CRISPR screen of genes encoding protein interactors of MCB-613 in parental and EGFR inhibitor resistant EGFR-mutant NSCLC cell lines

CRISPR screen of genes encoding protein interactors of MCB-613 in PC9, GR4, and WZR12 cells Overall design: PC9, GR4, and WZR12 cells were infected with a CRISPR library targeting genes encoding protein interactors of MCB-613 and carried forward in triplicate for 14 population doublings at 1000X coverage. Genomic DNA was extracted using the QIAamp Blood Maxi Kit (Qiagen). Extracted DNA was amplified using a two-step protocol involving staggered barcode sequences for pooled sequencing. Sequencing was performed on an Illumina NextSeq 500 with 75bp paired-end sequencing and analysis was performed using the Models-based Analysis of Genome-wide CRISPR/Cas9 Knockout (MAGeCK) software analysis pipeline under default conditions.