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The human transcription factor occupancy landscape viewed using high-resolution in situ base-conversion strand-specific single-molecule chromatin accessibility mapping

ABSTRACT: Chromatin accessibility profiling is a key tool for mapping the location of cis-regulatory elements (cREs) in the genome and tracking chromatin state dynamics during development, in response to various external and internal stimuli, and in disease contexts. Single-molecule footprinting (SMF) methods that rely on the labeling of individual accessible DNA bases have emerged in recent years as a powerful chromatin accessibility mapping approach, as they provide not just an average readout of accessibility over a given genomic position but also the distribution of accessibility states within a population at the level of individual original DNA molecules. However, SMF approaches have been limited either in their resolution or in labeling readout accuracy. To address these limitations, we have developed a high-resolution strand-specific single molecule footprinting approach (C->U/T-ssSMF) based on the application of highly active sequence context-independent endogenous methylation-insensitive double-strand DNA (dsDNA) deaminases (CseDa01 and LbDa02), which convert accessible cytosines into uracils (and in turn into thymine after amplification). We demonstrate the application of the method to mapping fine-grained single-molecule accessibility states in human cell lines in both a short and a long-read format, and quantifying the occupancy states of individual transcription factors (TFs) as well as TF co-accessibility and strand-specific accessibility patterns.

ORGANISM(S): Homo sapiens

PROVIDER: GSE301497 | GEO | 2025/07/10

REPOSITORIES: GEO

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