Fig. 13

A Leveraging CRISPR/Cas9 to create drug-resistant cancer cell lines for use as advanced drug screening tools. Here, subfigure (i) illustrates the structure and working process of the CRISPR/hCas9 system. Subfigure (ii) depicts immunofluorescence analysis of MED12 protein levels in MED12KO cells using different stains. Cells were stained with Alexa 488-phalloidin, rabbit anti-MED12 antibody/rhodamine red-labeled goat anti-rabbit IgG, and DAPI. Subfigure (iii) shows western blot analysis confirming MED12 protein levels. Subfigure (iv) compares the response of BRAF inhibitor-resistant clones to various inhibitor combinations [Colors indicate different drug efficiencies: no significant inhibitory effect (white, cell coverage ≥ 80%), mild inhibitory effect (yellow, 60% ≤ cell coverage < 80%), moderate effect (orange, 30% ≤ cell coverage < 60%), and strong inhibitory effect (red, cell coverage < 30%)]. Adapted with permission from [236] (Copyright Elsevier, 2018). B A schematic overview of a novel strategy to prioritize targets in multiple cancer types by incorporating CRISPR–Cas9 gene fitness effects, genomic biomarkers and target tractability for drug development. Adapted with permission from [237] (Copyright Springer Nature, 2019)