Fig. 2: Secondary targeted CRISPR knock-out screen in HUDEP-2 cells.

A Overview of the secondary (validation) CRISPR knock-out screen, in which HUDEP-2 cells were transduced with the custom targeted lentiviral library, cultured for 9 days in maintenance media, and subsequently differentiated for 12 days. Cells were collected both prior to differentiation and live CD49d^low CD233⁺ cells were sorted at day 12 of differentiation. sgRNA abundance was compared among the secondary CRISPR library and the latter two cell populations. Volcano plots displaying MAGeCK gene-level enrichment scores on the y-axis and average log₂-fold change of sgRNA abundance of each gene on the x-axis for (B) day 0 versus library analysis and (C) day 12 versus day 0 analysis. Improved False Discovery Rates (FDR) in the secondary versus genome-scale screens for several characterized (D) erythroid essential genes and (E) genes required for erythroid differentiation. Improved FDRs in the targeted compared to genome-scale screen for virtually all genes included in both screens, for both (F) day 0 versus library analysis and (G) day 12 versus day 0 analysis. Source data are provided in Source Data file.