Date
      2014-09-09 – 2014-09-12
    • Related Report
      2014 Annual Research Report
  • [Presentation] Allele-specific genome editing and correction of coat color-related phenotypes in rats using CRISPR/Cas92014

    • Author(s)
      Kazuto Yoshimi
    • Organizer
      5th Young EURAT Investigators Symposium
    • Place of Presentation
      Malaga, Spain
    • Year and Date
      2014-06-05 – 2014-06-06
    • Related Report
      2014 Annual Research Report
  • [Presentation] CRISPR/Casを用いたノックインラットの作製2014

    • Author(s)
      吉見一人、金子武人、真下知士
    • Organizer
      第61回日本実験動物学会総会
    • Place of Presentation
      札幌
    • Year and Date
      2014-05-15 – 2014-05-17
    • Related Report
      2014 Annual Research Report
  • [Presentation] CRISPR/Casシステム:ラット遺伝子改変技術のさらなる展開2014

    • Author(s)
      吉見一人
    • Organizer
      第7回ラットリソースリサーチ研究会
    • Place of Presentation
      京都大学、京都
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] CRISPR/Casを用いたSNP特異的遺伝子改変ラットの作製2013

    • Author(s)
      吉見一人、金子武人、芹川忠夫、真下知士
    • Organizer
      第3回ゲノム編集研究会
    • Place of Presentation
      広島大学、東広島
    • Related Report
      2013 Annual Research Report
  • [Presentation] Knockout and knock-in mutations for rat albino gene generated by CRISPR/Cas2013

    • Author(s)
      Kazuto Yoshimi, Takehito Kaneko, Tomoji Mashimo
    • Organizer
      2013 Rat Genomics & Models Meeting
    • Place of Presentation
      Cold Spring Harbor, NY, USA
    • Related Report
      2013 Annual Research Report
  • [Presentation] Gene targeting technologies in rats: ZFN, TALEN, and CRISPR2013

    • Author(s)
      Tomoji Mashimo, Kazuto Yoshimi, Takehito Kaneko, Tadao Serikawa
    • Organizer
      2013 Rat Genomics & Models Meeting
    • Place of Presentation
      Cold Spring Harbor, NY, USA
    • Related Report
      2013 Annual Research Report
  • [Book] Targeted Genome Editing Using Site-Specific Nucleases2015

    • Author(s)
      Takashi Yamamoto, Hiroshi Ochiai, Tetsushi Sakuma, Kazuto Yoshimi, Takehito Kaneko, Birger Voigt, Tomoji Mashimo 他
    • Total Pages
      205
    • Publisher
      Springer Japan
    • Related Report
      2014 Annual Research Report
  • [Remarks] CRISPR/Casシステムを用いたアレル特異的ゲノム編集とラット毛色突然変異の修復

    • URL

      http://www.kyoto-u.ac.jp/ja/research/research_results/2014/140626_1.html

    • Related Report
      2014 Annual Research Report
  • [Patent(Industrial Property Rights)] 哺乳動物の標的ゲノム領域にDNAをノックインする方法及び細胞2014

    • Inventor(s)
      真下知士、吉見一人、金子武人
    • Industrial Property Rights Holder
      真下知士、吉見一人、金子武人
    • Industrial Property Rights Type
      特許
    • Filing Date
      2014-11-20
    • Related Report
      2014 Annual Research Report

URL: 

Published: 2013-09-12   Modified: 2019-07-29  

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