The GPR3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HAP1 near-haploid human cell line. This product consists of a heterogeneous pool of cells carrying targeted disruptions of the GPR3 gene, generated by CRISPR/Cas9-mediated gene editing. The polyclonal format provides a population-level loss-of-function model suitable for functional genomics and high-throughput phenotypic screening without the need for single-cell cloning.
HAP1 cells originate from the KBM-7 chronic myeloid leukemia line and maintain a near-haploid karyotype, making them an established platform for genetic screens and reverse genetics. Their haploid nature facilitates unambiguous knockout phenotypes because a single gene disruption generally abrogates function, and the fibroblast-like morphology supports robust culturing and assay development. HAP1 cells are widely employed in haploid genetic screens and functional genomics due to their ease of manipulation and predictable genetics.
GPR3 encodes a constitutively active Gs-coupled orphan GPCR that drives cAMP production through adenylyl cyclase activation. Downstream, cAMP activates protein kinase A (PKA), which phosphorylates targets including CREB1 to modulate transcription. GPR3 also enhances gamma-secretase-mediated processing of APP, increasing amyloid-beta production, and interacts with GNAS, beta-arrestin-2, and caveolin-1. Cholesterol is a proposed modulator of receptor activity, linking this pathway to broader GPCR signaling networks. This constitutive activity regulates key processes including oocyte meiotic arrest and neuronal differentiation.
In the HAP1 near-haploid context, GPR3 disruption eliminates receptor function in the majority of cells, creating a clean background for studying GPR3-dependent cAMP-PKA-CREB and gamma-secretase pathways. This model is particularly advantageous for investigating the receptor’s roles in oocyte meiotic arrest, neuronal differentiation, and Alzheimer’s-related amyloid pathology, and it simplifies complementation rescue experiments with GPR3 expression vectors. The haploid setting also minimizes interference from functional redundancy, enabling clearer interpretation of signaling phenotypes.
This polyclonal knockout is ideal for cAMP accumulation assays, phospho-CREB Western blotting, amyloid-beta ELISA, and gamma-secretase activity measurements. Representative assays also include RT-qPCR and complementation with wild-type GPR3 to confirm on-target effects. These cells support functional genomics screens, GPCR drug target validation, and mechanistic studies on constitutive receptor signaling in Alzheimer’s disease, reproductive disorders, and cancer. For additional technical specifications or support, please contact Ascent Research.
