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Genome-wide profiling of DNA methyltransferases in mammalian cells

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Abstract

Chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) is currently the method of choice to determine binding sites of chromatin-associated factors in a genome-wide manner. Here, we describe a method to investigate the binding preferences of mammalian DNA methyltransferases (DNMT) based on ChIP-seq using biotin-tagging. Stringent ChIP of DNMT proteins based on the strong interaction between biotin and avidin circumvents limitations arising from low antibody specificity and ensures reproducible enrichment. DNMT-bound DNA fragments are ligated to sequencing adaptors, amplified and sequenced on a high-throughput sequencing instrument. Bioinformatic analysis gives valuable information about the binding preferences of DNMTs genome-wide and around promoter regions. This method is unconventional due to the use of genetically engineered cells; however, it allows specific and reliable determination of DNMT binding.

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Manzo, M., Ambrosi, C., & Baubec, T. (2018). Genome-wide profiling of DNA methyltransferases in mammalian cells. In Methods in Molecular Biology (Vol. 1766, pp. 157–174). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7768-0_9

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