Programming and reprogramming neural cell types using synthetic transcription factors

摘要

An efficient method to produce large numbers of desirable human cell types in the laboratory is a major goal of stem cell research. Current approaches have focused on a strategy of recapitulating the events of normal embryogenesis in culture, by treating cells in stepwise protocols with cocktails of growth factors, pharmaceutical agents or forced expression of master regulatory transcription factors. Recently a new opportunity has arisen to exploit synthetic transcription factors (sTFs) to program and reprogram cell fate. sTFs are built by tethering various transcriptional regulatory effector domains (e.g. VP16, KRAB) to TALE or Cas9 engineered DNA binding domains. Here we report progress with the production and testing of sTFs designed to activate expression of Sox10, Olig2 and Nkx6-2 - reprogramming factors known to drive conversion of mouse embryonic fibroblasts (MEFs) to oligodendrocyte progenitor cells (OPCs). We explore whether such sTFs can operate in multiplex to increase the efficacy of reprogramming compared to cDNA overexpression. Ultimately we wish to assess if highly efficient genome-wide transcriptional resetting is possible through delivery of pools of sTFs to simultaneous activate or repress large cohorts of target genes.