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CRISPR edited Knock-out (KO) cell lines are powerful tools to identify key biomarkers and study disease mechanisms, such as the regulation of pathways by the disruption of gene of interest. In this presentation, we highlight how our KO cell lines can be utilised to investigate signaling pathways. Here we present two case studies, firstly focussing on the TRAIL signaling pathway. By selecting a cell line lacking the TRAIL receptor (DR5), we demonstrate how downstream apoptotic signaling can be attenuated. Secondly, we established our A549 KO cells as physiologically relevant models to study interactions between lung cancer cells and T cells. Disruption of immune-based pathways by CRISPR gene editing is a powerful technique to elucidate the contribution of individual proteins in immune checkpoint signalling by cancer cells.