The GPRC5C Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population of the human HAP1 cell line, designed for targeted disruption of the GPRC5C gene. Supplied as a polyclonal pool, this heterogeneous loss-of-function model enables functional genomics studies without clonal expansion artifacts. CRISPR-mediated gene disruption ablates GPRC5C expression, allowing investigation of its signaling roles in a near-haploid background.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myelogenous leukemia line, retaining a haploid chromosome complement except for a small disomic region. Its haploid genome simplifies genotype-phenotype correlation in CRISPR knockout screens, as single-copy gene disruption results in complete loss of function. The line exhibits stable growth and preserved signaling competence, serving as a robust platform for cancer-related pathway analysis.
GPRC5C is an orphan class C GPCR transcriptionally regulated by all-trans retinoic acid via RAR/RXR and SP1. It is predicted to interact with heterotrimeric G proteins and ??-arrestins. Functional evidence suggests it acts as a tumor suppressor by inhibiting NF-??B (p65/p50) transcriptional activity, attenuating STAT3 phosphorylation, and suppressing ??-catenin-mediated Wnt signaling. Downstream, it modulates calcium/calmodulin-dependent kinases, linking retinoic acid signals to control of proliferation, apoptosis, and migration.
In the HAP1 polyclonal knockout population, GPRC5C disruption permits direct measurement of changes in NF-??B, STAT3, and ??-catenin pathway activity without allele redundancy. The haploid background ensures functional knockout in most cells, enhancing signal-to-noise in biochemical and phenotypic assays. This model is particularly suited for defining GPRC5C??s role in negative regulation of oncogenic signaling and for exploring its relevance to non-small cell lung cancer, colorectal cancer, gastric cancer, and metabolic disorders linked to retinoic acid sensing.
Applications include functional genomics screens, cancer signaling studies, drug target validation, and GPCR biology. Researchers can perform Western blotting, RT-qPCR, NF-??B reporter assays, STAT3 phospho-analysis, cell proliferation, migration, and invasion assays, and RNA-seq to map GPRC5C-dependent networks. Combinatorial treatments with retinoic acid or pathway inhibitors may further model therapeutic interventions. For further technical information or custom projects, please contact Ascent Research.
