In Vivo CRISPR Screening for metabolic regulators of immunotherapy. Mus musculus

To systematically study the functions of metabolic genes in regulating anti-tumor immunity, we performed a series of in vivo metabolic-wide CRISPR screens in orthotopic lung tumor mice models. We constructed a metabolic-focused sgRNA library, established LLC-Cas9-library pools, and then performed in vivo CRISPR screens. We compared the PBS treated tumors in Rag1-/- mice or C57BL/6 mice with the input cell pool to check the functions of metabolic genes in tumor growth; we compared the PBS treated tumors in C57BL/6 mice with the PBS treated tumors in Rag1-/- mice to check the functions of metabolic genes in regulating anti-tumor immunity; we compared the anti-PD1 treated tumors in C57BL/6 mice with PBS treated tumors in C57BL/6 mice to check the functions of metabolic genes in modulating sensitivity to anti-PD1 treatment.