kinome-CRISPR screening with Cal27 cells using INK128

To investigate the mechanism of mTOR inhibitors in head and neck squamous cell carcinoma (HNSCC), a CRISPR screen was performed. The sgRNA library used was the Human Kinome CRISPR pooled library (Brunello). Briefly, Cal27-Cas9 cells were infected with Human Kinome CRISPR pooled library, and were treated with 2 different groups; vehicle treated or INK128 (10nM) treated group until total population doubling reached 20. the barcode 31 was PCR-recovered from genomic samples, and samples where sequenced to calculate 32 abundance of the different sgRNA probes. PCR of sgRNA for Illumina sequencing protocol was 33 obtained from the Broad Institute.