A genome wide CRISPR/Cas9 screening to identifiy potential embryonic diapause and diapause-like cancer cell survival genes

Diapause-like cancer cells represent a fuel for tumor relapse and strategies that leads to their elimination could be helpful in the context of minimal residual disease and tumor relapse. Here, through a genome-wide CRISPR/Cas9 screen performed in mouse embryonic stem cells undergoing diapause, we identify different genes potentially involved in the survival of diapause-like cancer cells. Further validation experiments in cancer cell lines led to the identification of the pathway NRF2-KEAP1, the 1C metabolism and the DNA methyltransferase DNMT1 as pathways involved in the survival of diapause-like cancer cells. Mouse embryonic stem cells (mESC) were induced to diapause via INK128 treatment (100nM). 5 days after the diapause induction, Cas9 was activated in diapause cells by the addition of doxycycline to the media for 3 days. Doxycyline was also adminstered to proliferating mESC. Screen was conducted in biological triplicate.