SummaryPeroxisome proliferator-activated receptor gamma (PPARγ) is a nuclear receptor central in the regulation of key cellular processes including cell metabolism, tissue differentiation, and regulation of the immune system. PPARγ is required for normal differentiation of the urothelium and is thought to be an essential driver of the luminal subtype of bladder cancer. However, the molecular components that regulatePPARGgene expression in bladder cancer remain unclear. Here, we developed an endogenousPPARGreporter system in luminal bladder cancer cells and performed genome-wide CRISPR knockout screening to identifybona fideregulators ofPPARGgene expression. Functional validation of the dataset confirmed GATA3, SPT6, and the cohesin complex components SMC1A, and RAD21, as permissive upstream positive regulators ofPPARGgene expression in luminal bladder cancer. In summary, this work provides a resource and biological insights to aid our understanding ofPPARGregulation in bladder cancer.