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Production of immunodeficient micro-mini pig using genome editing technology.

Research Project

Project/Area Number 18K12062
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 90110:Biomedical engineering-related
Research InstitutionThe University of Tokushima

Principal Investigator

HIRATA Maki  徳島大学, 大学院社会産業理工学研究部(生物資源産業学域), 講師 (10815583)

Co-Investigator(Kenkyū-buntansha) 音井 威重  徳島大学, 大学院社会産業理工学研究部(生物資源産業学域), 教授 (30311814)
谷原 史倫  自治医科大学, 先端医療技術開発センター, 准教授 (90754680)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Keywordsゲノム編集 / ブタ / 免疫不全 / ミニブタ / マイクロミニブタ
Outline of Final Research Achievements

Pigs are useful research model animals that show many characteristics similar to humans. We aimed to produce immunodeficient micro-mini pigs using genome editing technology. In this study, we achieved in vitro fertilization using semen collected from a commercial micro mini pig. In addition, we demonstrated the feasibility of the introduction of double biallelic mutation of the two target genes (IL2RG and GHR), using electroporation to transfer two gRNAs and Cas9 protein to in vitro fertilized embryos. Hence, it will be possible to produce mico-mini-sized immunodeficient pigs by the combination of these technologies.

Academic Significance and Societal Importance of the Research Achievements

近年、iPS細胞をはじめとする幹細胞を用いた再生医療研究は、従来の方法では治療できない難治性疾患への臨床応用にむけ大きな注目を浴びている。本研究はゲノム編集技術を用いて超小型のマイクロミニブタを作出し、多くの研究機関でヒトに近い特徴を有するブタで幹細胞試験を可能にすることを目指した。超小型の免疫不全ブタが活用できれば、多くの研究機関で有用なモデル動物であるブタを用いた研究が可能となり、再生医療研究のみならず、がん研究など多様な研究分野に貢献可能であると思われる。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (15 results)

All 2021 2020 2019 2018

All Journal Article (11 results) (of which Int'l Joint Research: 7 results,  Peer Reviewed: 11 results,  Open Access: 6 results) Presentation (4 results) (of which Int'l Joint Research: 1 results)

  • [Journal Article] Comparison of the effects of introducing the CRISPR/Cas9 system by microinjection and electroporation into porcine embryos at different stages2021

    • Author(s)
      Le AQ., Tanihara F., Wittayarat M., Namula Z., Sato Y., Lin Q., Takebayashi K., Hirata M., Otoi T.
    • Journal Title

      BMC Research Notes

      Volume: 14 Issue: 1 Pages: 7-7

    • DOI

      10.1186/s13104-020-05412-8

    • NAID

      120007147670

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] One-Step Generation of Multiple Gene-Edited Pigs by Electroporation of the CRISPR/Cas9 System into Zygotes to Reduce Xenoantigen Biosynthesis2021

    • Author(s)
      Tanihara F., Hirata M., Nguyen TN., Sawamoto O., Kikuchi T., Otoi T.
    • Journal Title

      International journal of molecular sciences

      Volume: 22 Issue: 5 Pages: 2249-2249

    • DOI

      10.3390/ijms22052249

    • NAID

      120007164926

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Lipofection-Mediated Introduction of CRISPR/Cas9 System into Porcine Oocytes and Embryos2021

    • Author(s)
      Hirata M., Wittayarat M., Namula Z., Le AQ., Lin Q., Takebayashi K., Thongkittidilok C., Tanihara F., Otoi T.
    • Journal Title

      Animals

      Volume: 11 Issue: 2 Pages: 578-578

    • DOI

      10.3390/ani11020578

    • NAID

      120007147677

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Introduction of a point mutation in the KRAS gene of in vitro fertilized porcine zygotes via electroporation of the CRISPR/Cas9 system with single-stranded oligodeoxynucleotides.2021

    • Author(s)
      Wittayarat M., Hirata M., Namula Z., Sato Y., Nguyen TN., Le AQ., Lin Q., Takebayashi K., Tanihara F., Otoi T.
    • Journal Title

      Animal Science Journal,

      Volume: 92 Issue: 1

    • DOI

      10.1111/asj.13534

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] One-step genome editing of porcine zygotes through the electroporation of a CRISPR/Cas9 system with two guide RNAs2020

    • Author(s)
      Hirata M., Wittayarat M., Tanihara F., Sato Y., Namula Z., Le AQ., Lin Q., Takebayashi K., Otoi T.
    • Journal Title

