Skip to main content
. 2023 Oct 30;5(1):47–65. doi: 10.1038/s43018-023-00653-5

Fig. 3. Identification of key mediators of imetelstat efficacy using genome-wide CRISPR/Cas9 editing.

Fig. 3

Brunello CRISPR/Cas9 positive enrichment screen in NB4 cells. a, gRNA enrichment analysis using STARS and RIGER gene-ranking algorithms in n = 3 independent imetelstat-treated biological replicates. Red circles indicate significantly enriched targets (STARS false discovery rate (FDR) < 0.15 and RIGER score >2.0). b, Cytoscape visualization of the ingenuity pathway analysis (IPA)-derived interaction network connecting the identified significantly enriched gRNA targets. cf, Competition assays of imetelstat- (red) versus vehicle control (PBS; black)-treated Cas9-expressing NB4 (c), MV411 (d), KO52 (e) and TF1 (f) cultures transduced with n = 2 independent sgRNAs targeting FADS2 (top), n = 4 independent sgRNAs targeting ACSL4 (middle) and n = 2 controls (empty vector and gRNA targeting CD33). Three technical replicates per condition from two independent experiments were pooled. Asterisks (*) denote statistically significant comparisons based on distinct 95% CI on mCherry chimerism AUC between imetelstat and PBS-treated cultures. 95% CI (lower limit, upper limit): NB4 FADS2 PBS (444.4, 456.7) versus imetelstat (771.9, 795.9); ACSL4 PBS (320.1, 344.4) versus imetelstat (428.3, 459.6); editing controls PBS (186.0, 197.8) versus imetelstat (185.4, 203.4) (c). MV411 FADS2 PBS (1,253, 1,302) versus imetelstat (1,421, 1,525); ACSL4 PBS (847.8, 1,160) versus imetelstat (1,262, 1,564); editing controls PBS (896.2, 1,030) versus imetelstat (943.8, 1,073) (d). KO52 FADS2 PBS (812.3, 907.0) versus imetelstat (949.8, 1,023); ACSL4 PBS (648.9, 743.3) versus imetelstat (1,020, 1,095); editing controls PBS (635.3, 727.6) versus imetelstat (654.9, 760) (e). TF1 FADS2 PBS (1,280, 1,325) versus imetelstat (1,585, 1,721); ACSL4 PBS (1,381, 1,644) versus imetelstat (2,486, 2,788); editing controls PBS (905.9, 982.8) versus imetelstat (888.2, 993.3) (f).

Source data