Self-Reporting Transposons Enable Simultaneous Readout of Gene Expression and Transcription Factor Binding in Single Cells

Cellular heterogeneity confounds in situ assays of transcription factor (TF) binding. Single-cell RNA sequencing (scRNA-seq) deconvolves cell types from gene expression, but no technology links cell identity to TF binding sites (TFBS) in those cell types. We present self-reporting transposons (SRTs) and use them in single-cell calling cards (scCC), a novel assay for simultaneously measuring gene expression and mapping TFBS in single cells. The genomic locations of SRTs are recovered from mRNA, and SRTs deposited by exogenous, TF-transposase fusions can be used to map TFBS. We then present scCC, which map SRTs from scRNA-seq libraries, simultaneously identifying cell types and TFBS in those same cells. We benchmark multiple TFs with this technique. Next, we use scCC to discover BRD4-mediated cell-state transitions in K562 cells. Finally, we map BRD4 binding sites in the mouse cortex at single-cell resolution, establishing a new method for studying TF biology in situ. [Display omitted] •Novel genetic element can be mapped from mRNA and directed by DNA-binding proteins•Combined gene expression and transcription factor binding data from single cells•Mapped multiple transcription factors in several cell lines and the mouse cortex•Discovered bromodomain-dependent cell-state transitions in leukemic cells Moudgil et al. present a single-cell method for simultaneously capturing gene expression and transcription factor binding site data from the same cells, first in cell lines and then in the mouse brain.