The GPRC5C Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human lung adenocarcinoma cell line. This product provides a heterogeneous pool of cells with targeted disruption of the GPRC5C gene, encoding an orphan G protein-coupled receptor. The polyclonal format preserves population diversity while enabling robust loss-of-function studies without requiring single-cell cloning. Through CRISPR/Cas9-mediated gene disruption, researchers can interrogate GPRC5C function in a biologically relevant epithelial background. This model is ideal for investigating the role of GPRC5C in cancer-relevant signaling networks.
The A-549 host cell line was originally established from the lung adenocarcinoma of a 58-year-old Caucasian male. These alveolar basal epithelial cells serve as a widely used model for non-small cell lung cancer (NSCLC) and are employed extensively in studies of tumor biology, drug metabolism, and chemosensitivity. The A-549 line retains key epithelial characteristics and expresses components of EGFR, retinoic acid, and GPCR signaling pathways, making it an appropriate platform for examining GPRC5C-mediated effects. Its tumorigenic properties in xenograft models further support translational cancer research applications.
GPRC5C functions within a complex signaling landscape, integrating inputs from retinoic acid and EGFR to modulate downstream effectors. Upstream, retinoic acid activates nuclear receptors RAR and RXR, which transcriptionally regulate GPRC5C, while EGFR activation promotes downstream cascades. GPRC5C interacts with ??-arrestins and G proteins, including G??s and G??q, linking to adenylyl cyclase?CcAMP?CPKA pathways and calcium mobilization. Downstream, it influences CREB-mediated transcription and regulates cell cycle proteins p21, p27, and cyclin D1, along with apoptotic regulators from the Bcl-2 family. It also modulates oncogenic nodes NF-??B, STAT3, and YAP/TAZ, impacting proliferation, survival, and migration.
Disruption of GPRC5C in A-549 cells perturbs the confluence of EGFR and retinoic acid signaling. Without GPRC5C, EGF-stimulated activation of the RAS?CRAF?CMEK?CERK cascade and the PI3K?CAKT pathway may be altered, leading to changes in cell proliferation and apoptosis as indicated by downstream targets such as phospho-ERK and cleaved caspase-3. This knockout model is particularly pertinent to lung adenocarcinoma research, where aberrant EGFR and retinoic acid signaling often coexist. By removing a receptor that interfaces with both pathways, scientists can dissect how GPRC5C contributes to tumorigenic processes, including migration and invasiveness, which are central to NSCLC progression.
Typical applications include lung cancer biology, GPCR functional studies, and drug target validation. Researchers employ this model in western blotting, RT-qPCR, MTT, Annexin V, Transwell migration, and immunofluorescence assays to assess expression, viability, apoptosis, invasiveness, and protein localization. Phospho-EGFR/ERK analysis and drug sensitivity profiling enable signal transduction and therapeutic response studies. This polyclonal population supports mechanistic research and screening, providing a versatile tool for cancer signaling. For technical details or to discuss custom applications, please contact Ascent Research.
