In this current study, a genome‐wide CRISPR‐Cas9‐based genetic screening with the Brunello CRISPR knockout library was carried out for the first time in cisplatin‐resistant NSCLC Calu1 cells that identified GPR89A, PUF60, NBAS and GRPEL1 as novel candidate genes. Here, PUF60 is linked to non-small cell lung carcinoma.