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- W4214933334 abstract "Enzymatic methylation of cytosine to 5-methylcytosine in DNA is a fundamental epigenetic mechanism involved in mammalian development and disease. DNA methylation is brought about by collective action of three AdoMet-dependent DNA methyltransferases, whose catalytic interactions and temporal interplay are poorly understood. We used structure-guided engineering of the Dnmt1 methyltransferase to enable catalytic transfer of azide tags onto DNA from a synthetic cofactor analog, Ado-6-azide, in vitro. We then CRISPR-edited the Dnmt1 locus in mouse embryonic stem cells to install the engineered codon, which, following pulse internalization of the Ado-6-azide cofactor by electroporation, permitted selective azide tagging of Dnmt1-specific genomic targets in cellulo. The deposited covalent tags were exploited as click handles for reading adjoining sequences and precise genomic mapping of the methylation sites. The proposed approach, Dnmt-TOP-seq, enables high-resolution temporal tracking of the Dnmt1 catalysis in mammalian cells, paving the way to selective studies of other methylation pathways in eukaryotic systems." @default.
- W4214933334 created "2022-03-05" @default.
- W4214933334 creator A5002144881 @default.
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- W4214933334 date "2022-03-01" @default.
- W4214933334 modified "2023-10-01" @default.
- W4214933334 title "Selective chemical tracking of Dnmt1 catalytic activity in live cells" @default.
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- W4214933334 doi "https://doi.org/10.1016/j.molcel.2022.02.008" @default.
- W4214933334 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/35245449" @default.
- W4214933334 hasPublicationYear "2022" @default.
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