An NT sgRNA, a KRAS-targeted sgRNA, and the 16 potential SOS2-targeted sgRNAs were each cloned into pLentiCRISPRv2, as previously described (33). sgRNA sequences are given in table S1. Lentiviruses were produced as described above. Forty-eight hours after infection, cells were selected for 4 days in puromycin at 6 μg/ml and then shifted into puromycin at 3 μg/ml for an additional 6 days. Ten days after selection, cells were analyzed for KRAS and SOS2 expression and plated for signaling and transformation assays.

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