The GPR75 Knockout T-47D Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt GPR75 expression in the human T-47D breast ductal carcinoma cell line. This heterogeneous pool of edited cells provides a robust loss-of-function model for studying GPR75-dependent signaling and phenotypes without clonal artifacts. The polyclonal format ensures reproducible results across experiments while mitigating selection bias, making it ideal for pooled population analyses.
The parental T-47D cell line, isolated from a pleural effusion of a metastatic mammary adenocarcinoma, is a well-established model for estrogen receptor-positive (ER+) breast cancer. These epithelial cells express estrogen and progesterone receptors, enabling investigations into hormone-responsive pathways and endocrine therapy resistance. Their adherent morphology and stable growth characteristics support a wide array of in vitro functional and biochemical assays.
GPR75 is an orphan G protein-coupled receptor implicated in energy homeostasis and obesity regulation. Upon activation, it couples to G??s protein, stimulating adenylyl cyclase to increase cAMP levels, which activate protein kinase A (PKA) and downstream phosphorylation of the transcription factor CREB. This cascade modulates the expression of genes involved in lipid metabolism. GPR75 also interacts with ??-arrestin and may crosstalk with the MAPK/ERK pathway, though the endogenous ligand remains unknown. Disruption of GPR75 abolishes signaling through the G??s/cAMP/PKA/CREB axis.
In the T-47D background, this knockout model enables dissection of GPR75??s potential roles in cancer cell metabolism and hormone receptor signaling. Given the links between metabolic sensing and tumor progression, the loss-of-function cells can be used to explore whether GPR75 influences proliferation, survival, or metabolic adaptation in ER+ breast cancer. The polyclonal population reflects genetic heterogeneity, enhancing relevance for pharmacological studies and functional genomics.
Key applications include cAMP accumulation assays, PKA activity measurements, CREB phosphorylation analysis by western blot, and RT-qPCR for downstream targets. These cells are also suitable for metabolic flux analysis, proliferation assays, and ligand deorphanization studies in obesity and metabolic syndrome research. Additionally, they can be paired with wild-type controls for genetic rescue or drug response experiments. For technical support or customization inquiries, please contact Ascent Research.
