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Development of genetically modified microminipigs that can contribute to cancer treatments as disease model animals

Research Project

Project/Area Number 16K08085
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Integrative animal science
Research InstitutionKagoshima University

Principal Investigator

MIYOSHI Kazuchika  鹿児島大学, 農水産獣医学域農学系, 教授 (70363611)

Co-Investigator(Kenkyū-buntansha) 佐藤 正宏  鹿児島大学, 医用ミニブタ・先端医療開発研究センター, 教授 (30287099)
川口 博明  鹿児島大学, 医歯学域医学系, 准教授 (60325777)
Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywords体細胞核移植 / 疾患モデル動物 / マイクロミニピッグ
Outline of Final Research Achievements

Treatment with valproic acid after activation enhanced the blastocyst formation rate of microminipig cloned embryos derived from somatic cells in which specific genes were disrupted. In addition, specific gene-disrupted microminipig blastocysts were obtained by electroporation of CRISPR/Cas9 genome editing components into somatic cell nuclear transfer embryos. On the basis of these results, an efficient protocol for the production of specific gene-disrupted microminipig embryos was established.

Academic Significance and Societal Importance of the Research Achievements

ブタは、解剖学的・生理学的にヒトとの類似点が多いので、目的に応じて遺伝子を改変すればヒト疾患モデル動物として有用である。しかし、食用ブタやミニブタの飼育管理には広いスペースや多大な労力・コストが必要となるので、利用できる施設は限られてしまう。一方、マイクロミニピッグであれば、成体重が10kg以下なので多くの施設で利用できる。本研究の成果は、標的遺伝子を破壊したマイクロミニピッグの作出につながる。その結果、多くの施設で利用可能なヒト疾患モデル動物を作出できるようになり、それを用いて医薬品の開発や病気治療法の確立が進むと考えられる。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (14 results)

All 2018 2017 2016 Other

All Journal Article (9 results) (of which Peer Reviewed: 9 results,  Open Access: 5 results) Presentation (4 results) (of which Invited: 1 results) Remarks (1 results)

  • [Journal Article] The Combinational Use of CRISPR/Cas9 and Targeted Toxin Technology Enables Efficient Isolation of Bi-Allelic Knockout Non-Human Mammalian Clones2018

    • Author(s)
      Watanabe Satoshi、Sakurai Takayuki、Nakamura Shingo、Miyoshi Kazuchika、Sato Masahiro
    • Journal Title

      International Journal of Molecular Sciences

      Volume: 19 Issue: 4 Pages: 1075-1075

    • DOI

      10.3390/ijms19041075

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Timing of CRISPR/Cas9-related mRNA microinjection after activation as an important factor affecting genome editing efficiency in porcine oocytes2018

    • Author(s)
      Sato M, Maeda K, Koriyama M, Inada E, Saitoh I, Ohtsuka M, Nakamura S, Sakurai, T, Watanabe S, Miyoshi K
    • Journal Title

      Theriogenology

      Volume: 108 Pages: 29-38

    • DOI

      10.1016/j.theriogenology.2017.11.030

    • NAID

      120007099790

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Selection of human blastocysts with a high implantation potential based on timely compaction2017

    • Author(s)
      Mizobe Y, Ezono Y, Tokunaga M, Oya N, Iwakiri R, Yoshida N, Sato Y, Onoue N, Miyoshi K
    • Journal Title

      Journal of Assisted Reproduction and Genetics

      Volume: 34 Issue: 8 Pages: 991-997

    • DOI

      10.1007/s10815-017-0962-y

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Efficient Generation of Somatic Cell Nuclear Transfer-Competent Porcine Cells with Mutated Alleles at Multiple Target Loci by Using CRISPR/Cas9 Combined with Targeted Toxin-Based Selection System.2017

    • Author(s)
      Sato M, Miyoshi K, Nakamura S, Ohtsuka M, Sakurai T, Watanabe S, Kawaguchi H, Tanimoto A.
    • Journal Title

      Int J Mol Sci

      Volume: 18 Issue: 12 Pages: 2610-2610

    • DOI

      10.3390/ijms18122610

    • NAID

      120007099791

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Development of a culture medium for rat 1-cell embryos2016

