Abstract
Calling cards technology using self-reporting transposons enables the identification of DNA-protein interactions through RNA sequencing. By introducing a DNA barcode into the calling card itself, we have drastically reduced the cost and labor requirements of calling card experiments in bulk populations of cells. An additional barcode incorporation during reverse transcription enables simultaneous transcriptome measurement in a facile and affordable protocol. We demonstrate that barcoded self-reporting transposons recover in vitro binding sites for four basic helix-loop-helix transcription factors with important roles in cell fate specification: ASCL1, MYOD1, NEUROD2, and NGN1. Further, simultaneous calling cards and transcriptional profiling during transcription factor overexpression identified both binding sites and gene expression changes for two of these factors. RNA-based identification of transcription factor binding sites and gene expression through barcoded self-reporting transposon calling cards and transcriptomes is a novel and powerful method to infer gene regulatory networks in a population of cells.
Competing Interest Statement
The authors have declared no competing interest.