Integration of significant genes hit across 3 time points of etoposide exposures. (A) Venn diagram of significant genes identified by CRISPR screening at day 4 (purple), day 12 (green), and day 18 (orange), with 8 EtoSGs (blue) and 3 EtoRGs (red), were common at all time points. (B) Eleven EtoSGs/EtoRGs at all time points of etoposide exposure. (C) Eight EtoSGs/EtoRGs at day 4 and day 12 of etoposide exposure. (D) Three EtoSGs/EtoRGs at day 4 and day 18 of drug exposure. (E) Twenty-eight EtoSGs/EtoRGs at day 12 and day 18 of etoposide exposure. (F) Twenty-five significant genes with etoposide time dependence score derived from MAGeCK-MLE, where fitting time linearity of etoposide score with time points of day 0 (time 0), day 4 (time 1), day 12 (time 2), and day 18 (time 3), where high etoposide time dependence score indicating genes exhibit consistent etoposide response changes. All significant genes were identified at FDR < 0.1. (G) Spearman rank correlation between significant EtoRGs genes with only negative etoposide score (resistance score) and their corresponding essential score at each time point. The etoposide score was determined by comparing etoposide to vehicle control at each time point, whereas the essential score was determined by comparing vehicle control at each time point to day 0, with “ns” indicating P > .05.