The GPR75 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9?edited polyclonal cell population derived from the human HCT 116 colorectal carcinoma cell line in which the GPR75 gene has been disrupted. This heterogeneous knockout pool serves as a ready?to?use loss?of?function model for investigating the orphan G protein?coupled receptor GPR75 in metabolic regulation and GPCR signaling. By eliminating GPR75 expression across a mixed population, this product enables robust functional comparisons with wild?type HCT 116 cells without requiring single?cell cloning.
The host HCT 116 cell line originates from a male patient with colorectal adenocarcinoma and harbors a KRAS G13D mutation while maintaining wild?type TP53, making it a widely employed epithelial cancer model. These adherent cells exhibit rapid proliferation, facilitating diverse downstream assays. The colorectal carcinoma background provides a relevant context for examining GPR75 function, as GPCRs can influence both tumor cell metabolism and signaling crosstalk. Although GPR75 is primarily linked to energy homeostasis, its expression in HCT 116 cells permits exploration of potential metabolic regulatory roles in epithelial lineages.
GPR75 is an orphan GPCR that couples to the G??s subunit to stimulate adenylyl cyclase, leading to increased intracellular cAMP levels. This triggers downstream PKA activation and subsequent CREB phosphorylation, constituting a canonical cAMP?mediated signaling cascade. Metabolic factors such as insulin and glucose reportedly regulate GPR75 expression, though its endogenous ligand remains unknown. Receptor activity is further modulated by interactions with beta?arrestin and other GPCR?associated proteins. Through this pathway, GPR75 may transcriptionally influence genes involved in energy expenditure and insulin secretion, as suggested by human loss?of?function variants associated with lower adiposity.
In the HCT 116 model, CRISPR?mediated disruption of GPR75 allows researchers to dissect its contribution to GPCR?cAMP?PKA?CREB signaling within an epithelial cancer cell environment. Although HCT 116 cells are derived from colorectal carcinoma, they retain core metabolic machinery, making them suitable for probing GPR75?dependent metabolic responses. This system can reveal how an orphan receptor modulates cAMP dynamics and downstream transcriptional programs, shedding light on its possible roles in nutrient sensing or energy balance. The polyclonal nature ensures that genetic heterogeneity is maintained, mimicking a more physiologically diverse cell population compared to monoclonal knockouts.
The GPR75 Knockout HCT 116 Polyclonal Cells are designed for advanced biomedical research applications, including mechanistic dissection of GPCR signal transduction and drug target validation in metabolic disorders. The product supports quantitative assays such as cAMP accumulation measurements, phospho?CREB Western blotting, and CRE?luciferase reporter gene activation. Additionally, it enables functional metabolic studies like glucose uptake assays, oil red O staining for lipid accumulation, and RT?qPCR of metabolic genes. These cells are a valuable tool for functional genomics in obesity, type 2 diabetes, and metabolic syndrome research. Their polyclonal nature ensures broad representation of knockout events, increasing experimental robustness. For further information or technical support, please contact Ascent Research.
