CDK5RAP3 regulates hepatocyte differentiation in human

CDK5RAP3, an UFL1 adaptor for UFMylation, is crucial for liver development in mouse. However, the function of CDK5RAP3 in human hepatocyte differentiation remains unclear. The stepwise generation of hepatocyte-like cells derived from human embryonic cells simulates human hepatocyte differentiation in vitro. Here, the human embryonic cells were conducted to differentiate into hepatocytes and ScRNA-Seq analysis was performed to elucidate the molecular mechanism of CDK5RAP3 in human liver development. We found CDK5RAP3 was important for human hepatocytes differentiation. The CDK5RAP3 deficiency upregulated ER stress in hepatic-endoderm stage, and impaired ribosome homeostasis in mature hepatocytes. Overall design: Firstly, CDK5RAP3 gene is knockout in human embryonic cell (hESCs) line H9. Secondly, CDK5RAP3-null hESCs were conducted to differentiate into hepatocytes. The cells were collected on differentiated stage (10 and 25 days). Finally, ScRNA-Seq analysis was performed to elucidate the molecular mechanism of CDK5RAP3 in human liver development.