CRISPR screening in human primordial germ cell-like cells specification

Although molecular studies on the human reproduction are arising, mechanisms underlying human primordial germ cell (PGC) development have remained largely unknown. Here, we conducted custom CRISPR screening to identify a subset of key genes for acquisition and maintenance of PGC fate in humans, combined with in vitro system of hPGC-like cells (hPGCLCs). Among crucial candidates by screen analysis, we characterize AKT coactivator, TCL1A, as a progressive role in hPGCLC development. A mutant TCL1A downregulated gene expression related to translational control, followed by reduced global protein synthesis and proliferation consequently via depressing AKT-mTOR signaling pathway. Our study provides novel regulators critical to hPGCLC development and demonstrate the importance of translational control in human reproduction. hiPSCs, iMeLCs, hPGCLCs, from wild-type and TCL1A knockout, were used in this study.