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Establishment of simple culture system for haploid embryonic stem cells

Research Project

Project/Area Number 15K07687
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Animal production science
Research InstitutionGunma University

Principal Investigator

Horii Takuro  群馬大学, 生体調節研究所, 准教授 (00361387)

Co-Investigator(Kenkyū-buntansha) 畑田 出穂  群馬大学, 生体調節研究所, 教授 (50212147)
Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords半数体 / ES細胞 / FACS / CRISPR/Cas
Outline of Final Research Achievements

Haploid cells are useful for studying gene functions because disruption of a single allele can cause loss-of-function phenotypes. Recent success in generating haploid embryonic stem cells (ESCs) in mice, rats, and monkeys provides a new platform for simple genetic manipulation of the mammalian genome. Haploid ESCs tend to diploidize spontaneously due to endoreduplication or missed cytokinesis. It is therefore necessary to purify cells with haploid G1 DNA content at certain intervals. Supported by KAKENHI, we developed a novel haploid purification system that used a scatter plot generated by FACS analysis with a standard 488 nm laser.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (20 results)

All 2017 2016 2015 Other

All Journal Article (7 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 6 results,  Open Access: 4 results,  Acknowledgement Compliant: 2 results) Presentation (9 results) (of which Int'l Joint Research: 2 results,  Invited: 3 results) Book (1 results) Remarks (3 results)

  • [Journal Article] Efficient generation of conditional knockout mice via sequential introduction of lox sites.2017

    • Author(s)
      Horii T, Morita S, Kimura M, Terawaki N, Shibutani M, Hatada I.
    • Journal Title

      Scientific Reports

      Volume: 7 Issue: 1 Pages: 7891-7891

    • DOI

      10.1038/s41598-017-08496-8

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Genome Engineering Using Haploid Embryonic Stem Cells.2017

    • Author(s)
      Horii T, Hatada I.
    • Journal Title

      Prog Mol Biol Transl Sci.

      Volume: 152 Pages: 83-94

    • DOI

      10.1016/bs.pmbts.2017.09.001

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Genome Editing of Mouse by Cytoplasmic Injection.2017

    • Author(s)
      Horii T, Hatada I.
    • Journal Title

      Methods Mol Biol.

      Volume: 1630 Pages: 55-66

    • DOI

      10.1007/978-1-4939-7128-2_5

    • ISBN
      9781493971275, 9781493971282
    • Related Report
      2017 Annual Research Report
  • [Journal Article] Targeted DNA demethylation in vivo using dCas9-peptide repeat and scFv-TET1 catalytic domain fusions.2016

    • Author(s)
      Morita S, Noguchi H, Horii T, Nakabayashi K, Kimura M, Okamura K, Sakai A, Nakashima H, Hata K, Nakashima K, Hatada I
    • Journal Title

      Nat Biotechnol.

      Volume: 34 Issue: 10 Pages: 1060-1065

    • DOI

      10.1038/nbt.3658

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Journal Article] 5-hydroxymethylcytosine marks sites of DNA damage and promotes genome stability.2016

    • Author(s)
      Georgia R. Kafer, Xuan Li, Takuro Horii, Isao Suetake, Shoji Tajima, Izuho Hatada, Peter M. Carlton.
    • Journal Title

      Cell Reports

      Volume: 14 Issue: 6 Pages: 1283-1292

    • DOI

      10.1016/j.celrep.2016.01.035

    • NAID

      120005707793

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Regulation of CpG methylation by Dnmt and Tet in pluripotent stem cells2016

    • Author(s)
      Horii T, Hatada I.
    • Journal Title

      Journal of Reproduction and Development

      Volume: 62 Issue: 4 Pages: 331-335

    • DOI

      10.1262/jrd.2016-046

    • NAID

      130005406815

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Genome Editing Using Mammalian Haploid Cells2015

    • Author(s)
      Horii T, Hatada I
    • Journal Title

      International Journal of Molecular Science

      Volume: 16 Issue: 10 Pages: 23604-23614

    • DOI

      10.3390/ijms161023604

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] Targeted manipulation of epigenome using CRISPR/Cas92017

