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Systematic comparison of estimates of transcription factor activity by ATAC-seq and multiplexed reporter assays

van Steensel, B.; Trauernicht, M.; Franceschini-Santos, V.; Filipovska, T.

2026-01-05 genomics
10.64898/2026.01.05.697632 bioRxiv
Show abstract

Transcription factors (TFs) are central to gene regulation and play critical roles in development, cellular homeostasis and disease. The ability to accurately measure TF activity is essential to understanding how TFs respond to signals and regulate target genes. In one commonly used approach, activities of TFs are computationally inferred from genome-wide chromatin accessibility data (ATAC-seq). However, it has remained unclear how well these inferences reflect actual regulatory activity of TFs. An alternative approach employs a collection of synthetic reporters that are designed to each probe the regulatory activity of a single TF. In this study, we systematically compared TF activities as inferred by ATAC-seq with those measured by multiplexed reporters, across diverse perturbations known to alter specific TF activities. We observed considerable overlap between the two methods, but also notable discrepancies. Our findings suggest that reporter assays and chromatin-based inference capture distinct aspects of TF function: reporter assays are more sensitive to signal-responsive TFs, while ATAC-seq better detects chromatin-modifying TFs.

Published in Molecular Systems Biology (predicted rank #30) · training set

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