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. Author manuscript; available in PMC: 2016 Mar 12.
Published in final edited form as: Cell. 2015 Mar 5;160(6):1246–1260. doi: 10.1016/j.cell.2015.02.038

Figure 6.

Figure 6

(A) Schematic representation of the loss-of-function metastasis minipool screen. Briefly, Cas9-GFP KPD cells were transduced with either validation minipool (524 gene-targeting + 100 non-targeting sgRNAs) or control minipool (624 non-targeting sgRNAs). After puromycin selection, the cell pools are transplanted into Nu/Nu mice. After 6 weeks, validation minipool sgRNAs are sequenced from primary tumor and lung samples.

(B) Primary tumor growth curve of Nu/Nu mice transplanted with Cas9 vector + validation minipool cells (n = 5 mice) or Cas9 + control minipool cells (n = 5 mice).

(C) Percent of lung lobes with metastases after 6 weeks for the mice in (B). C = control minipool. V = validation minipool.

(D) Boxplot of the sgRNA normalized read counts for the plasmid library, cells before transplantation, primary tumor and lung metastases using the validation minipool.

(E) Cumulative probability distribution of library sgRNAs in the validation plasmid pool, cells before transplantation, early tumor and lung metastases. Distributions of primary tumor and lung metastases are averaged across 5 mice.

(See also: Figure S7)