Fig. 5.

Genes encoding AAGAB, AP1 and AP2 are dispensable for cell survival or proliferation. To identify essential genes, we took advantage of a genome-wide HeLa mutant library generated using the GeCKO V2 CRISPR library (Gulbranson et al., 2017; Wang et al., 2019). The mutant cells were cultured continuously for four weeks before the abundance of gRNAs was analyzed by deep sequencing. Depletion of gRNAs was calculated by comparing the passage control population to the initial GeCKO V2 CRISPR DNA library. A CRISPR score was calculated for each gene based on log₁₀(fold-changes) in the abundance of its corresponding gRNAs using an established algorithm (Li et al., 2014). The 1391 genes with CRISPR scores below the horizontal cutoff line of −0.25 are predicted to be essential to cell survival or growth. Only the 14 genes encoding AAGAB, AP1 and AP2 are shown. A full list of genes ranked according to their CRISPR scores is shown in Table S5.