An abundance of binary promoter switches in lineage-determining transcription factors indicates a digital component of cell fate determination

An examination of transcription factor genes revealed an abundance of convergent and divergent sense/antisense promoter pairs, suggesting that competing promoters may control cell fate determination. We used 5' single cell RNAseq (5' scRNAseq) to examine divergent and convergent transcription in CD34+ hematopoietic progenitors, freshly purified innate lymphocytes from tonisl, and peripheral blood NK cells Overall design: Peripheral blood NK cells were purified using the RosetteSep Human NK Cell Enrichment Cocktail (STEMCELL Technologies). CD34+ hematopoietic progenitor cells were enriched using the Indirect Human CD34 MicroBead Kit (Miltenyi Biotec). Following cell sorting, cells were stained for flow cytometry and CD34+ cells were further purified via sorting to >99.9% purity on an FACSAria II cell sorter (BD Biosciences).Purified CD34+ progenitors were cultured for 2 weeks with OP9 stroma, FLT3-ligand, KIT-ligand, and IL3. Human innate lymphoid cells were enriched from fresh tonsil tissue and blood samples using a custom human NK cell enrichment RosetteSep reagent containing a mixture of bivalent antibodies against glycophorin A and CD3, CD4, CD19, CD20, CD36, CD66b, and CD123.