The GPR171 Knockout UM-UC-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human UM-UC-3 bladder cancer cell line, with targeted disruption of the GPR171 gene. This polyclonal pool provides a genetically heterogeneous loss-of-function model that avoids clonal artifacts and is well-suited for robust functional analyses in a cancer-relevant context.
The parental UM-UC-3 cell line was originally established from a male patient with transitional cell carcinoma of the bladder and serves as a well-characterized model for invasive urothelial carcinoma. UM-UC-3 cells exhibit high proliferative capacity, mesenchymal-like invasive behavior, and intrinsic chemoresistance, making them particularly useful for investigating metastasis and drug resistance mechanisms in bladder cancer.
GPR171 encodes an orphan receptor activated by the BigLEN peptide, a product of proSAAS processing. Upon ligand binding, GPR171 couples primarily to G??i/o proteins to inhibit adenylyl cyclase and reduce intracellular cAMP, with additional coupling to G??s. The receptor interacts with ??-arrestin1/2 and PDZ domain-containing proteins, scaffolding activation of the MAPK/ERK pathway via MEK-mediated phosphorylation of ERK1/2, and modulating the PI3K/AKT/mTOR axis. Key effectors include CREB and c-Fos, regulated through PKA- and ERK-dependent mechanisms, linking GPR171 to nutrient sensing and cell proliferation.
In the UM-UC-3 bladder cancer background, GPR171 knockout abrogates BigLEN-dependent G protein and ??-arrestin signaling, suppressing cAMP production, attenuating ERK1/2 phosphorylation, and reducing CREB and c-Fos activation. This impairs cell proliferation, migration, and metabolic reprogramming, making the model valuable for dissecting GPCR-driven oncogenic signaling in bladder cancer.
These polyclonal knockout cells are suited for GPCR signaling dissection, GPR171 drug target validation, and metabolic cancer studies. Assays include western blot for phospho-ERK and phospho-CREB, cAMP ELISA, RT-qPCR for c-Fos, MTS proliferation, and Transwell assays. RNA-seq can reveal global transcriptomic changes. Contact Ascent Research for support.