    • Author(s)
      Miyoshi K
    • Journal Title

      Journal of Mammalian Ova Research

      Volume: 33 Issue: 1 Pages: 11-16

    • DOI

      10.1274/jmor.33.11

    • NAID

      40020876145

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Journal Article] Effects of early cleavage patterns of human embryos on subsequent in vitro development and implantation2016

    • Author(s)
      Mizobe Y, Oya N, Iwakiri R, Yoshida N, Sato Y, Miyoshi K, Tokunaga M, Ezono Y
    • Journal Title

      Fertility and Sterility

      Volume: 106 Issue: 2 Pages: 348-353

    • DOI

      10.1016/j.fertnstert.2016.04.020

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Journal Article] The piggyBac-Based Gene Delivery System Can Confer Successful Production of Cloned Porcine Blastocysts with Multigene Constructs.2016

    • Author(s)
      Sato M, Maeda K, Koriyama M, Inada E, Saitoh I, Miura H, Ohtsuka M, Nakamura S, Sakurai T, Watanabe S, Miyoshi K.
    • Journal Title

      Int J Mol Sci.

      Volume: 17 Issue: 9 Pages: 1424-1424

    • DOI

      10.3390/ijms17091424

    • NAID

      130007023583

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Birth of Cloned Microminipigs Derived from Somatic Cell Nuclear Transfer Embryos that have been Transiently Treated with Valproic Acid.2016

    • Author(s)
      Miyoshi K, Maeda K, Akioka K, Sato M, Kawaguchi H, Tanimoto A
    • Journal Title

      Cellular Reprogramming

      Volume: 6 Issue: 6 Pages: 390-400

    • DOI

      10.1089/cell.2016.0025

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Developmental ability of embryos produced from oocytes with fragile oolemma by intracytoplasmic sperm injection2016

    • Author(s)
      Mizobe Y, Oya N, Iwakiri R, Yoshida N, Sato Y, Onoue N, Miyoshi K, Tokunaga M, Ezono Y
    • Journal Title

      Journal of Assisted Reproduction and Genetics

      Volume: 33 Issue: 12 Pages: 1685-1690

    • DOI

      10.1007/s10815-016-0811-4

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Presentation] マイクロミニピッグにおける体細胞クローニング技術の確立およびそれを用いた遺伝子ノックアウト動物作出の試み2018

    • Author(s)
      三好和睦
    • Organizer
      第20回応用薬理シンポジウム
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 体細胞核移植胚への直接electroporation(GENTEP)は効率的なゲノム編集ブタ作製に有効である2018

    • Author(s)
      佐藤正宏, 三好和睦, 松永将伍, 渡部聡, 中村伸吾, 川口博明, 谷本昭英
    • Organizer
      第41回日本分子生物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] piggyBac遺伝子導入系により複数遺伝子が同時導入されたブタ細胞は核移植後のクローン胚の発生を保証する2016

    • Author(s)
      佐藤正宏, 前田昂亮, 郡山実優, 稲田絵美, 齋藤一誠, 大塚正人, 中村伸吾, 桜井敬之, 渡部聡, 三好和睦
    • Organizer
      第109回日本繁殖生物学会
    • Place of Presentation
      麻布大学(神奈川県相模原市)
    • Year and Date
      2016-09-12
    • Related Report
      2016 Research-status Report
  • [Presentation] ヒトアポリポプロテイン(a)発現ミニブタの作出2016

    • Author(s)
      小沢政之, 大久津昌治, 三好和睦
    • Organizer
      第109回日本繁殖生物学会
    • Place of Presentation
      麻布大学(神奈川県相模原市)
    • Year and Date
      2016-09-12
    • Related Report
      2016 Research-status Report
  • [Remarks] 鹿児島大学農学部農業生産科学科畜産科学コース家畜繁殖学研究室ホームページ

    • URL

      http://ace1.agri.kagoshima-u.ac.jp/agri0015/

    • Related Report
      2018 Annual Research Report 2017 Research-status Report 2016 Research-status Report

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Published: 2016-04-21   Modified: 2020-03-30  

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