sgRNA-seq
For the pooled genome-wide CRISPR screen, 3E+8 PC-9 FR and H1975 FR cells were transfected with the pooled lentiviral TKOv3 library at a multiplicity of infection of 0.3-0.4. After 5 days of puromycin selection, the surviving cells were divided into 2 parts, each of which was treated with or without Furmonertinib. PCR was performed on genomic DNA to construct the sequencing library. Each library was sequenced at 2327 million reads to achieve 30 average coverage over the CRISPR library, and the data were analyzed by MAGeCK (version 0.5.9.5) and MAGeCK-VISPR (version 0.5.6). This whole-genome CRISPR/Cas9-sgRNA pooled library was purchased from Addgene (Cambridge, Massachusetts, USA).