Fig. 6

Multiplexed, quantitative analysis of Kras mutant oncogenicity using AAV/Cas9-mediated somatic HDR and high-throughput sequencing of barcoded lung tumors. a Pipeline to quantitatively determine the number, size, and genotype of individual tumors directly from bulk lung samples by high-throughput sequencing of tumor barcodes. b–e Number of lung tumors harboring each mutant Kras allele normalized to its initial representation (mutant representation in the AAV plasmid library divided by WT representation in the AAV plasmid library) and relative to WT (mutant tumor # divided by WT tumor #). Variants present in significantly more tumors than WT (two-sided Fisher’s exact test; p < 0.05) are colored blue; darker blue indicates no significant difference from G12D (p > 0.05), lighter blue indicates significantly less tumors with the indicated variant than G12D (p < 0.05). Error bars are bootstrapped 95% confidence intervals. Bar plots were generated from pooled data from all mouse genotypes (N = 15) (b), or individually from LT;H11 ^LSL-Cas9 (N = 6) (c), PT;H11 ^LSL-Cas9 (N = 6) (d), or T;H11 ^LSL-Cas9 (N = 3) (e) mice. Note that different y axis scales are used in each plot