Double knockout CRISPR screen for cancer resistance to T cell cytotoxicity

To systematically interrogate genetic interactions that mediate cancer cells' response to T cell killing, we designed an asymmetric CRISPR/Cas9 library targeting the matched combinations between significantly mutated cancer genes and immune resistance genes with available compounds targeting involved pathways. We performed a combinatorial double knockout screen on gene pairs and identified those where joint loss-of-function renders altered cellular response to T cell cytotoxicity.