Single-cell mapping of regulatory DNA-protein interactions.

Publication Type Academic Article
Authors Chi W, Yoon S, Goksel E, Mekerishvili L, Pelt J, Lin Y, Prieto T, Zinno J, Ganesan S, Potenski C, Izzo F, Landau D, Raimondi I
Journal Cell
Volume 189
Issue 12
Pagination 3801-3816.e11
Date Published 06/04/2026
ISSN 1097-4172
Keywords Single-Cell Analysis, Transcription Factors, DNA-Binding Proteins, DNA
Abstract Gene expression is controlled by transcription factors (TFs), whose genome binding is shaped by chromatin accessibility and histone modifications, yet mapping these interactions, particularly those with weak affinity or a transient nature, in single cells remains technically challenging. To address this gap, we developed docking and deamination followed by sequencing (D&D-seq), a single-cell immuno-tethering technology for profiling DNA-protein interactions. D&D-seq couples an antibody-binding nanobody to a cytosine base editor, a combination that enables detection of weak or transient factor binding through targeted cytosine-to-uracil editing at protein-bound genomic sites. This approach is compatible with standard single-cell multi-omic workflows and therefore allows integrated analyses of gene regulation. Using assay for transposase-accessible chromatin using sequencing (ATAC-seq) and single-cell ATAC-seq (scATAC-seq), we assessed chromatin accessibility as a functional readout of TF activity, and by coupling D&D-seq with whole-genome sequencing, we captured CTCF binding in both active and inactive chromatin compartments.
DOI 10.1016/j.cell.2026.05.014
PubMed ID 42242226
PubMed Central ID PMC13366695
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