Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA.
Nat Biotechnol. 2019 Aug;37(8):916-924. doi: 10.1038/s41587-019-0147-6. Epub 2019 Jun 24.
Recent technical advancements have facilitated the mapping of epigenomes at single-cell resolution; however, the throughput and quality of these methods have limited their widespread adoption. Here we describe a high-quality (10 nuclear fragments per cell) droplet-microfluidics-based method for single-cell profiling of chromatin accessibility. We use this approach, named 'droplet single-cell assay for transposase-accessible chromatin using sequencing' (dscATAC-seq), to assay 46,653 cells for the unbiased discovery of cell types and regulatory elements in adult mouse brain. We further increase the throughput of this platform by combining it with combinatorial indexing (dsciATAC-seq), enabling single-cell studies at a massive scale. We demonstrate the utility of this approach by measuring chromatin accessibility across 136,463 resting and stimulated human bone marrow-derived cells to reveal changes in the cis- and trans-regulatory landscape across cell types and under stimulatory conditions at single-cell resolution. Altogether, we describe a total of 510,123 single-cell profiles, demonstrating the scalability and flexibility of this droplet-based platform.
最近的技术进步使得能够以单细胞分辨率绘制表观基因组图谱;然而,这些方法的通量和质量限制了它们的广泛采用。在这里,我们描述了一种基于高质量(每个细胞 10 个核片段)液滴微流控技术的单细胞染色质可及性分析方法。我们使用这种名为“通过测序进行转座酶可及染色质的液滴单细胞分析”(dscATAC-seq)的方法,对成年小鼠大脑中的 46653 个细胞进行无偏倚的细胞类型和调控元件的发现。我们通过将其与组合索引(dsciATAC-seq)相结合,进一步提高了该平台的通量,从而能够在大规模水平上进行单细胞研究。我们通过测量 136463 个静止和刺激的人骨髓来源细胞中的染色质可及性来证明该方法的实用性,从而在单细胞分辨率下揭示了细胞类型和刺激条件下顺式和反式调控景观的变化。总之,我们描述了总共 510123 个单细胞图谱,证明了这种基于液滴的平台的可扩展性和灵活性。