Abstract

N 6 -methyladenosine (m 6 A) methylation modification is the most prevalent and abundant internal modification of eukaryotic mRNAs. Increasing evidence has shown that mRNA m 6 A plays important roles in the development of stem cells. However, to the best of our knowledge, no reports about the roles of mRNA m 6 A in mouse female germline stem cells (mFGSCs) have been published. In this study, we compared the genome-wide profiles of mRNA m 6 A methylation and DNA methylation between FGSCs and sandosinbred mice (SIM) embryo-derived thioguanine and ouabain-resistant (STO) cells. qRT-PCR revealed that the expression levels of mRNA m 6 A-related genes ( Mettl3 , Alkbh5 , Ythdf1 , Ythdf2 , Ythdc1 , and Ythdc2 ) in FGSCs were significantly higher than those in STO cells. m 6 A RNA immunoprecipitation sequencing (MeRIP-seq) data further showed that the unique m 6 A-methylated mRNAs in FGSCs and STO cells were related to cell population proliferation and somatic development, respectively. Additionally, knockdown of Ythdf1 inhibited FGSC self-renewal. Comparison of methylated DNA immunoprecipitation sequencing (MeDIP-seq) results between FGSCs and STO cells identified that DNA methylation contributed to FGSC proliferation by suppressing the somatic program. These results suggested that m 6 A regulated FGSC self-renewal possibly through m 6 A binding protein YTHDF1, and DNA methylation repressed somatic programs in FGSCs to maintain FGSC characteristics.