A multiplexed in vivo gene knockout approach to identify cancer drivers in small cell lung cancer [SCLC Tuba-seq]

We generated a model of lentivirus-initiated mouse small cell lung cancer (SCLC) combined with CRISPR/Cas9 knockout and barcoding. We analyzed tumor suppressive functions of 39 gene candidates using these in vivo CRISPR screening approaches and found Tsc1 to be one of the top tumor suppressor genes in SCLC. mRNA profiles of tumors initiated from Rb/p53/Rbl2-mutant (RPR2) mouse model using adenoviral and lentiviral vectors, as well as sequencing of sgRNA identifier-barcode (sgID-BC) amplicons from genomic DNA extracted from whole mouse lung or laser capture microdissected sections.