CRISPR/Cas9 screening

The experimental group received oxaliplatin (20 mg/kg) every two days. After one month, oxaliplatin-treated and control tumors were harvested, and genomic DNA was extracted using a DNA extraction and purification kit (Qiagen, Hilden, Germany) as previously described 14. sgRNA inserts were amplified by polymerase chain reaction (PCR) using NEBNext High Fidelity PCR Master Mix (NEB, Ipswich, MA, USA). For each sample, 10 μg of gDNA was used as input for PCR amplification. PCR products were then used to construct paired-end libraries with the Paired-End DNA Sample Prep Kit (Illumina Inc., San Diego, CA, USA). Deep sequencing was performed on a NovaSeq 6000 platform (Illumina Inc.) at Genedenovo Biotechnology Co., Ltd. (Guangzhou, China).