Discovery of NANOG enhancers and their essential roles in self-renewal and differentiation in human embryonic stem cells

Human embryonic stem cells (hESCs) are an important system to study development and regeneration. NANOG is a core regulator in hESCs, but it is transcriptionally regulated is not fully understood. In this study, we report the discovery of two NANOG enhancers from a CRISPR interference enhancer screen in hESCs. These enhancers are essential to hESCs, and deletion of a single copy of either enhancers significantly reduced NANOG expression. In addition, hESCs with heterozygous deletion of either enhancers had compromised self-renewal and increased tendency to differentiate. Interestingly, these two NANOG enhancers are involved in a tandem duplication event, but the duplicated counterparts do not regulate NANOG. This work expands our knowledge of functional enhancers in hESCs, and highlights the sensitivity of the hESC state to the dosage of a core regulator. Bulk RNA-seq of WT and NANOG enhancer knockout on embryonic stem cell and neuroectoderm *************************************************************** The table below lists GEO accessions reused/reanalyzed for this study. ***************************************************************