    • Author(s)
      Horii T.
    • Organizer
      4th World Congress of Reproductive Biology
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] CRISPR/Casによるゲノム編集技術の基礎と応用2017

    • Author(s)
      堀居拓郎
    • Organizer
      第42回組織細胞化学講習会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] Lox配列の2ステップ導入による効率的なコンディショナルノックアウトマウスの作製2017

    • Author(s)
      堀居拓郎、森田純代、木村美香、寺脇直美、渋谷海大、畑田出穂
    • Organizer
      日本分子生物学会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Cas9タンパクおよびエレクトロポレーション法を用いたコンディショナルノックアウトマウスの作製2016

    • Author(s)
      堀居拓郎、森田純代、木村美香、寺脇直美、畑田出穂
    • Organizer
      第1回日本ゲノム編集学会
    • Place of Presentation
      広島
    • Year and Date
      2016-09-06
    • Related Report
      2016 Research-status Report
  • [Presentation] ES細胞の多能性にはTetによるNr2f2プロモーター領域の脱メチル化が必要であ2016

    • Author(s)
      堀居拓郎、森田純代、木村美香、寺脇直美、木村博信、末武勲、田嶋正二、安部由美子、畑田出穂
    • Organizer
      第10回日本エピジェネティクス研究会年会
    • Place of Presentation
      大阪
    • Year and Date
      2016-05-19
    • Related Report
      2016 Research-status Report
  • [Presentation] Tetタンパクは転写因子Nr2f2のプロモーターを脱メチル化することによりES細胞の多能性を維持する2016

    • Author(s)
      堀居拓郎、森田純代、木村美香、寺脇直美、木村博信、末武勲、田嶋正二、安部由美子、畑田出穂
    • Organizer
      第109回日本繁殖生物学会年会
    • Place of Presentation
      相模原
    • Related Report
      2016 Research-status Report
  • [Presentation] Tet遺伝子シングル、ダブルおよびトリプルノックアウトES細胞株の特性2015

    • Author(s)
      堀居拓郎、森田純代、木村美香、小林遼平、田村大樹、木村博信、末武勲、田嶋正二、安部由美子、畑田出穂
    • Organizer
      第38回日本分子生物学会年会
    • Place of Presentation
      神戸国際展示場(兵庫県神戸市)
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report
  • [Presentation] ES細胞の初期分化にはTetによるNr2f2プロモーター領域の脱メチル化が必要である2015

    • Author(s)
      堀居拓郎、森田純代、木村美香、小林遼平、田村大樹、木村博信、末武勲、田嶋正二、安部由美子、畑田出穂
    • Organizer
      第9回日本エピジェネティクス研究会年会
    • Place of Presentation
      学術総合センター 一橋講堂(東京都千代田区)
    • Year and Date
      2015-05-25
    • Related Report
      2015 Research-status Report
  • [Presentation] 卵細胞を用いたゲノム編集の現状と利用2015

    • Author(s)
      堀居拓郎
    • Organizer
      第60回日本生殖医学会
    • Place of Presentation
      パシフィコ横浜(神奈川県横浜市)
    • Year and Date
      2015-04-27
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research / Invited
  • [Book] All About ゲノム編集2016

    • Author(s)
      畑田出穂、森田純代、堀居拓郎
    • Total Pages
      240
    • Publisher
      羊土社
    • Related Report
      2016 Research-status Report
  • [Remarks] Hatada Laboratory

    • URL

      http://epigenome.dept.showa.gunma-u.ac.jp/~hatada/index.php

    • Related Report
      2017 Annual Research Report
  • [Remarks] 群馬大学生体調節研究所 ゲノム科学リソース分野(畑田研究室)

    • URL

      http://epigenome.dept.showa.gunma-u.ac.jp/~hatada/

    • Related Report
      2016 Research-status Report
  • [Remarks] 群馬大学 生体調節研究所 生体情報ゲノムリソースセンター ゲノム科学リソース分野

    • URL

      http://epigenome.dept.showa.gunma-u.ac.jp/~hatada/

    • Related Report
      2015 Research-status Report

URL: 

Published: 2015-04-16   Modified: 2019-03-29  

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