SEC24A identified as an essential mediator of thapsigargin-induced cell death in a genome-wide CRISPR/Cas9 screen.

Endoplasmic reticulum (ER) stress from accumulated misfolded proteins in the ER can activate the unfolded protein response (UPR). The UPR acts either to restore proteostasis or to activate cell death pathways if the stress cannot be resolved. The key downstream effectors in these pathways have been studied extensively. However, in comparison, stressor-specific key mediators are not as well characterized. In this study, we sought to identify and compare the genes that are necessary for cell death induced by three classic pharmacological ER stressors with different mechanisms of action: thapsigargin, tunicamycin, and brefeldin A. We conducted genome-wide CRISPR/Cas9-based loss-of-function screens against these agents in HAP1 cells, which are a near-haploid cell line. Our screens confirmed that MFSD2A and ARF4, which were identified in previous screens, are necessary for tunicamycin- and brefeldin A-induced cytotoxicity, respectively. We identified a novel gene, SEC24A, as an essential gene for thapsigargin-induced cytotoxicity in HAP1 cells. Further experiments showed that the ability of SEC24A to facilitate ER stress-induced cell death is specific to thapsigargin and that SEC24A acts upstream of the UPR. These findings show that the genes required for ER stress-induced cell death are specific to the agent used to induce ER stress and that the resident ER cargo receptor protein SEC24A is an essential mediator of thapsigargin-induced UPR and cell death.