Zhang Qiang, Ma Sai, Liu Zhengzhi, Zhu Bohan, Zhou Zirui, Li Gaoshan, Meana J Javier, González-Maeso Javier, Lu Chang
Department of Chemical Engineering, Virginia Tech, Blacksburg, VA 24061, USA.
Department of Biomedical Engineering and Mechanics, Virginia Tech, Blacksburg, VA 24061, USA.
bioRxiv. 2023 May 28:2023.05.26.542421. doi: 10.1101/2023.05.26.542421.
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 all 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 d. We applied the technology to profile mixed cell lines, mouse and human brain tissues to reveal cell type heterogeneity. Drop-BS will pave the way for single-cell methylomic studies requiring examination of a large cell population.
全基因组DNA甲基化图谱，即DNA甲基化组，是整体表观基因组格局的关键组成部分，可调节基因活性和细胞命运。单细胞DNA甲基化组学研究为基于甲基化组特征检测细胞亚群和进行细胞亚群分析提供了前所未有的分辨率。然而，现有的单细胞甲基化组学技术均基于使用试管或孔板，而这些平台在处理大量单细胞时不易扩展。在此，我们展示了一种基于微滴的微流控技术Drop-BS，用于构建用于DNA甲基化组分析的单细胞亚硫酸氢盐测序文库。Drop-BS利用微滴微流控提供的超高通量，在2天内制备多达10000个单细胞的亚硫酸氢盐测序文库。我们将该技术应用于分析混合细胞系、小鼠和人类脑组织，以揭示细胞类型的异质性。Drop-BS将为需要检测大量细胞群体的单细胞甲基化组学研究铺平道路。
