Comprehensive analysis of genome imprinting regulation

• Project/Area Number: 15K18458
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Genome biology
• Research Institution: Kitasato University
• Principal Investigator: SEKITA YOICHI 北里大学, 理学部, 准教授 (20431950)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: エピジェネティクス / ゲノムインプリンティング / enChIP / CRISPR/Cas9 / 幹細胞 / DNAメチル化 / エピゲノム / クロマチンリモデリング因子 / インプリンティング疾患

Outline of Final Research Achievements

Epigenetic gene regulation has critical roles for various physiological phenomena including development and diseases. Genomic imprinting is a typical epigenetic mechanism by which imprinted genes are expressed in a parent-of-origin specific manner. In this study, I searched for protein factors involved in imprinting regulation by combining enChIP, a new strategy developed from the CRISPR/Cas9 system, and special cell lines produced by nuclear transfer technology of fertilized egg. A lenti virus vector that expresses PA-tagged dCas9 for enChIP experiments was constructed and has been deposited to RIKEN BRC and Addgene. By using this construct, I identified proteins that bind to imprinting control regions in a DNA methylation dependent manner. It is suggested that one of these factors is possibly involved in a DNA methylation control mechanism.

Research Products

• [Journal Article] Reprogramming of germ cells into pluripotency (2016)
  • Author(s): Sekita Y, Nakamura T, Kimura T
  • Journal Title: World J. Stem Cells Volume: 8 Issue: 8 Pages: 251-259
  • DOI: 10.4252/wjsc.v8.i8.251
  • Peer Reviewed / Open Access
• [Journal Article] Stella controls chromocenter formation through regulation of Daxx expression in 2-cell embryos. (2015)
  • Author(s): Arakawa, T., Nakatani, T., Oda, M., Kimura, Y., Sekita, Y., Kimura, T., Nakamura, T., and Nakano, T.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 466 Issue: 1 Pages: 60-60
  • DOI: 10.1016/j.bbrc.2015.08.106
  • Peer Reviewed / Open Access
• [Journal Article] CRISPR/Cas9-mediated reporter knock-in in mouse haploid embryonic stem cells (2015)
  • Author(s): Kimura, Y., Oda, M., Nakatani, T., Sekita, Y., Monfort, A., Wutz, A., Mochizuki, H., and Nakano, T
  • Journal Title: Scientific Reports Volume: 5 Issue: 1 Pages: 10710-10710
  • DOI: 10.1038/srep10710
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Stella preserves maternal chromosome integrity by inhibiting 5hmC-dependent gammaH2AX accumulation. (2015)
  • Author(s): Nakatani et al.
  • Journal Title: EMBO Reports Volume: 16 Issue: 5 Pages: 582-589
  • DOI: 10.15252/embr.201439427
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] AKT promotes somatic cell nuclear reprogramming through α-KG (2018)
  • Author(s): 関田洋一、松元愛香里、川崎佑季、杉浦悠毅、紺野亮、小寺義男、幸田尚、石 野史敏、木村透
  • Organizer: 第16回幹細胞シンポジウム
• [Presentation] AKT promotes somatic cell nuclear reprogramming through α-KG (2017)
  • Author(s): 松元愛香里、関田洋一、川崎佑季、杉浦悠毅、小寺義男、幸田尚、石野史敏、木村透
  • Organizer: 第15回幹細胞シンポジウム
  • Place of Presentation: 東京大学（東京都文京区）
  • Year and Date: 2017-05-26
• [Presentation] Identification and analysis of imprinting control region-binding proteins (2017)
  • Author(s): 関田洋一、吉村祐樹、藤井穂高、木村透
  • Organizer: 第15回幹細胞シンポジウム
• [Presentation] AKT promotes somatic cell nuclear reprogramming through α-KG (2017)
  • Author(s): 松元愛香里、関田洋一、川崎佑季、杉浦悠毅、小寺義男、幸田尚、石 野史敏、木村透
  • Organizer: 第15回幹細胞シンポジウム
• [Presentation] Essential roles of Snf5 in female germ cell sex differentiation (2017)
  • Author(s): 伊藤駿瑛、関田洋一、Charles Roberts、木村透
  • Organizer: 第15回幹細胞シンポジウム
• [Presentation] Physiological roles of the genes expressed during maternal zygotic transition (2017)
  • Author(s): 神﨑理子、若林美月、高田修二、関田洋一、木村透
  • Organizer: 第15回幹細胞シンポジウム
• [Presentation] enChIP法によるゲノムインプリンティング制御領域結合因子の網羅的探索 (2016)
  • Author(s): 吉村祐樹、関田洋一、鏡雅代、紺野亮、藤井穂高、小寺義男、木村透
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-11-30
• [Presentation] AKTシグナルはαケトグルタル酸を介して体細胞核の初期化を促進する (2016)
  • Author(s): 松元愛香里、関田洋一、川崎佑季、杉浦悠毅、小寺義男、幸田尚、石野史敏、木村透
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-11-30
• [Presentation] 細胞の初期化過程でのシグナル伝達経路の役割 (2015)
  • Author(s): 関田洋一、木村透
  • Organizer: 2015年分子生物学会
  • Place of Presentation: 神戸国際会議場（兵庫県神戸市）
  • Year and Date: 2015-12-03