Figure 7.

(A) Pie charts of the most abundant sgRNAs in the primary tumor and the whole lung of two representative mice transplanted with validation minipool transduced Cas9-GFP KPD cells. The area for each sgRNA corresponds to the fraction of total reads from the tissue (primary tumor or lung metastases) for the sgRNA. All sgRNAs with ≥ 2% of total reads are plotted individually.

(B) Scatterplot of normalized sgRNA read counts in primary tumor and lung metastases for all sgRNAs in the validation minipool for each mouse (different color dots indicate sgRNAs from different mice). log₂ n.r., log₂ normalized reads.

(C) log₂ ratio of sgRNA abundance in the lung metastases over the primary tumor (Metastasis-Primary Ratio, or MPR) plotted against the abundance in the lung metastases (n = 5 mice per sgRNA). Green dots are the 100 control sgRNAs. Dots with black outlines are non-control sgRNAs that target genes or miRs. Red dots indicate non-control sgRNAs for which more than one sgRNA targeting the same gene/miR is enriched in the lung metastases over the primary tumor (i.e. log₂(MPR) > 0 ) and are labeled with the gene/miR targeted. The lung-primary ratio is calculated for individual mice and these quantities are averaged across mice.

(D) Number of genes with 0 to 10 significantly enriched validation minipool sgRNAs in lung metastases. For genes/miRs with 2 or more enriched sgRNAs, genes/miRs are categorized by how many sgRNAs targeting that gene/miRs are enriched as indicated in the colored bubbles adjacent to each bar.

(E) Schematic illustration of tumor growth and metastasis in the library-transduced NSCLC transplant model. The initially diverse set of loss-of-function mutations in the subcutaneously transplanted pool is selected over time for mutations that promote growth of the primary tumor. A subset of these mutants also dominate lung metastases.

(See also: Figure S7)