GPRC5A Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the A-549 human lung adenocarcinoma cell line. These cells carry a targeted disruption of the GPRC5A gene, an orphan G protein-coupled receptor with tumor-suppressive functions. The polyclonal knockout format reflects a heterogeneous pool of edited cells, enabling population-level loss-of-function studies without clonal selection artifacts. This product is suitable for examining GPRC5A-dependent signaling in a disease-relevant epithelial background.
The parental A-549 cell line originates from the lung adenocarcinoma tissue of a 58-year-old Caucasian male. These adherent epithelial cells are widely used as a model for alveolar type II epithelium and lung adenocarcinoma biology. A-549 cells retain key features of transformed lung epithelium, including aberrant proliferative signaling and migratory capacity, making them a standard host for oncogene and tumor suppressor gene studies. The knockout of GPRC5A in this context allows investigation of receptor-mediated regulatory networks within a well-characterized respiratory disease model.
GPRC5A functions as an orphan GPCR tumor suppressor, negatively regulating cell proliferation, differentiation, and apoptosis. Mechanistically, GPRC5A is activated by all-trans retinoic acid (ATRA) and synthetic retinoids such as tazarotene through retinoic acid receptors (RARs). It interacts with components of the Wnt/??-catenin pathway, including Frizzled receptors and Dishevelled, and associates with the ??-catenin destruction complex. Downstream, GPRC5A suppresses ??-catenin/TCF/LEF transcriptional activity, reducing expression of pro-proliferative targets like Cyclin D1. Additionally, GPRC5A inhibits NF-??B signaling by limiting p65 nuclear translocation and transcriptional activity, thereby repressing MMP-9 and Survivin. Retinoic acid-mediated activation of GPRC5A also attenuates STAT3 and EGFR signaling cascades, underscoring its integrative role in constraining oncogenic networks.
In A-549 cells, GPRC5A is frequently silenced by promoter methylation in lung adenocarcinoma, and genetic knockout replicates this epigenetic loss-of-function state. Disruption of GPRC5A unleashes Wnt/??-catenin and NF-??B pathways, driving increased proliferation, enhanced migration, and reduced epithelial differentiation??hallmarks of aggressive tumor behavior. The polyclonal GPRC5A knockout A-549 model thus recapitulates critical aspects of tumor progression observed in non-small cell lung cancer and serves as a valuable tool for dissecting GPRC5A-dependent suppression of these signaling axes in a relevant cellular milieu.
Researchers can employ this knockout model for diverse applications, including cancer biology, tumor suppressor gene studies, and lung adenocarcinoma research. Experimentally, confirmation of GPRC5A disruption via Western blotting and RT-qPCR is recommended, followed by functional assays such as ??-catenin/TCF luciferase reporter assays, MTT proliferation assays, Transwell migration/invasion assays, and Annexin V apoptosis analyses. Immunofluorescence for ??-catenin localization further clarifies pathway activation status. These approaches enable detailed investigation of retinoid-based therapies, epithelial differentiation mechanisms, and metastasis. For additional product details or technical support, contact Ascent Research.
