Massively Parallel Reporter Assay for pluripotency factors in mESCs

A massively parallel reporter assay, MPRA, was conducted in mouse embryonic stem cells (mESC). Synthetic cis-regulatory elements comprised of binding sites for pluripotency transcription factors and genomic sequences with comparable binding sites configurations were used in the assay. Transcripts of dsRed were amplified via PCR from the end of the transcript to sequence 3' UTR barcodes. MPRA from RW.4 mESCs transfected with either Synthetic (SYN) element library or a Genomic ('GEN' or 'g') WT/MUT sequence library. Each unique sequence or element of interest is tagged with 8 unique 9bp barcodes (provided in the description field in each sample record). Each library also includes a basal control (basal promoter only) tagged with 112 unique 9bp barcodes. The original DNA plasmids that were used for transfections were sequenced to use for normalizating plasmid copy number for final expresson calculations. Tested SYN element and gWT/gMUT sequence names are included in processed data files. These data are part of SRA submission SUB4287756 (SRP153192) and BioProject PRJNA480686.