title: Profiling DNA methylome landscapes of mammalian cells with single-cell reduced-representation bisulfite sequencing.
abstract: The heterogeneity of DNA methylation within a population of cells necessitates DNA methylome profiling at single-cell resolution. Recently, we developed a single-cell reduced-representation bisulfite sequencing (scRRBS) technique in which we modified the original RRBS method by integrating all the experimental steps before PCR amplification into a single-tube reaction. These modifications enable scRRBS to provide digitized methylation information on ∼1 million CpG sites within an individual diploid mouse or human cell at single-base resolution. Compared with the single-cell bisulfite sequencing (scBS) technique, scRRBS covers fewer CpG sites, but it provides better coverage for CpG islands (CGIs), which are likely to be the most informative elements for DNA methylation. The entire procedure takes ∼3 weeks, and it requires strong molecular biology skills.
authors.affiliation: Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing, China.
authors.forename: Hongshan
authors.initials: H
authors.lastname: Guo
authors.affiliation: 1] Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing, China. [2] Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.affiliation: Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Guo
authors.affiliation: Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing, China.
authors.forename: Xianlong
authors.initials: X
authors.lastname: Li
authors.affiliation: 1] Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing, China. [2] Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China.
authors.forename: Xinglong
authors.initials: X
authors.lastname: Wu
authors.affiliation: Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing, China.
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Fan
authors.affiliation: 1] Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing, China. [2] Ministry of Education Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking University, Beijing, China. [3] Center for Molecular and Translational Medicine, Peking University Health Science Center, Beijing, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Wen
authors.affiliation: 1] Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing, China. [2] Ministry of Education Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking University, Beijing, China. [3] Center for Molecular and Translational Medicine, Peking University Health Science Center, Beijing, China.
authors.forename: Fuchou
authors.initials: F
authors.lastname: Tang
mesh_terms: D000818:Animals; D018899:CpG Islands; D019175:DNA Methylation; D005260:Female; D023281:Genomics / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D008297:Male; D008322:Mammals / Q000235:genetics*; D051379:Mice; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods; D011786:Quality Control; D059010:Single-Cell Analysis / Q000379:methods*; D013447:Sulfites
chemical_list: D013447:Sulfites; C042345:hydrogen sulfite
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