Fig. 1: Pooled CRISPR knockout screen of IL-1β regulators in human monocytic cells U937.

a Treatment of PMA-differentiated U937-LC3-Cas9+ cells with increasing doses of LPS induces expression and secretion of IL-1β. Addition of Nigericin augments the secretion of IL-1β. Values are represented as an average of three independent replicates -/+ standard deviation (SD). b Intracellular IL-1β level upon induction with LPS compared to vehicle treatment. Values are represented as an average of three independent replicates -/+ SD. Statistical significance was determined using two-sided, unpaired Student’s t test (*P < 0.05). c Overview of the genome-wide CRISPR screening strategy using the Brunello sgRNA library. The diagram was created with BioRender.com. Genes that decrease (d) or increase (e) IL-1β levels when knocked out in U937-LC3-Cas9+ cells. For each gene the x axis shows the enrichment of sgRNAs in log₂ enrichment values, and the y axis shows statistical significance of enrichment as -log₁₀P value. Statistically significant genes were identified by aggregating differential sgRNAs ranked consistently higher at gene level using the robust rank aggregation (RRA) method. Pathways and classes of known and unknown genes that modulate IL-1β levels identified in IL-1β^low (f) and IL-1β^high (g) populations. LPS lipopolysaccharide, sgRNA single guide RNA, NGS next-generation sequencing.