Jul 1, 2016

Simultaneous measurement of chromatin accessibility, DNA methylation, and nucleosome phasing in single cells

BioRxiv : the Preprint Server for Biology
Sebastian Pott

Abstract

Gaining insights into the regulatory mechanisms that underlie the transcriptional variation observed between individual cells necessitates the development of methods that measure chromatin organization in single cells. Here I adapted Nucleosome Occupancy and Methylome-sequencing (NOMe-seq) to measure chromatin accessibility and endogenous DNA methylation in single cells (scNOMe-seq). scNOMe-seq recovered characteristic accessibility and DNA methylation patterns at DNase hypersensitive sites (DHSs). An advantage of scNOMe-seq is that sequencing reads are sampled independently of the accessibility measurement. scNOMe-seq therefore controlled for fragment loss, which enabled direct estimation of the fraction of accessible DHSs within individual cells. In addition, scNOMe-seq provided high resolution of chromatin accessibility within individual loci which was exploited to detect footprints of CTCF binding events and to estimate the average nucleosome phasing distances in single cells. scNOMe-seq is therefore well-suited to characterize the chromatin organization of single cells in heterogeneous cellular mixtures.

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Mentioned in this Paper

di-n-hexyl sulfosuccinate
DNA Methylation [PE]
Nucleosomes
Patterns
Deoxyribonuclease I
CTCF
Transcription, Genetic
DNA Methylation
Nucleic Acid Sequencing
Sequence Determinations

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