title: Single-cell methylome landscapes of mouse embryonic stem cells and early embryos analyzed using reduced representation bisulfite sequencing.
abstract: DNA methylation is crucial for a wide variety of biological processes, yet no technique suitable for the methylome analysis of DNA methylation at single-cell resolution is available. Here, we describe a methylome analysis technique that enables single-cell and single-base resolution DNA methylation analysis based on reduced representation bisulfite sequencing (scRRBS). The technique is highly sensitive and can detect the methylation status of up to 1.5 million CpG sites within the genome of an individual mouse embryonic stem cell (mESC). Moreover, we show that the technique can detect the methylation status of individual CpG sites in a haploid sperm cell in a digitized manner as either unmethylated or fully methylated. Furthermore, we show that the demethylation dynamics of maternal and paternal genomes after fertilization can be traced within the individual pronuclei of mouse zygotes. The demethylation process of the genic regions is faster than that of the intergenic regions in both male and female pronuclei. Our method paves the way for the exploration of the dynamic methylome landscapes of individual cells at single-base resolution during physiological processes such as embryonic development, or during pathological processes such as tumorigenesis.
authors.affiliation: Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing 100871, China;
authors.forename: Hongshan
authors.initials: H
authors.lastname: Guo
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.forename: Xinglong
authors.initials: X
authors.lastname: Wu
authors.forename: Xianlong
authors.initials: X
authors.lastname: Li
authors.forename: Lu
authors.initials: L
authors.lastname: Wen
authors.forename: Fuchou
authors.initials: F
authors.lastname: Tang
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000235:genetics*; D002478:Cells, Cultured; D018899:CpG Islands; D019175:DNA Methylation; D004622:Embryo, Mammalian / Q000378:metabolism*; D047108:Embryonic Development / Q000235:genetics; D053595:Embryonic Stem Cells / Q000378:metabolism*; D044127:Epigenesis, Genetic; D005260:Female; D005306:Fertilization; D016678:Genome; D006238:Haploidy; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D058977:Molecular Sequence Annotation; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods*; D059010:Single-Cell Analysis; D013094:Spermatozoa / Q000378:metabolism; D015053:Zygote / Q000378:metabolism