Fig. 1: CRISPR-Cas9 functional genomics screen identifies NUDT1 as a selective MYC-driven metabolic dependency.

a Schematic depicting the pooled CRISPR-based screen. SHEP MYCN-ER-Cas9 cells were transduced with a metabolism-focused library of lentiviral sgRNAs targeting 2745 metabolic genes. At population doubling 0 (PD 0), cells were cultured with or without 4-OHT treatment (50 nM) for 14 population doublings (PD 14). Relative abundance of sgRNAs in PD 14 was compared to those in PD 0. b Gene scores in untreated versus 4-OHT treated SHEP MYCN-ER cells. The gene score means the median log₂ fold change of all sgRNAs abundance targeting each metabolic gene during the culture period. The analysis was performed utilizing MAGeCK (version 0.5.9.5), which involved the calculation of p-values. c Abundance changes in the primary pooled screen of the individual NUDT1 sgRNAs with or without 4-OHT treatment. d The biochemical reaction catalyzed by NUDT1. NUDT1 hydrolyzes the representative oxidized triphosphates 8-oxo-dGTP into 8-oxo-dGMP, which is unable to incorporate into DNA. e Cell death analysis of SHEP MYCN-ER cells treated with 4-OHT (200 nM) for 24 h and infected with the NUDT1 shRNA or control (shNC) lentivirus particles. f Cell death analysis of P493 cells treated with Tetracycline (Tet, 100 ng/ml) for 24 h and infected with the NUDT1 shRNA or control (shNC) lentivirus particles. g Immunoblots of MYCN and MYC in various tumor cells, with β-Actin as a loading control. h Cell death analysis of various tumor cells infected with the NUDT1 sgRNA or control (sgNC) lentivirus particles. i Cell death analysis of NUDT1-depleted Kelly cells expressing NUDT1 wild-type (WT) or the catalytically dead mutant (E56A). e, f, h, i data are shown as averages of technical triplicates; e–i these experiments were independently repeated three times with similar results. Statistical significance was determined by two-way ANOVA (e, f, i). Source data are provided as a Source Data file.