Elucidation of the mechanism of maintenance of ES cell pluripotency through analyses of regulatory factors of retroelements

• Project/Area Number: 25640048
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Laboratory animal science
• Research Institution: Nara Medical University
• Principal Investigator: HORIE Kyoji 奈良県立医科大学, 医学部, 教授 (30333446)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2013: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
• Keywords: ES細胞 / レトロエレメント / ゲノム / 遺伝子 / エピジェネティクス

Outline of Final Research Achievements

Analyses of the regulatory mechanisms of ES/iPS cell pluripotency is an important issue for the application of ES/iPS cells toward clinical fields. It has been known that expression of retroelements is tightly regulated in ES/iPS cells. Therefore, aberrant expression of retroelements implies abnormality of pluripotency of ES/iPS cells. We recently developed a method to generate homozygous mutant mouse embryonic stem cells and established a large number of mutant cell lines. in the present study, we screened these homozygous mutant cell lines for abnormal expression of retroelements. Among them, we identified cell lines exhibiting abnormalities in the regulation of pluripotency. It it likely that the mutated genes play important roles in the regulation of the pluripotency or ES/iPS cells.

Research Products

• [Journal Article] Removal of reprogramming transgenes improves the tissue reconstitution potential of keratinocytes generated from human induced pluripotent stem cells (2014)
  • Author(s): Ken Igawaa, Chikara Kokubu, Kosuke Yusa, Kyoji Horie, Yasuhide Yoshimura, Kaori Yamauchi, Hirofumi Suemori, Hiroo Yokozeki, Masashi Toyoda, Nobutaka Kiyokawa, Hajime Okita, Yoshitaka Miyagawa, Hidenori Akutsu, Akihiro Umezawa, Ichiro Katayama, Junji Takeda.
  • Journal Title: Stem Cells Transl Med Volume: 3 Issue: 9 Pages: 992-1001
  • DOI: 10.5966/sctm.2013-0179
  • Peer Reviewed / Open Access
• [Presentation] ホモ変異体マウスES細胞バンクを用いた包括的遺伝子機能解析 (2014)
  • Author(s): 堀江恭二、吉田純子
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
  • Invited
• [Presentation] Rapid identification of homozygous mutant mouse embryonic stem cell clones showing differentiation resistant or differentiation prone phenotype (2014)
  • Author(s): Junko Yoshida, Kyoji Horie
  • Organizer: Global Controls in Stem Cells
  • Place of Presentation: Biopolis, Syngapore
  • Year and Date: 2014-11-05 – 2014-11-07
• [Presentation] Live cell imaging for the analysis of cellular heterogeneity in mouse embryonic stem cell mutant clones identified by forward genetic screening (2014)
  • Author(s): Junko Yoshida, Kyoji Horie
  • Organizer: 12th Annual Meeting of International Society for Stem Cell Research
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2014-06-18 – 2014-06-21
• [Presentation] Genome-wide comparative analyses of retroviral and DNA-type transposon vector integration sites in mouse embryonic stem cells : Implication for reprogramming study (2013)
  • Author(s): Yoshida J, Horie K.
  • Organizer: 11th Annual Meeting of International Society for Stem Cell Research
  • Place of Presentation: Boston, USA