Simultaneous profiling of DNA methylation and chromatin architecture in mixed populations and in single cells

Dynamic DNA methylation and three-dimensional chromatin architecture compose a major portion of a cells epigenome and play an essential role in tissue specific gene expression programs. Currently, DNA methylation and chromatin organization are generally profiled in separate assays. Here, we report Methyl-HiC, a method combining in situ Hi-C and whole genome bisulfite sequencing (WGBS) to simultaneously capture chromosome conformation and DNA methylome in a single assay. Methyl-HiC analysis of mouse embryonic stem cells reveals coordinated DNA methylation between distant yet spatially proximal genomic regions. Extension of Methyl-HiC to single cells further enables delineation of the heterogeneity of both chromosomal conformation and DNA methylation in a mixed cell population, and uncovers increased dynamics of chromatin contacts and decreased stochasticity in DNA methylation in genomic regions that replicate early during cell cycle.