Schematic of Tuba-seq ^Ultra vector design and tumor initiation. U6-integrated diverse BC (U6^BC) contains a 21-nucleotide region at the 3′ end of the bovine U6 promoter immediately downstream of the TATA box. The sgRNA-Pool/Cre consists of barcoded lenti-sgRNA/Cre vectors that contain three sgRNAs targeting each gene of interest, safe cutting “inert” sgRNAs, and sgRNAs targeting an essential gene. KT;H11^LSL-Cas9 are transduced with the lenti- U6^BCsgRNA-Pool/Cre. 14–15 wk after tumor initiation, we extract DNA from bulk tumor-bearing lungs and used Tuba-seq^Ultra to quantify the impact of targeting each regulator on tumor growth, and tumor initiation for each tumor of each genotype.