CRISPR Screen: The Interferon ? Pathway Enhances Pluripotency and X-Chromosome Reactivation in iPSC Reprogramming

To investigate the genes and pathways involved in X-chromosome reactivation and pluripotency acquisition, we generated a whole-genome CRISPR knockout screen during female NPC reprogramming into iPSCs. We analyzed the gRNA abundance in NPCs and three different reprogramming populations: non-pluripotent, early pluripotent and late pluripotent. Pair-wise comparisons of gRNA frequencies in different populations were done to identify genes and pathways involved in cell survival in reprogramming conditions, colony formation, early pluripotency acquisition and late pluripotency/X-chromosome reactivation. Overall design: DNA sequencing of CRISPR-gRNA libraries in NPCs and cells undergoing reprogramming with different pluripotency and X-chromosome status.