The GPR171 Knockout KYSE-30 Polyclonal Cells provide a CRISPR/Cas9-mediated gene-disrupted population targeting the GPR171 locus in the KYSE-30 human esophageal squamous cell carcinoma line. This polyclonal knockout pool enables loss-of-function studies of the G protein-coupled receptor 171 (GPR171) within a heterogeneous cellular context, avoiding clonal artifacts and facilitating robust assessment of gene function across mixed genetic backgrounds. The cells are designed for researchers investigating GPCR signaling, metabolic regulation, and oncogenic processes.
KYSE-30 is an established epithelial tumor cell line derived from a well-differentiated invasive esophageal squamous cell carcinoma of a 64-year-old male patient. The line harbors KRAS amplification and expresses epidermal growth factor receptor (EGFR), reflecting key molecular features of aggressive esophageal carcinoma. Its well-differentiated phenotype and growth characteristics make it a relevant model for studying tumor cell invasion, proliferation, and signal transduction in esophageal cancer research.
GPR171 functions as a Gi/o-coupled receptor activated by the BigLEN peptide, a product of ProSAAS processing by prohormone convertases PCSK1 and PCSK2. Upon ligand binding, GPR171 engages G??i/o proteins to inhibit adenylyl cyclase, leading to decreased intracellular cAMP levels and attenuated protein kinase A (PKA) activity. Concurrently, GPR171 signaling stimulates the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway, with ??-arrestin1/2 and GPCR kinases (GRKs) serving as interacting regulators. In this signaling network, BigLEN acts upstream of GPR171, while adenylyl cyclase, cAMP, PKA, and ERK1/2 function as key downstream effectors.
In the KYSE-30 background, disruption of GPR171 eliminates the receptor-mediated inhibition of adenylyl cyclase, thereby elevating basal cAMP levels and altering PKA and ERK1/2 signal transduction. Given the invasive nature of the parental line and its KRAS/EGFR status, GPR171 knockout may perturb tumor cell proliferation, migration, and metabolic adaptation, providing a unique system to dissect the contribution of nutrient- and energy-sensing GPCR pathways to esophageal cancer progression. The polyclonal format preserves the cellular heterogeneity inherent in tumor biology, offering a more physiologically relevant model than clonal isolates.
Researchers can employ this knockout model in a variety of functional assays, including cAMP accumulation measurements to assess adenylyl cyclase regulation, western blot analysis of ERK1/2 phosphorylation, MTT and colony formation assays for proliferation, and Transwell migration/invasion assays. Additional applications encompass flow cytometry-based apoptosis studies and qPCR profiling of downstream transcriptional changes. This product is suitable for drug target validation, exploration of metabolic regulation in cancer, and dissection of GPCR-mediated signaling in tumorigenesis. For further information, please contact Ascent Research.
