The GPR75 Knockout AGS Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma epithelial cell line. This tool enables loss-of-function studies of GPR75, an orphan G protein-coupled receptor, in a gastric cancer context. The polyclonal format provides a heterogeneous pool of cells with targeted disruption of the GPR75 locus, facilitating robust and reproducible functional assays without the clonal selection bias associated with monoclonal lines.
AGS cells are a widely utilized model derived from a primary human gastric adenocarcinoma. They retain key characteristics of gastric epithelial cells and are commonly employed in gastric cancer research, including studies of tumorigenesis, cell signaling, and metabolic regulation. Their adherent epithelial morphology and well-characterized genetic background make them suitable for knockout and overexpression experiments, as well as for drug screening and mechanistic dissection of oncogenic pathways.
GPR75 encodes an orphan GPCR that transduces signals through heterotrimeric G proteins, primarily G??s and G??q/11. Upon activation by yet unidentified endogenous ligands or metabolic signals such as insulin, GPR75 stimulates adenylate cyclase via G??s, elevating intracellular cAMP levels and activating PKA, which then phosphorylates downstream targets including CREB. Alternatively, coupling to G??q/11 activates PLC??, generating IP3 and Ca2+ mobilization. These bifurcating pathways modulate key kinases such as Akt and ERK1/2, integrating insulin signaling and metabolic regulation. ??-arrestin-1 and ??-arrestin-2 also interact with GPR75, mediating receptor desensitization and scaffolding additional signaling cascades. In vivo genetic evidence links GPR75 to body weight homeostasis and insulin sensitivity, positioning it as a potential therapeutic target for obesity and type 2 diabetes.
Disruption of GPR75 expression in the AGS gastric adenocarcinoma background provides a physiologically relevant platform for investigating GPR75-mediated metabolic and oncogenic signaling in epithelial cells. Since gastric epithelial cells are exposed to nutrient and hormonal signals, this knockout model allows researchers to dissect how loss of GPR75 function affects cellular responses to insulin, glucose uptake, and downstream metabolic flux. Moreover, it enables the exploration of cross-talk between GPR75 signaling and gastric cancer progression, given the emerging links between metabolic dysregulation and tumorigenesis.
Researchers can employ this knockout cell population in a variety of assays to validate GPR75 as a drug target. Western blotting and RT-qPCR confirm knockout efficiency and assess pathway activation by monitoring phosphorylation of Akt (Ser473) and ERK1/2 (Thr202/Tyr204). Functional studies may include cAMP accumulation assays, calcium flux measurements, cell proliferation analyses, and glucose uptake assays. Metabolic flux analysis can further reveal alterations in glycolytic or oxidative metabolism. This product is particularly suited for high-content screening of ligands or compounds that modulate GPR75 activity and for mechanistic studies in metabolic disease modeling. For additional information, please contact Ascent Research.
