An attempt to identify genes involved in PGC specification by using haploid ES cells

• Project/Area Number: 25650077
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Developmental biology
• Research Institution: Kyushu University (2014); Kyoto University (2013)
• Principal Investigator: HAYASHI Katsuhiko 九州大学, 医学(系)研究科(研究院), 教授 (20287486)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 始原生殖細胞 / ES細胞 / 遺伝子スクリーニング / 一倍体 / 順遺伝学的スクリーニング

Outline of Final Research Achievements

Primordial germ cells (PGCs) are origin of all germ cell lineage including sperm and egg. During embryogenesis PGCs are specified at an early developmental stage. This study aims to identify genes involved in PGC specification by using haploid embryonic stem cell lines, which are convenient for forward genetics to identify functional genes. Thus far, we have constructed a plasmid vector that interfere endogenous gene expression at the integrated genomic locus. By transfection of the plasmid vector to haploid ES cells, we have made a pool of ES cell lines in which multiple copies of the vectors are inserted in the genome. Based on the established materials, we are going to screen functional genes involved in PGC specification.

Research Products

• [Journal Article] Hayashi K, Saitou M. (2014)
  • Author(s): Perspectives of germ cell development in vitro in mammals.
  • Journal Title: Animal Science Journal Volume: 85(6) Issue: 6 Pages: 617-626
  • DOI: 10.1111/asj.12199
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Generation of eggs from mouse embryonic stem cells and induced plurip otent stem cells (2013)
  • Author(s): Hayashi K, Saitou M
  • Journal Title: Nature Protocols Volume: 8 Issue: 8 Pages: 1513-1524
  • DOI: 10.1038/nprot.2013.090
  • Peer Reviewed
• [Journal Article] Induction of mouse germ-cell fate by transcription factors in vitro. (2013)
  • Author(s): Nakaki F, Hayashi K, Ohta H, Kurimoto K, Yabuta Y, Saitou M.
  • Journal Title: Nature Volume: 501 Issue: 7466 Pages: 222-226
  • DOI: 10.1038/nature12417
  • Peer Reviewed
• [Journal Article] Stepwise differentiation from naive state pluripotent stem cells to functional primordial germ cells through an epiblast-like state. (2013)
  • Author(s): Hayashi K and Saitou M
  • Journal Title: Methods in molecular biology Volume: 1074 Pages: 175-183
  • DOI: 10.1007/978-1-62703-628-3_13
  • ISBN: 9781627036276, 9781627036283
  • Peer Reviewed
• [Journal Article] Tsix RNA and the germline factor, PRDM14, link X-reactivation and stem cell reprogramming (2013)
  • Author(s): Payer B, Rosenberg M, Yamaji M, Yabuta Y, Michiyo K-A, Hayashi K. Yamanaka S. Saitou M, Lee J. T.
  • Journal Title: Molecular Cell Volume: 52 Issue: 6 Pages: 805-818
  • DOI: 10.1016/j.molcel.2013.10.023
  • Peer Reviewed
• [Journal Article] A mesodermal factor, T, specifies mouse germ cell fate by directly activating germline determinants (2013)
  • Author(s): Aramaki, S., Hayashi, K., Kurimoto, K., Ohta, H., Yabuta, Y., Iwanari, H., Mochizuki, Y., Hamakubo, T., Kato, Y., Shirahige, K., and Saitou, M
  • Journal Title: Developmental Cell Volume: 27 Issue: 5 Pages: 516-529
  • DOI: 10.1016/j.devcel.2013.11.001
  • Peer Reviewed / Open Access
• [Presentation] Propagating generations of healthy mice using manufactured oocytes (2015)
  • Author(s): Katsuhiko Hayashi
  • Organizer: Ovarian Club V
  • Place of Presentation: 香港
  • Year and Date: 2015-01-31 – 2015-02-02
  • Invited
• [Presentation] Generation of eggs from mouse embryonic stem cells (2014)
  • Author(s): Katsuhiko Hayashi
  • Organizer: Society of Reproduction and Fertility
  • Place of Presentation: イギリス
  • Year and Date: 2014-09-21
  • Invited
• [Presentation] In vitro germline formation from pluripotent stem cells (2014)
  • Author(s): 林克彦
  • Organizer: 第４7回日本発生生物学会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-05-27 – 2014-05-30
  • Invited
• [Presentation] Prospects of gamete production from pluripotent stem cells (2014)
  • Author(s): Katsuhiko Hayashi
  • Organizer: The 45th meeting of the Dutch Society of Obstetrics and Gynaecology
  • Place of Presentation: オランダ
  • Year and Date: 2014-05-23
  • Invited
• [Presentation] Making Oocytes and Babies from Stem Cells (2014)
  • Author(s): Katsuhiko Hayashi
  • Organizer: The Congress of the Asia Pacific Initiative on Reproduction
  • Place of Presentation: オーストラリア
  • Year and Date: 2014-04-04 – 2014-04-06
  • Invited
• [Presentation] 生殖細胞系列の体外培養による再構築 (2013)
  • Author(s): 林克彦
  • Organizer: 第25回高遠・分子細胞生物学シンポジウム
  • Place of Presentation: 大津市 延暦寺会館
  • Invited
• [Presentation] Germ cell differentiation from stem cells in mice (2013)
  • Author(s): Katsuhiko Hayashi
  • Organizer: 17th World COngress on IVF
  • Place of Presentation: Tunis, Tunisia
  • Invited
• [Presentation] 始原生殖細胞の分化を体外で再現する培養系の確立と利用 (2013)
  • Author(s): 林克彦
  • Organizer: 第85回日本遺伝学会
  • Place of Presentation: 横浜市 慶應義塾大学
• [Presentation] 生殖細胞系列の再構築培養系の確立と生殖巣の役割 (2013)
  • Author(s): 林克彦
  • Organizer: 第21回日本ステロイドホルモン学会
  • Place of Presentation: 大阪市 千里ライフセンター
  • Invited
• [Book] 実験医学 (2013)
  • Author(s): 林克彦、斎藤通紀
  • Publisher: 羊土社
• [Book] HORMONE FRONTIER IN GYNECOLOGY (2013)
  • Author(s): 林克彦、斎藤通紀
  • Publisher: メディカルレビュー社
• [Book] 産科と婦人科 (2013)
  • Author(s): 林克彦、斎藤通紀
  • Publisher: 診断と治療社