Semi-automated IT-scATAC-seq profiles cell-specific chromatin accessibility in differentiation and peripheral blood populations

Wei Jin (), Jingchun Ma, Li Rong, Shengshuo Huang, Tuo Li, Guoxiang Jin and Zhongjun Zhou ()
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Wei Jin: Guangdong Medical University
Jingchun Ma: The University of Hong Kong
Li Rong: The University of Hong Kong
Shengshuo Huang: The University of Hong Kong
Tuo Li: Changzheng Hospital
Guoxiang Jin: Southern Medical University
Zhongjun Zhou: Guangdong Medical University

Nature Communications, 2025, vol. 16, issue 1, 1-13

Abstract: Abstract Single-cell ATAC-seq (scATAC-seq) enables high-resolution mapping of chromatin accessibility but is often limited by throughput, cost, and equipment requirements. Here, we present indexed Tn5 tagmentation-based scATAC-seq (IT-scATAC-seq), a semi-automated, cost-effective, and scalable approach that leverages indexed Tn5 transposomes and a three-round barcoding strategy. This workflow prepares libraries for up to 10,000 cells in a single day, reduces the per-cell cost to approximately $0.01, and maintains high data quality. Comprehensive benchmarking demonstrates that IT-scATAC-seq achieves robust library complexity, high signal specificity, and improved cost-efficiency compared to existing methods. We apply IT-scATAC-seq to mouse embryonic stem cells, capturing chromatin remodelling during early differentiation, and to human peripheral blood mononuclear cells, resolving cell-type–specific regulatory programs. Here, we show that IT-scATAC-seq provides a robust and efficient approach for high-resolution single-cell epigenomic investigations, balancing scalability, data quality, and accessibility.

Date: 2025
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DOI: 10.1038/s41467-025-57931-2

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