A combinatorial indexing strategy for epigenomic profiling of plant single cells

Abstract

Understanding how cis-regulatory elements facilitate gene expression is a key question in biology. Recent advances in single-cell genomics have led to the discovery of cell-specific chromatin landscapes that underlie transcription programs. However, the high equipment and reagent costs of commercial systems limit their applications for many laboratories. In this study, we profiled the Arabidopsis root single-cell epigenome using a combinatorial index and dual PCR barcode strategy without the need of any specialized equipment. We generated chromatin accessibility profiles for 13,576 Arabidopsis thaliana root nuclei with an average of 12,784 unique Tn5 integrations per cell and 85% of the Tn5 insertions localizing to discrete accessible chromatin regions. Comparison with data generated from a commercial microfluidic platform revealed that our method is capable of unbiased identification of cell type-specific chromatin accessibility with improved throughput, quality, and efficiency. We anticipate that by removing cost, instrument, and other technical obstacles, this combinatorial indexing method will be a valuable tool for routine investigation of single-cell epigenomes and usher new insight into plant growth, development and their interactions with the environment.

Time-resolved single-cell and spatial gene regulatory atlas of plants under pathogen attack

“…A majority of links were within a short distance (<400bp; potential promoter regions), but many links were farther away from genes (potential enhancer regions) (Figures 2B-2C). Cluster marker genes (1,774 genes) had more linked ACRs in a moderately distal region (2.5 kb to 5 kb from TSS) compared with non-marker genes (Figure 2D), implying the role of CREs moderately distal from the TSS in cell type/state-specific gene regulation, which is consistent with the observation in a previous study (Tu et al, 2021). We also found that linked ACRs were slightly larger than non-linked ACRs (Figure S2A).…”