The GPRC5B Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt GPRC5B expression in HeLa cells. This mixed population model enables loss-of-function studies without monoclonal artifacts, ensuring reliable assessment of GPRC5B-dependent signaling and cellular phenotypes.
The HeLa cell line is derived from a human cervical adenocarcinoma and is immortalized by HPV18, which inactivates the tumor suppressor p53, while telomerase activity sustains its indefinite propagation. As an epithelial model, HeLa cells are instrumental in cancer biology, particularly for dissecting HPV-driven oncogenesis, signal transduction, and therapeutic responses. Their comprehensive genomic annotation and ease of genetic manipulation make them a preferred host for targeted gene knockout studies.
GPRC5B encodes an orphan G protein-coupled receptor that is transcriptionally induced by retinoic acid via RAR/RXR heterodimers. It integrates extracellular cues through G protein-dependent and -independent mechanisms, physically associating with G?? subunits, TAS2R bitter taste receptors, arrestins, and RGS proteins. Downstream, GPRC5B influences key signaling nodes: it modulates the MAPK/ERK cascade through Raf/MEK/ERK and the mTOR pathway via PI3K/AKT/mTORC1, affecting the phosphorylation of ERK1/2, AKT, and S6K. Additionally, it regulates cAMP levels and CREB transcriptional activity, thereby controlling gene expression programs linked to proliferation and differentiation.
In the HPV18-positive HeLa context, ablation of GPRC5B disrupts these intricate signaling networks, offering a powerful model to discern its contributions to cellular growth and morphology. The p53 deficiency of HeLa cells allows researchers to explore GPRC5B function independent of p53-mediated apoptosis, providing insights into its role in cervical adenocarcinoma and potentially in other malignancies such as prostate, lung, and ovarian cancers where its expression has been implicated.
This knockout product supports a wide array of experimental techniques: Western blotting for phospho-ERK1/2, phospho-AKT (Ser473), and phospho-S6 ribosomal protein; RT-qPCR and RNA-seq for transcriptomic changes; immunofluorescence and flow cytometry for protein expression and cell cycle analysis; and functional assays including MTT/CCK8 cell viability, transwell migration and invasion, and cAMP ELISA. Luciferase reporter assays can measure transcriptional responses downstream of retinoic acid or serum stimulation, while co-immunoprecipitation enables mapping of GPRC5B interaction partners. These applications are well-suited for studying GPCR signaling, retinoic acid biology, cancer cell proliferation, and drug sensitivity screening. For additional technical details, please contact Ascent Research.
