Wide-genome CRISPR screening with cetuximab using HNSCC cell line

Cal27-Cas9 cells were infected with Human Genome-wide CRISPR pooled library (Brunello) that was a gift from John Doench and David Root (73178-LV, Addgene), and were treated with 2 different groups; vehicle treated or cetuximab (200ug/mL) treated group. The treatment was continued until the population doubling reached 20. The genomic DNA was extracted using DNeasy Blood and Tissue Kit (69504, QIAGEN). The entirety of isolated genomic DNA was used for subsequent PCR to ensure capturing the full representation of the libraries. PCR products were purified using NucleoSpin Gel and PCR Clean-Up Kit (740609, Takara Bio) and sequenced on a NovaSeq6000 (Illumina).