GPR171 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered for loss-of-function studies of the GPR171 gene. Generated via CRISPR/Cas9-mediated gene disruption in HeLa cells, this polyclonal pool provides a versatile model for investigating GPR171-dependent signaling without the need for clonal isolation. The heterogeneous knockout population enables robust functional assays while reducing clone-specific variability.
The HeLa cell line, an HPV18-positive human cervical adenocarcinoma, is a well-established model in cancer research and signal transduction. Its well-characterized signaling pathways and transfection efficiency make it ideal for studying GPCR function. HeLa cells express many components of the G protein-coupled receptor signaling machinery, offering a physiological platform to examine GPR171-mediated neuropeptide responses relevant to feeding and energy regulation.
GPR171 is a Gi/o-coupled receptor activated by the Big LEN neuropeptide, derived from the ProSAAS precursor. Ligand binding triggers G??i/o-mediated inhibition of adenylyl cyclase, lowering cAMP levels and subsequently reducing PKA activity and modulating ERK1/2 phosphorylation. The receptor also interacts with beta-arrestin and GRKs to regulate desensitization and trafficking. This signaling cascade connects extracellular cues to intracellular pathways controlling energy homeostasis and feeding behavior.
In the HeLa background, disruption of GPR171 allows researchers to interrogate the receptor??s role in cAMP and ERK signaling. The loss of Big LEN-induced adenylyl cyclase inhibition provides a clear readout for functional assays. This knockout model is particularly suited for metabolic and obesity research, as it enables the study of neuropeptide receptor signaling in a tractable cell line with relevance to energy balance pathways.
These polyclonal knockout cells are applicable in cAMP accumulation assays, phospho-ERK Western blotting, and RT-qPCR for gene expression profiling. Beta-arrestin recruitment and co-immunoprecipitation studies can dissect receptor coupling and trafficking. Immunofluorescence and flow cytometry enable localization and expression analysis. The cells are valuable for GPCR drug target validation and neuropeptide signaling research. For technical inquiries, please contact Ascent Research.
