Multiplex indexing approach for the detection of DNase I hypersensitive sites in single cells

作者：Weiwu Gao, Wai Lim Ku, Lixia Pan, Jonathan Perrie, Tingting Zhao, Gangqing Hu, Yuzhang Wu, Jun Zhu, Bing Ni, Keji Zhao · 发表于：Nucleic Acids Research · 年份：2021 · DOI：10.1093/nar/gkab102 · 被引用次数：19 · 研究领域：Single-cell and spatial transcriptomics、Epigenetics and DNA Methylation、Genomics and Chromatin Dynamics

Single cell chromatin accessibility assays reveal epigenomic variability at cis-regulatory elements among individual cells. We previously developed a single-cell DNase-seq assay (scDNase-seq) to profile accessible chromatin in a limited number of single cells. Here, we report a novel indexing strategy to resolve single-cell DNase hypersensitivity profiles based on bulk cell analysis. This new technique, termed indexing single-cell DNase sequencing (iscDNase-seq), employs the activities of terminal DNA transferase (TdT) and T4 DNA ligase to add unique cell barcodes to DNase-digested chromatin ends. By a three-layer indexing strategy, it allows profiling genome-wide DHSs for >15 000 single-cells in a single experiment. Application of iscDNase-seq to human white blood cells accurately revealed specific cell types and inferred regulatory transcription factors (TF) specific to each cell type. We found that iscDNase-seq detected DHSs with specific properties related to gene expression and conservation missed by scATAC-seq for the same cell type. Also, we found that the cell-to-cell variation in accessibility computed using iscDNase-seq data is significantly correlated with the cell-to-cell variation in gene expression. Importantly, this correlation is significantly higher than that between scATAC-seq and scRNA-seq, suggesting that iscDNase-seq data can better predict the cellular heterogeneity in gene expression compared to scATAC-seq. Thus, iscDNase-seq is an attractive alternativ...