GPR75 Knockout 786-O Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout population derived from the 786-O human renal cell carcinoma line. This product features a heterogeneous pool of cells carrying targeted gene disruptions in GPR75, an orphan G protein-coupled receptor. The polyclonal format maintains genetic diversity, avoiding single-cell cloning bottlenecks, and offers a representative loss-of-function model for studying GPR75 signaling and its metabolic roles. It is suitable for examining receptor function in the context of clear cell carcinoma.
The parental 786-O cell line originates from a primary clear cell adenocarcinoma of the kidney and serves as an established model of clear cell renal cell carcinoma (ccRCC). These adherent epithelial cells retain VHL deficiency, leading to constitutive HIF pathway activation and a dependence on glycolytic metabolism. The ccRCC background provides a relevant system for dissecting how GPCR signaling intersects with oncogenic metabolic reprogramming, as GPCR expression and cAMP dynamics are frequently altered in renal cancers.
GPR75 encodes an orphan GPCR that couples predominantly to G??s heterotrimeric G proteins, stimulating adenylyl cyclase to elevate intracellular cAMP. This rise in cAMP activates PKA, which phosphorylates targets such as CREB, and additionally engages PI3K/AKT and MAPK/ERK cascades. ??-arrestins interact with GPR75, potentially mediating receptor desensitization or G protein-independent signaling. Although its endogenous ligands remain unknown, GPR75 signaling converges on pathways that regulate cellular energy homeostasis and gene expression.
In 786-O cells, GPR75 knockout enables dissection of GPCR-driven modulation of metabolic vulnerabilities inherent to ccRCC. The receptor??s downstream effectors??including PKA, AKT, and ERK1/2??intersect with metabolic checkpoints, and loss of GPR75 may perturb cAMP-dependent phosphorylation events. This model is valuable for exploring crosstalk between GPCR pathways and the VHL-HIF axis central to ccRCC pathogenesis.
Applications include cAMP accumulation assays and phospho-signaling analysis to quantify second messenger responses and kinase activation downstream of GPR75. RT-qPCR and western blotting can profile changes in PI3K/AKT and MAPK pathway gene and protein expression. Co-immunoprecipitation identifies GPR75-associated protein complexes, including heterotrimeric G proteins and ??-arrestins. Functional studies employ metabolic flux analysis and cell proliferation assays to assess metabolic reprogramming and growth phenotypes. The model is also applicable to drug target validation for obesity and metabolic syndrome, where GPR75 has been linked to insulin resistance. For technical inquiries, please contact Ascent Research.
