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• W3178601136 abstract "Abstract Molecular studies of genome regulation often rely on the ability to map where specific proteins interact with genomic DNA. Existing techniques for mapping protein-DNA interactions genome-wide rely on DNA amplification methods followed by sequencing with short reads, which dissociates joint binding information at neighboring sites, removes endogenous DNA methylation information, and precludes the ability to reliably map interactions in repetitive regions of the genome. To address these limitations, we created a new protein-DNA mapping method, called D irected M ethylation with L ong-read seq uencing (DiMeLo-seq), which methylates DNA near each target protein’s DNA binding site in situ , then leverages the ability to distinguish methylated and unmethylated bases on long, native DNA molecules using long-read, single-molecule sequencing technologies. We demonstrate the optimization and utility of this method by mapping the interaction sites of a variety of different proteins and histone modifications across the human genome, achieving a single-molecule binding site resolution of less than 200 bp. Furthermore, we mapped the positions of the centromeric histone H3 variant CENP-A in repetitive regions that are unmappable with short reads, while simultaneously analyzing endogenous CpG methylation and joint binding events on single molecules. DiMeLo-seq is a versatile method that can provide multimodal and truly genome-wide information for investigating protein-DNA interactions." @default.
• W3178601136 created "2021-07-19" @default.
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• W3178601136 date "2021-07-07" @default.
• W3178601136 modified "2023-09-27" @default.
• W3178601136 title "DiMeLo-seq: a long-read, single-molecule method for mapping protein-DNA interactions genome-wide" @default.
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• W3178601136 doi "https://doi.org/10.1101/2021.07.06.451383" @default.
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