Fig. 1.

Epigenetic Domain-specific Knock Out Library (EpiDoKOL) screen identified common essentiality genes in T98G and U373 cells. A Composition of EpiDoKOL including target molecules, their functions and related epigenetic modifications. B Library generation procedure. Figure created with BioRender.com. C Cas9 activity assay of U373 and T98G cells. Microscopic images of T98G and U373 cells transduced with indicated sgRNAs 18 days post-transduction. D Flow cytometric analysis of U373 and T98G cells transduced with indicated sgRNAs (g-NT1, g-NT2: non-targeting sgRNAs; g-T1 and g-T2: sgRNAs targeting GFP), up to 12 days post-transduction. E Schematic of EpiDoKOL screening procedure. Figure created with BioRender.com. F sgRNA density plots from cells transduced with pLentiCRISPRv2 plasmid containing EpiDoKOL. Cell pellets collected before puromycin selection. G Log2 counts of sgRNAs at initial and final time points in T98G and U373 cells. H Waterfall plots for Log2fold changes of genes after screening T98G cells with EpiDoKOL for a month. Domain and gene-based analysis revealed overlapping essentiality hits. I Waterfall plots for Log2fold changes of genes after screening U373 cells with EpiDoKOL for a month. Domain and gene-based analysis revealed overlapping essentiality hits. Mageck p values were calculated with negative binomial model fitting. Other p values were determined by two-tailed Student’s t-test *P < 0.05, **P < 0.01, ***P < 0.001