Fig. 2: Identification of ATP7A as a vulnerability for KRAS-addicted CRC cells.
From: Copper bioavailability is a KRAS-specific vulnerability in colorectal cancer
a Experimental design for identification of KRAS-specific vulnerabilities by in vitro CRISPR/Cas9 screening. b Graph depicting the fold-change distributions of gRNAs targeting essential (solid lines) and nonessential (dashed lines) genes at day 7 after infection of Control and KRAS cells with KRAS-library. c Waterfall graph showing differential essentiality scores between Control and KRAS cells, depicting genes coding for cell-surface proteins significantly altered by KRASG12V (red diamonds) and reference genes coding for proteins known to be essential (green circles) or non-essential (blue squares). d Bar graphs illustrating the Log2 FC of the eight gRNAs targeting Atp7a, the non-essential gene Gpr101 and essential gene Myc at day 7 in Control (blue) and KRAS (red) cells. e Bar chart indicating the relative cell viability of Control and KRAS cells after ATP7A knockdown. IB for ATP7A showing the efficient knockdown by RNAi (right). Data represent n = 3 independent experiments with five replicates, and n = 3 independent experiments (right). ***P = 0.0001; ****P < 0.0001. f As in e CRC cells were analyzed for cell viability upon ATP7A knockdown. Graph showing the efficient knockdown of ATP7A as measured by qPCR (right). Data represent n = 3 independent experiments with seven replicates, and two technical replicates (right). ****P < 0.0001 (CACO-2 versus HCT116, shATP7A-22); ***P = 0.0008 (CACO-2 versus SW620, shATP7A-22); **P = 0.0027 (CACO-2 versus HCT116, shATP7A-26); *P < 0.0001 (CACO-2 versus SW620, shATP7A-26). g Experimental design for the identification of KRAS-specific vulnerabilities in vivo using CRISPR/Cas9. h Waterfall graph showing BF scores for genes coding for cell-surface proteins significantly regulated by KRASG12V (red diamonds), or proteins that are essential (green circles) or non-essential (blue squares). For panels e, f center values and error bars represent mean ± SEM, and significance was determined using two-way ANOVA with post-hoc Bonferroni’s multiple-comparison analysis.
