In vitro and in vivo epigenome-wide CRISPR screens in mouse KrasG12D/P53-/- (KP) lung adenocarcinoma model

To systematically study the functions of epigenetic genes in controlling tumor progression and regulating anti-tumor immunity, we performed a series of in vitro and in vivo epigenome-wide CRISPR screens in mouse KP lung adenocarcinoma model. We constructed an epigenetic-focused sgRNA library, established KP-Cas9-library pools, and then performed in vitro and in vivo CRISPR screens. For in vitro screens, we compared the cell pools harvested at week 4 with the cells pools harvested at week 2 to check the functions of epigenetic genes in tumor cell proliferation. For in vivo screens, we compared the IgG treated tumors in Rag1-/- mice or WT mice with the input cell pool to check the functions of epigenetic genes in tumor growth; we compared the IgG treated tumors in WT mice with the IgG treated tumors in Rag1-/- mice to check the functions of epigenetic genes in regulating anti-tumor immunity; we compared the anti-PD-1 treated tumors in WT mice with IgG treated tumors in WT mice to check the functions of epigenetic genes in modulating sensitivity to anti-PD-1 treatment. Overall design: Examination of sgRNA abundance in library, cultured cell pools, input cell pool and tumors with different treatments.