Genome-scale CRISPR Screen Reveals Essential Biological Process Determining Human Lineage Specification

Human pluripotent stem cells (hPSCs) have an unlimited capacity to self-renewal and generate any somatic cell types in the human body, offer a promising cell resource for disease modeling and transplantation. However, the systematic mechanism leading the conversion of hPSCs into lineage specification, which is the earliest cellular event during human embryonic development, is elusive. Here, combining a lineage-specific reporter with inducible CRISPR/Cas9 system, we conducted a genome-scale screening in hESCs for overall drawing the track of cell fate decision. We defined eight functional modules containing embryonic development, mRNA processing, metabolism process, epigenetic regulation, cell cycle and others, that participate in full to the lineage commitment. We performed genome-wide CRISPR/iCa9 loss-of-function (LOF) screen in hPSCs with lineage reporter.