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. 2022 Jan 27;21(4):594–606. doi: 10.1158/1535-7163.MCT-21-0532

Figure 1.

Figure 1. Silencing of apoptotic pathway genes confers resistance to agonists of the death receptor pathway. Volcano plots of genes enriched or depleted for gRNA when rTRAIL-treated cells were compared with DMSO vehicle control cells following transduction with a genome-wide CRISPR/Cas9 library in MSTO-211H mesothelioma (A) and PC-9 lung adenocarcinoma (B) cells. Each dot represents gene-wise scores for MAGeCK gene level analysis with genes of interest highlighted. The dotted line indicates an FDR of 0.1. x-axis, log2 fold change of mean gRNA reads per genes comparing treatment to DMSO replicates. y-axis, −log10 FDR for genes based upon their negative (pink and red dots) and positive (blue dots) fold change and FDR calculated by the MAGeCK algorithm. C, Confirmation of deletion of target apoptotic genes following transfection with synthetic crRNA in MSTO-211H, NCI-H28, and H2804 cancer cells. D, Six-day viability assay in MSTO-211H cells following deletion of specific apoptotic genes and treatment with either rTRAIL or MEDI3039. y-axis: viability effect relative to control cells. E, Volcano plots as in A and B illustrating results of SAM CRISPR activatory screen in PC-9 cells treated with rTRAIL. F, MSTO-211H isogenic cell lines were screened versus the parental cell line with a concentration range of 60 compounds and viability measured at day 6. The AUC values for each isogenic cell line and the matched parental Cas9 line were subtracted to calculate a ΔAUC value, with high (positive) values indicating increased resistance to that compound in the isogenic lines, and low (negative) values increased sensitivity. x-axis: name of compounds screened. y-axis: ΔAUC values.

Silencing of apoptotic pathway genes confers resistance to agonists of the death receptor pathway. Volcano plots of genes enriched or depleted for gRNA when rTRAIL-treated cells were compared with DMSO vehicle control cells following transduction with a genome-wide CRISPR/Cas9 library in MSTO-211H mesothelioma (A) and PC-9 lung adenocarcinoma (B) cells. Each dot represents gene-wise scores for MAGeCK gene level analysis with genes of interest highlighted. The dotted line indicates an FDR of 0.1. x-axis, log2 fold change of mean gRNA reads per genes comparing treatment to DMSO replicates. y-axis, −log10 FDR for genes based upon their negative (pink and red dots) and positive (blue dots) fold change and FDR calculated by the MAGeCK algorithm. C, Confirmation of deletion of target apoptotic genes following transfection with synthetic crRNA in MSTO-211H, NCI-H28, and H2804 cancer cells. D, Six-day viability assay in MSTO-211H cells following deletion of specific apoptotic genes and treatment with either rTRAIL or MEDI3039. y-axis: viability effect relative to control cells. E, Volcano plots as in A and B illustrating results of SAM CRISPR activatory screen in PC-9 cells treated with rTRAIL. F, MSTO-211H isogenic cell lines were screened versus the parental cell line with a concentration range of 60 compounds and viability measured at day 6. The AUC values for each isogenic cell line and the matched parental Cas9 line were subtracted to calculate a ΔAUC value, with high (positive) values indicating increased resistance to that compound in the isogenic lines, and low (negative) values increased sensitivity. x-axis: name of compounds screened. y-axis: ΔAUC values.