The GPR75 Knockout MCF-7 Polyclonal Cells are a polyclonal population generated by CRISPR/Cas9-mediated gene disruption of GPR75 in the MCF-7 human breast adenocarcinoma cell line. This pooled knockout model enables the study of GPR75 function without the constraints of a single clonal derivative, offering a broad representation of editing outcomes while maintaining a consistent loss-of-function phenotype suitable for robust functional analyses.
The parental MCF-7 cell line was originally isolated from the pleural effusion of a 69-year-old Caucasian female with metastatic breast cancer. These cells are estrogen receptor-positive (ER+), progesterone receptor-positive (PR+), and responsive to hormonal stimulation, making them a well-established in vitro model for hormone receptor-positive breast cancer. MCF-7 cells retain key features of luminal A breast cancer and are widely employed in studies of hormone signaling, therapeutic resistance, and cancer cell biology.
GPR75 encodes an orphan G protein-coupled receptor that couples to G??i and G??q heterotrimeric G proteins, leading to modulation of adenylate cyclase and phospholipase C. This receptor activity alters intracellular levels of cAMP and inositol trisphosphate (IP3), thereby influencing protein kinase A (PKA) and protein kinase C (PKC) signaling. Downstream, GPR75 activates the PI3K/Akt and MAPK/ERK cascades, with key molecular targets including Akt, ERK, and the transcription factor CREB. ??-arrestin recruitment further modulates GPR75 signaling and receptor trafficking. Through these pathways, GPR75 has been implicated in neuroprotection, energy metabolism, and potentially in the survival and proliferation of cancer cells.
In the MCF-7 breast cancer context, disruption of GPR75 provides a valuable tool to dissect the receptor??s contribution to hormone receptor-positive tumor biology. Activation of the PI3K/Akt and MAPK/ERK pathways is frequently dysregulated in breast cancer and drives cell growth, migration, and metabolic reprogramming. By abrogating GPR75-mediated signaling, researchers can investigate its role in estrogen-dependent and estrogen-independent proliferation, assess its impact on downstream effectors such as phosphorylated Akt and ERK, and explore crosstalk between orphan GPCRs and steroid hormone signaling. This model also facilitates the study of GPR75 in metabolic syndrome and obesity-related cancer risk, given the receptor??s dual roles.
Typical applications of these GPR75 knockout polyclonal MCF-7 cells include detailed mechanistic studies using Western blotting, RT-qPCR, and transcriptomic profiling (RNA-seq) to validate loss of GPR75 and monitor downstream pathway alterations. Functional assays such as MTT proliferation, transwell migration, and Annexin V apoptosis assays enable quantitative assessment of cellular phenotypes. Measurement of intracellular cAMP levels, phospho-Akt ELISA, and calcium flux assays further characterize signaling perturbations. These cells are also suitable for ligand screening and therapeutic target validation in cancer and metabolic disease. For additional product details or technical support, please contact Ascent Research.
