The GPR171 Knockout 786-O Polyclonal Cells product provides a heterogeneous pool of CRISPR/Cas9-edited renal carcinoma cells with targeted disruption of the GPR171 gene. This polyclonal knockout population consists of a mixture of independently edited cells, offering a robust loss-of-function model without clonal selection. It enables efficient interrogation of GPR171-dependent phenotypes in a genetically varied context, suitable for applications demanding population-level knockout rather than single-cell-derived homogeneity.
The host 786-O cell line is a well-characterized human clear cell renal cell carcinoma (ccRCC) model derived from a primary renal adenocarcinoma. These epithelial cells harbor a biallelic inactivating mutation in the VHL tumor suppressor gene, leading to constitutive stabilization of hypoxia-inducible factors (HIFs) and mimicking the molecular pathology of most sporadic ccRCC cases. The VHL-deficient background makes 786-O cells a physiologically relevant platform for studying signaling pathways that intersect with renal carcinogenesis and metabolic reprogramming.
GPR171 encodes an orphan G protein-coupled receptor selectively activated by the BigLEN peptide, a neuropeptide processed from the proSAAS precursor by PCSK1. Upon ligand binding, GPR171 couples primarily to G??i/o proteins, inhibiting adenylyl cyclase, reducing intracellular cAMP levels, and attenuating downstream PKA activity and CREB phosphorylation. The receptor also engages ??-arrestin-2 to modulate MAPK/ERK signaling, linking it to ERK1/2 phosphorylation cascades. In T cells, GPR171 expression is induced by TCR/CD3 stimulation and IL-2 signaling, and it negatively regulates immune responses by promoting expression of exhaustion markers PD-1 and TIM-3, interacting with the CD3?? chain. Thus, GPR171 sits at the intersection of neuropeptide-driven GPCR signaling, metabolic regulation, and T cell exhaustion checkpoints.
Within the 786-O ccRCC context, GPR171 knockout disrupts potential autocrine or paracrine BigLEN-mediated signaling that may operate in the tumor microenvironment. This allows dissection of how G??i-coupled cAMP/PKA/CREB and ??-arrestin-dependent ERK pathways influence renal cancer cell proliferation, migration, and survival under VHL-null conditions. Moreover, it facilitates investigation of whether GPR171 contributes to immune evasion by modulating PD-1/TIM-3 expression directly on tumor cells or through cross-talk with infiltrating lymphocytes.
Typical research applications include functional analysis of GPR171 in ccRCC, cAMP and phospho-ERK signaling studies, cell proliferation and migration assays, drug target validation, and tumor microenvironment interaction studies using co-culture systems. The polyclonal pool is compatible with standard readouts such as western blotting, RT-qPCR, cAMP ELISA, co-immunoprecipitation, and RNA-seq. For detailed technical support and lot-specific information, please contact Ascent Research.
