Figure 1.

Schematic representation of the workflow for the genome-wide CRISPR/Cas9 screen. HEK293FT cells were transfected with lentiviral CRISPR-Cas9 plasmid library (Brunello library with genome wide coverage and 4 sgRNAs per gene) for lentivirus production. A375 cells were then transduced with lentiviruses generated from the Brunello plasmid library. Cells were selected for successful transduction using puromycin selection for 7 days. After that, cells were either treated with 2 µM vemurafenib or DMSO. Samples were collected at days 7 and 14. gDNA was prepared from the cells and the relative sgRNA abundance was determined by PCR-mediated amplification of sgRNA sequences from gDNA followed by NGS.