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Long-range single-molecule mapping of chromatin accessibility in eukaryotes
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Long-range single-molecule mapping of chromatin accessibility in eukaryotes
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Long-range single-molecule mapping of chromatin accessibility in eukaryotes
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Journal Article
Long-range single-molecule mapping of chromatin accessibility in eukaryotes
2020
Overview
Mapping open chromatin regions has emerged as a widely used tool for identifying active regulatory elements in eukaryotes. However, existing approaches, limited by reliance on DNA fragmentation and short-read sequencing, cannot provide information about large-scale chromatin states or reveal coordination between the states of distal regulatory elements. We have developed a method for profiling the accessibility of individual chromatin fibers, a single-molecule long-read accessible chromatin mapping sequencing assay (SMAC-seq), enabling the simultaneous, high-resolution, single-molecule assessment of chromatin states at multikilobase length scales. Our strategy is based on combining the preferential methylation of open chromatin regions by DNA methyltransferases with low sequence specificity, in this case EcoGII, an N
6
-methyladenosine (m
6
A) methyltransferase, and the ability of nanopore sequencing to directly read DNA modifications. We demonstrate that aggregate SMAC-seq signals match bulk-level accessibility measurements, observe single-molecule nucleosome and transcription factor protection footprints, and quantify the correlation between chromatin states of distal genomic elements.
SMAC-seq combines long-read sequencing with open chromatin methylation by DNA methyltransferases to enable mapping of nucleosome position and chromatin accessibility.
Publisher
Nature Publishing Group US,Nature Publishing Group
