Fig. 1: Genome-wide CRISPR screen identified oxidative stress-related genetic factors.

A Diagram of genome-wide CRISPR screen. B Western blotting of Cas9 in stable Cas9-expressing HEK293 cells. C Genomic DNA PCR with the primers flanking the two gRNA target sites in the DMD gene detected a small deletion product in Cas9⁺ HEK293 cells transfected with the gRNA pair and in control HEK293 cells transfected with both Cas9 and gRNA pair, but not in control or Cas9⁺ cells without gRNA transfection. The scatter (D) and volcano (E) plots showing enrichment and depletion of specific gRNAs after H₂O₂ treatment. Control-pre cells are the Cas9⁺ HEK293 cells harboring the gRNA library before H₂O₂ treatment, while the control cells showing in the X-axis are the Cas9⁺ HEK293 cells harboring the gRNA library without H₂O₂, but otherwise similarly cultured as for the cells treated with H₂O₂. H₂O₂ sample represents Cas9⁺ HEK293 cells harboring the gRNA library with H₂O₂. F Gene ontology (GO) enrichment analysis of the genes with p < 0.05. Number of genes belonging to each category is indicated. BP biological process, MF molecular function. G Validation of the top six hits using the CCK8 viability assay. The OD450 ratio means the ratio of the OD450 value after H₂O₂ treatment for 6 days to the initial OD450 value on day 0. *p < 0.05, ****p < 0.0001 (one-way ANOVA).