In-organoid single-cell CRISPR screening reveals determinants of hepatocyte differentiation and maturation. Mus musculus

Harnessing differentiated and mature hepatocytes for basic research and regenerative medicine demands understanding its underlying principles. Despite tremendous efforts that have been made to generate functional hepatocytes from stem cells, the immature phenotype necessitates deeper investigations of the genetic circuit of differentiation and maturation. Single-cell CRISPR (scCRISPR) screens could link genetic perturbations with parallel transcriptomic readout in single cells, providing a powerful method to delineate multiple genes' contributions to a complex biological process. Here, we have developed a method named Organoid-based Single-cell CRISPR screening Analyzed with Regulons (OSCAR) to functionally assess candidate genes regulating hepatocyte differentiation and maturation.