DNA甲基化
表观遗传学
单细胞分析
计算生物学
甲基化
微流控
亚硫酸氢盐测序
CpG站点
甲基化DNA免疫沉淀
细胞
化学
计算机科学
DNA
生物
遗传学
纳米技术
基因
基因表达
材料科学
作者
Xi Zeng,Xiaoxu Guo,Shaowei Jiang,Xiaoping Yang,Zhixing Zhong,Siyu Liu,Zhi Zhu,Jia Song,Chaoyong Yang
标识
DOI:10.1021/acs.analchem.3c02484
摘要
Single-cell DNA methylation sequencing is highly effective for identifying cell subpopulations and constructing epigenetic regulatory networks. Existing methylome analyses require extensive starting materials and are costly, complex, and susceptible to contamination, thereby impeding the development of single-cell methylome technology. In this work, we report digital microfluidics-based single-cell reduced representation bisulfite sequencing (digital-scRRBS), the first microfluidics-based single-cell methylome library construction platform, which is an automatic, effective, reproducible, and reagent-efficient technique to dissect the single-cell methylome. Using our digital microfluidic chip, we isolated single cells in 15 s and successfully constructed single-cell methylation sequencing libraries with a unique genome mapping rate of up to 53.6%, covering up to 2.26 million CpG sites. Digital-scRRBS demonstrates a high capacity for distinguishing cell identity and tracking DNA methylation during drug administration. Digital-scRRBS expands the applicability of single-cell methylation methods as a versatile tool for epigenetic analysis of rare cells and populations with high levels of heterogeneity.
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