CoBATCH for High-Throughput Single-Cell Epigenomic Profiling
CoBATCH for High-Throughput Single-Cell Epigenomic Profiling
复制标题
用于高通量单细胞表观基因组分析的 CoBATCH
DOI：
10.1016/j.molcel.2019.07.015
复制
发表时间：
2019-10-03
期刊：
影响因子：
16
通讯作者：
He, Aibin
中科院分区：
文献类型：
--
作者：
Wang, Qianhao;Xiong, Haiqing;He, Aibin
单细胞组蛋白修饰或染色质结合蛋白的全基因组图谱缺乏一种有效的、可推广的方法。在这里，我们开发了CoBATCH，组合条形码和靶向染色质释放，用于单细胞图谱的染色质结合蛋白在细胞培养和组织中的基因组分布。融合到Tn5转座酶的蛋白A通过针对基因组区域的特异性抗体而丰富，Tn5产生索引的染色质片段，准备用于文库准备和测序。重要的是，这一策略不仅能够在完整的组织中进行低投入的表观基因组图谱，而且还可以测量在自然条件和交叉连接条件下每个实验可扩展到多达数万个单细胞。CoBATCH以极低的本底产生类似于12,000个读取/细胞的结果。通过CoBATCH绘制10个胚胎小鼠器官的内皮细胞谱系图，可以有效地破译细胞群体的表观遗传异质性和顺式调控机制。因此，CoBATCH不需要专门的设备，可广泛应用于蛋白质-DNA相互作用的单细胞图谱分析，并可轻松部署。
An efficient, generalizable method for genome-wide mapping of single-cell histone modifications or chromatin- binding proteins is lacking. Here, we develop CoBATCH, combinatorial barcoding and targeted chromatin release, for single-cell profiling of genomic distribution of chromatin-binding proteins in cell culture and tissue. Protein A in fusion to Tn5 transposase is enriched through specific antibodies to genomic regions, and Tn5 generates indexed chromatin fragments ready for library preparation and sequencing. Importantly, this strategy enables not only low-input epigenomic profiling in intact tissues but also measures scalable up to tens of thousands of single cells per experiment under both native and cross-linked conditions. CoBATCH produces similar to 12,000 reads/cell with extremely low background. Mapping of endothelial cell lineages from ten embryonic mouse organs through CoBATCH allows for efficient deciphering of epigenetic heterogeneity of cell populations and cis-regulatory mechanisms. Thus, obviating specialized devices, CoBATCH is broadly applicable and easily deployable for single-cell profiling of protein-DNA interactions.