Fig. 3. Validation of EXOSC8^rs117135638 as a GEMINI vulnerability.

a Schematic of EXOSC8 SNP rs117135638 locus showing target sites for positive control, non-allele specific (NA) sgRNA and experimental, allele-specific (AS) sgRNA. Alleles appear in bold. b Crystal structure of EXOSC8 gene product, Rrp43⁸⁹ (gray) shows the amino acid encoded by rs117135638 (teal) lies on the surface of the Rrp43 protein near the interface with exosome complex subunit Mtr3 (orange). c Disruption of EXOSC8 in patient-derived EXOSC8 resistant (EXOSC8^R) or EXOSC8 sensitive (EXOSC8^S) cells expressing EXOSC8 non-allele specific (NA) positive control sgRNA or allele-specific (AS) experimental sgRNA. Unaltered alleles (black), alleles with in-frame insertions or deletions (gray), and alleles with frameshift alterations (yellow) were assessed by deep sequencing of EXOSC8 four days post-infection with sgRNA. d Immunoblot of EXOSC8 protein levels in indicated patient-derived and isogenic cell lines expressing LacZ, EXOSC8 NA, or EXOSC8 AS sgRNA (n = 2 technical replicates of 1 biological sample). e Representative growth curves of indicated patient-derived and isogenic cell lines expressing LacZ (black), EXOSC8 NA (red), or EXOSC8 AS (blue) sgRNA, as measured by CellTiter-Glo luminescence, relative to day of assay plating. n = 5 technical replicates. Data are presented as mean values ± s.d. See Supplementary Fig. 4 for additional biological replicates. f Immunoblot of EXOSC8 protein levels in indicated isogenic cell lines expressing LacZ, EXOSC8 NA, or EXOSC8 AS sgRNA (n = 2 technical replicates of 1 biological sample). g Representative growth curves of indicated isogenic cell lines expressing LacZ (black), EXOSC8 NA (red), or EXOSC8 AS (blue) sgRNA, as measured by CellTiter-Glo luminescence, relative to day of assay plating. n = 5 technical replicates. Data are presented as mean values ± s.d. See Supplementary Fig. 4 for additional biological replicates. Source data for Fig. 3c–g are provided as a Source Data file.