The GPR180 Knouckout SK-HEP-1 Polyclonal Cells product comprises a heterogeneous population of SK-HEP-1 cells engineered via CRISPR/Cas9-mediated disruption of the GPR180 gene. This polyclonal knockout model enables loss-of-function studies of the orphan G protein-coupled receptor GPR180 within a hepatic endothelial-like cellular background. The pooled cell population preserves the genetic and phenotypic diversity inherent to polyclonal editing, providing a robust tool for investigating receptor function without clonal selection bias.
The SK-HEP-1 parental line originates from the ascitic fluid of a male patient with liver adenocarcinoma and displays characteristics of hepatic sinusoidal endothelial cells. These cells serve as an established in vitro model for the liver sinusoidal endothelium, which is central to hepatic filtration, nutrient exchange, and immune surveillance. The endothelial-like phenotype of SK-HEP-1 cells includes expression of endothelial markers and the capacity to form capillary-like tubes, making them suitable for studies of vascular biology in a liver-specific context.
GPR180 encodes an orphan G protein-coupled receptor that is implicated in lipid metabolism and vascular development. Though its endogenous ligands remain unidentified, the receptor is predicted to respond to lipid-derived molecules and metabolic signals. Upon activation, GPR180 couples to heterotrimeric G?? proteins, engaging downstream effectors that include adenylyl cyclase, cAMP, and protein kinase A (PKA), as well as the MAPK/ERK cascade. ??-arrestins may also be recruited to the receptor, contributing to signal termination and G protein-independent signaling. Thus, GPR180 is positioned at the interface of metabolic sensing and intracellular signaling pathways that regulate cellular responses to lipid cues.
Disruption of GPR180 in the SK-HEP-1 hepatic endothelial-like background enables dissection of the receptor??s role in liver sinusoidal endothelial cell function. Because hepatic sinusoidal endothelium is a critical interface for lipid handling and metabolic regulation, the knockout model can be used to investigate how GPR180 influences endothelial barrier properties, lipid uptake, and angiogenic responses such as tube formation. The polyclonal knockout format avoids clonal artifacts and allows observation of population-level phenotypic changes, which is particularly relevant for studying heterogeneous endothelial behaviors.
Researchers can employ this polyclonal knockout product in a variety of assays to characterize GPR180 signaling and function. Western blotting and RT-qPCR enable confirmation of GPR180 knockout and assessment of downstream target expression changes, while Sanger sequencing verifies CRISPR-induced disruptions at the genomic level. Functional studies may include cAMP and calcium flux assays to interrogate G protein-dependent signaling, as well as lipid uptake assays and tube formation experiments to evaluate metabolic and angiogenic capabilities. These applications support GPCR deorphanization efforts, investigation of hepatic endothelial biology, and validation of GPR180 as a therapeutic target for dyslipidemia and metabolic syndrome. For further information, please contact Ascent Research.
