Functional analysis of newly identified DNA demethylation factors in mouse ES cells and its application to somatic cell reprogramming

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K18545
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Developmental biology
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Hatanaka Yuki 国立研究開発法人理化学研究所, バイオリソースセンター, 特別研究員 (70719450)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: 受精卵 / ES細胞 / DNA脱メチル化 / エピジェネティックリプログラミング / エピゲノム

Outline of Final Research Achievements

To explore the application to improvements of somatic nuclear reprogramming, we focused on GSE and H3R17me2a enzyme Mettl23 which are newly identified DNA demethylation factors in mouse zygotes. To this end, we first produced their KO mice and investigated the effect of GSE and Mettl23 deficient on the DNA demethylation in ESCs. The levels of 5mC and its oxidative products were not significantly different between wild type (WT) ESCs and these KO. Interestingly, the level of 5fC was significantly increased in their KOESCs on the promoter regions that undergo DNA demethylation in zygote. The loss of GSE and Mettl23 resulted in the impairment of recruitment of TDG, which excises 5fC and 5CaC. The treatment of TBBD (ellagic acid), which specifically inhibits methylation at H3R17, resulted in the reduction of recruitment of GSE and Histone H3.3 on the target regions. These results suggested that GSE and Mettl23 are responsible for DNA demethylation on defined regions in ESCs.

Free Research Field

分子発生生物学