Chemical tracking of DNMT1-specific activity during mESC ground state transition using WGBS

ABSTRACT: The enzymatic methylation of cytosine to 5-methylcytosine in DNA is a crucial epigenetic process implicated in mammalian development and disease. DNA methylation results from the collective performance of three AdoMet-dependent DNA methyltransferases (DNMTs), whose catalytic interplay and temporal dynamics remain poorly elucidated. In this study, we generated a Dnmt1 knock-in mESC line with a N1580A mutation at a single allele, while retaining a functioning wild-type protein. The heterozygous DNMT1 N1580A knock-in line, together with the DNMT1 wild-type, homozygous N1580A knock-in, and knockout mouse embryonic stem cells were subsequently assessed for genome-wide CpG methylation levels using whole-genome bisulfite sequencing. Next, monoallelic DNMT1 N1580A KI cells underwent electroporation using the synthetic cofactor analogue Ado-6-azide and genomic targets unique to DNMT1 were subsequently assessed with Dnmt-TOP-seq. Finally, a comprehensive examination of DNMT1-specific events during mESC ground state transition was conducted. Taken together, the present study paves the way for in vivo monitoring of DNMT1 activity in living tissues in a variety of biological contexts.

ORGANISM(S): Mus musculus

PROVIDER: GSE281223 | GEO | 2026/08/01

REPOSITORIES: GEO