The GPR171 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the GPR171 gene has been disrupted. This heterogeneous pool of HAP1 cells provides a loss-of-function model for studying the G protein-coupled receptor GPR171 in biomedical research. The polyclonal format offers a practical and scalable approach for functional genomics studies, enabling rapid screening of receptor-mediated phenotypes without clonal selection.
The parental HAP1 cell line is a near-haploid, adherent cell line derived from the KBM-7 chronic myeloid leukemia cell line of male origin, which harbors the BCR-ABL fusion oncogene. HAP1 cells possess a stable haploid karyotype, making them uniquely suited for CRISPR/Cas9-mediated gene knockout studies, as disruption of a single allele is sufficient to ablate gene function. This genetic simplicity, combined with their robust growth properties and compatibility with a broad range of transfection and assay systems, has established HAP1 as a premier host for targeted genomic engineering and high-throughput functional screening.
GPR171 functions as the cognate receptor for BigLEN, a peptide derived from the proSAAS prohormone. Upon BigLEN binding, GPR171 activates G??i/o proteins, leading to the inhibition of adenylyl cyclase and a subsequent decrease in intracellular cAMP levels. This Gi/o-mediated signaling cascade is central to the regulation of feeding behavior and energy homeostasis. GPR171 also interacts with ??-arrestin, which may mediate receptor desensitization and scaffolding of downstream signaling complexes. The interplay between G??i/o-dependent and ??-arrestin-dependent pathways positions GPR171 as a key sensor in neuropeptide-driven metabolic control.
In the context of HAP1 cells, the knockout of GPR171 provides a clean and defined platform to dissect its signaling mechanisms without interference from other GPCRs that may share downstream effectors. The near-haploid genome ensures that any observed phenotypic changes are directly attributable to the disruption of GPR171, thereby reducing genetic redundancy. This model is particularly valuable for studying Gi/o-coupled receptor pharmacology and for characterizing the functional consequences of ligand?Creceptor interactions in a simplified cellular environment, which can be challenging in diploid or primary cell systems.
Researchers can employ these GPR171 knockout polyclonal cells in a variety of assays to investigate GPCR signaling and metabolic control. cAMP accumulation and calcium flux assays can respectively assess G??i/o-mediated adenylyl cyclase inhibition and G?¦?-mediated signaling. Metabolic profiling, western blotting, RT-qPCR, and immunofluorescence further enable comprehensive phenotypic characterization. These cells are thus invaluable for metabolic disease research, neuropeptide receptor deorphanization, and feeding behavior studies. For additional technical details, please contact Ascent Research.
