Abstract: The recent derivation of human trophoblast stem cells (hTSCs) offers a scalable in vitro model system for studying human placental development. However, the molecular mechanisms underlying hTSC differentiation into extravillous trophoblasts (EVT) remain unclear. In this study, we initially induced hTSC differentiation into EVT in vitro and collected RNA samples at day 0, 2, 4, and 6 for high-throughput sequencing to identify key genes involved in the differentiation process. We then employed a genome-wide CRISPR-Cas9 knockout screening to systematically narrow down the critical genes in hTSCs, selecting the top five percent for further functional validation. Currently, we are performing lineage-directed EVT differentiation on both wild-type and several gene knockout hTSCs . Overall, our research will contribute a valuable resource for understanding the molecular regulation of human placental development and associated diseases.
