Department of Chemical Engineering, Virginia Tech, Blacksburg, VA, 24061, USA.
Department of Biomedical Engineering and Mechanics, Virginia Tech, Blacksburg, VA, 24061, USA.
Nat Commun. 2023 Aug 3;14(1):4672. doi: 10.1038/s41467-023-40411-w.
The genome-wide DNA methylation profile, or DNA methylome, is a critical component of the overall epigenomic landscape that modulates gene activities and cell fate. Single-cell DNA methylomic studies offer unprecedented resolution for detecting and profiling cell subsets based on methylomic features. However, existing single-cell methylomic technologies are based on use of tubes or well plates and these platforms are not easily scalable for handling a large number of single cells. Here we demonstrate a droplet-based microfluidic technology, Drop-BS, to construct single-cell bisulfite sequencing libraries for DNA methylome profiling. Drop-BS takes advantage of the ultrahigh throughput offered by droplet microfluidics to prepare bisulfite sequencing libraries of up to 10,000 single cells within 2 days. We apply the technology to profile mixed cell lines, mouse and human brain tissues to reveal cell type heterogeneity. Drop-BS offers a promising solution for single-cell methylomic studies requiring examination of a large cell population.
全基因组 DNA 甲基化图谱（或 DNA 甲基组）是整体表观基因组景观的关键组成部分，可调节基因活性和细胞命运。单细胞 DNA 甲基组学研究提供了前所未有的分辨率，可根据甲基组特征检测和分析细胞亚群。然而，现有的单细胞甲基组学技术基于使用管或孔板，这些平台难以扩展以处理大量的单细胞。在这里，我们展示了一种基于液滴的微流控技术 Drop-BS，用于构建单细胞亚硫酸氢盐测序文库以进行 DNA 甲基组分析。Drop-BS 利用液滴微流控技术提供的超高通量，在 2 天内可制备多达 10000 个单细胞的亚硫酸氢盐测序文库。我们将该技术应用于混合细胞系、小鼠和人脑组织的分析，以揭示细胞类型异质性。Drop-BS 为需要检查大量细胞群体的单细胞甲基组学研究提供了有前途的解决方案。
