The GPR75 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of the human CAL-27 oral squamous cell carcinoma line. This product offers a heterogeneous pool of cells with targeted disruption of the GPR75 gene, creating a loss-of-function model in an epithelial cancer background. The polyclonal format preserves diverse genetic modifications while eliminating GPR75-mediated signaling, making it suitable for robust functional studies of this orphan GPCR.
CAL-27 is an adherent epithelial cell line derived from a tongue squamous cell carcinoma, commonly used in head and neck cancer research. These cells maintain malignant epithelial traits and active intracellular signaling pathways that intersect with GPCR cascades. Hosting the GPR75 knockout in this line enables investigation of receptor function within the context of oncogenic signaling, metabolic regulation, and tumor biology, providing a clinically relevant platform for dissecting GPR75-dependent mechanisms.
GPR75 functions as a Gq/11-coupled receptor activated by 20-hydroxyeicosatetraenoic acid (20-HETE). Upon ligand binding, it stimulates phospholipase C ?? (PLC??), which generates inositol trisphosphate (IP3) and diacylglycerol (DAG), leading to intracellular calcium mobilization and activation of protein kinase C (PKC). This cascade promotes phosphorylation of extracellular signal-regulated kinase (ERK) and the transcription factor CREB, ultimately modulating gene expression programs associated with energy homeostasis. GPR75 also interacts with ??-arrestin and influences AMPK signaling, broadening its metabolic regulatory scope. Knockout cells lack this 20-HETE-dependent signaling axis.
In the CAL-27 squamous carcinoma background, disruption of GPR75 signaling reveals the crosstalk between metabolic sensing and cancer cell behavior. Since 20-HETE and downstream effectors??including PKC, ERK, and CREB??can modulate proliferation, migration, and survival, this model permits dissection of GPR75??s role in oncogenic processes. The loss of these signals in a tumor-derived epithelial line helps elucidate how orphan GPCRs may contribute to malignant phenotypes, such as altered glucose metabolism and invasive properties. Thus, the knockout model bridges metabolic signaling research and squamous carcinoma biology.
This polyclonal knockout pool supports diverse functional assays: calcium flux measurements for GPCR activity, phospho-ERK western blotting for MAPK pathway analysis, MTS assays for proliferation, and glucose uptake assays for metabolic profiling. RT-qPCR of downstream target genes enables transcriptional analysis, while Transwell migration and Annexin V apoptosis assays evaluate invasive and survival characteristics. Applications include metabolic disease modeling, anti-obesity drug screening, GPCR functional studies, hypertension research, and cancer cell signaling investigations. For additional information, contact Ascent Research.
