Essential transcription factors for human neuron differentiation

This project aims to identify key regulatory effectors downstream of NEUROG1/2 that are responsible for driving neural differentiation in human pluripotent stem cells. We aim to profile transcription factor (TF) expression and chromatin accessibility during the initial stages of NEUROG1/2-induced neuronal differentiation and construct directed graph networks of regulator and target TFs. We seek to gain a better understanding of the molecular mechanisms involved in the development of cortical-like neurons and to identify essential transcription factors that play a critical role in the differentiation process.We used previously established NYGCe001 hESCs with doxycycline-inducible overexpression of Neurogenin 2 and Neurogenin 1 (NEUROG2/1) that we differentiated into induced neurons (iNs).We next sought to connect regulator TFs to their target TFs by integrating RNA-sequencing and ATAC-sequencing data at various time points during differentiation. We specifically, we performed ATAC-seq on wild type hESCs at stem cell (ES) stage and 5 time points upon doxycycline induction. For RNA-seq, we sequenced libraries at ES and 5 time points upon doxycycline. In addition, we generated two NYGCe001 hESCs ZBTB18 -/- knockout lines and sequenced at ES and day 4 after doxycycline induction.