      In Vitro Cellular & Developmental Biology - Animal

      Volume: 56 Issue: 8 Pages: 614-621

    • DOI

      10.1007/s11626-020-00507-9

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Evaluation of multiple gene targeting in porcine embryos by the CRISPR/Cas9 system using electroporation2020

    • Author(s)
      Hirata M., Wittayarat M., Namula Z., Le AQ., Lin Q., Nguyen TN., Takebayashi K., Sato Y., Tanihara F., Otoi T.
    • Journal Title

      Molecular Biology Reports

      Volume: 47 Issue: 7 Pages: 5073-5079

    • DOI

      10.1007/s11033-020-05576-3

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Efficient generation of GGTA1-deficient pigs by electroporation of the CRISPR/Cas9 system into in vitro-fertilized zygotes2020

    • Author(s)
      Tanihara F., Hirata M., Nguyen TN., Sawamoto O., Kikuchi T., Masako Doi., Otoi T.
    • Journal Title

      BMC biotechnology

      Volume: 20 Issue: 1 Pages: 40-40

    • DOI

      10.1186/s12896-020-00638-7

    • NAID

      120007045051

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Generation of CD163-edited pig via electroporation of the CRISPR/Cas9 system into porcine in vitro-fertilized zygotes2020

    • Author(s)
      Tanihara, F., Hirata, M., Nguyen, TN., Le, AQ., Wittayarat, M., Fahrudin, M., Hirano, T., Otoi, T.
    • Journal Title

      Animal Biotechnology

      Volume: - Issue: 2 Pages: 147-154

    • DOI

      10.1080/10495398.2019.1668801

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] The relationship between embryonic development and the efficiency of target mutations in porcine endogenous retroviruses (PERVs) pol genes in porcine embryos2019

    • Author(s)
      Hirata, M., Wittayarat, M., Hirano, T., Nguyen, TN., Le, AQ., Namula, Z., Fahrudin, M., Tanihara, F., Otoi, T.
    • Journal Title

      Animals

      Volume: 9 Issue: 9 Pages: 593-593

    • DOI

      10.3390/ani9090593

    • NAID

      120006877872

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Effects of concentration of CRISPR/Cas9 components on genetic mosaicism in cytoplasmic microinjected porcine embryos2019

    • Author(s)
      Tanihara F, Hirata M, Nguyen NT, LE QA, Hirano T, Otoi T.
    • Journal Title

      Journal of Reproduction and Development

      Volume: 65 Issue: 3 Pages: 209-214

    • DOI

      10.1262/jrd.2018-116

    • NAID

      130007662977

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Relationship among ovarian follicular status, developmental competence of oocytes, and anti-Müllerian hormone levels: A comparative study in Japanese wild boar crossbred gilts and Large White gilts.2019

    • Author(s)
      Tanihara F, Hirata M, Iizuka S, Sairiki S, Nii M, Nguyen NT, Le QA, Hirano T, Otoi T.
    • Journal Title

      Anim Sci J.

      Volume: 印刷中 Issue: 6 Pages: 712-718

    • DOI

      10.1111/asj.13200

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Presentation] ブタ体外受精卵におけるCRISPR/Cas9システムを用いた複数遺伝子の同時改変2019

    • Author(s)
      1.平田真樹、谷原史倫、Nhien Thi Nguyen、Quynh Anh Le、平野隆之、音井威重
    • Organizer
      第7回日本先進医工学ブタ研究会
    • Related Report
      2019 Research-status Report
  • [Presentation] Effects of CRISPR/Cas9-mediated gene targeting of porcine endogenous retrovirus on the developmental competence of porcine embryos2019

    • Author(s)
      Hirata, M., Tanihara, F., Nguyen, TN., Le, AQ., Hirano, T., Otoi, T.
    • Organizer
      The 15th Transgenic Technology Meeting (TT2019)
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research
  • [Presentation] ブタにおけるCrispr/Cas9システムを使用したゲノム編集効率化についての検討2018

    • Author(s)
      平田真樹、谷原史倫、Nhien Thi Nguyen、Zhao Namula、音井威重
    • Organizer
      日本ゲノム編集学会第3回大会
    • Related Report
      2018 Research-status Report
  • [Presentation] ブタにおける成熟卵および1細胞期胚段階でのゲノム編集が胚盤胞の変異導入効率に及ぼす影響2018

    • Author(s)
      平田真樹、谷原史倫、Manita Wittayarat、Nhien Thi Nguyen、 Le Anh Quynh、Zhao Namula、新居雅宏、音井威重
    • Organizer
      第6回日本先進医工学ブタ研究会
    • Related Report
      2018 Research-status Report

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Published: 2018-04-23   Modified: 2022-01-27  

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