A CRISPR/Cas9-based kinome screen identifies ErbB signaling as a new regulator in the regulation of naïve pluripotency and totipotency in humans

Regulation of totipotency and naïve pluripotency in humans is crucial for early human embryo development. However, the mechanisms of naïve pluripotency and totipotency regulation in humans, especially the signaling pathways that are involved in these processes, remains largely unknown. We performed a CRISPR/Cas9-based kinome knockout screen to analyze the effect of disrupting 763 human kinases in regulating human naïve pluripotency. Further validation using small molecules revealed that inhibition of ErbB family promoted the transition of hEPSCs to human naïve pluripotent stem cells. More importantly, chemical inhibition of ErbB family also promoted induction of totipotent signatures in human pluripotent cells under different culturing conditions. A total of 9 samples were analyzed by bulk RNA sequencing, two biological replicates of each sample, which included primed, extended and naïve pluripotent stem cells with or without the treatment of Afatinib or Erlotinib for 7 days under respective culture conditions.