Fig. 2

Multiple CRISPR screens in HeLa cells for cholesterol regulatory genes. A The cell growth analysis of HeLa cells after introducing indicated sgRNAs via lentiviral infection for 7 days. Cells were counted with a hemacytometer. Mean ± SD with n = 3. Ordinary one-way ANOVA with Tukey’s test, ***p < 0.001. B The relative cell growth determined by cell counting for HeLa cells introducing indicated sgRNAs and treated with indicated doses of lovastatin. Mean ± SD with n = 3. Ordinary one-way ANOVA with Dunnett’s test, **p < 0.01, ***p < 0.001. C The workflow of genome-scale synthetic lethal CRISPR screens (Screen 2) to identify negative GIs with HMGCR using its inhibitor lovastatin in HeLa cells. D The scatter plot showing the β score of each gene and the correlation of both CRISPR screens (vehicle and lovastatin) in HeLa cells. The genes in blue box are preferential targets as the synthetic lethal or negative GI hits. E The top selected functional terms enriched among synthetic lethal hits of the CRISPR screens (Screen 2) as determined by the GO and KEGG analysis. F The workflow of genome-scale synthetic lethal CRISPR screens (Screen 3) to identify negative GIs with LDLR in LDLR KO single clone and AAVS1 KO control HeLa cells. G The rank-ordered list of each gene in the CRISPR screens (Screen 3) according to the strength of synthetic lethality measured by differential tRRA scores between LDLR KO single clone and AAVS1 KO control HeLa cells. The top interesting gene hits are highlighted. H Venn diagram showing the overlap of synthetic lethal or negative GI hits between the two independent LDLR KO clones in the CRISPR screens (Screen 3). I Venn diagram showing the overlap of synthetic lethal or negative GI hits between the three CRISPR screens. J Heatmap showing the relative strength of GIs of top selected hits to HMGCR or LDLR across different CRISPR screens