The ADGRA3 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population from the A-549 human lung adenocarcinoma cell line, with targeted disruption of ADGRA3. This loss-of-function model avoids clonal selection artifacts, preserving heterogeneous editing events that better reflect tumor cell diversity.
The parental A-549 cell line originates from human lung adenocarcinoma epithelium and is a standard in vitro model for non-small cell lung cancer. These adherent cells are wild-type for p53 and carry a KRAS G12S mutation, key oncogenic features. This genetic background is ideal for studying adhesion-mediated migration and invasion in a lung adenocarcinoma context.
ADGRA3 is an adhesion GPCR that couples cell adhesion to Wnt/planar cell polarity (PCP) signaling. Extracellular matrix components and mechanical stimuli activate ADGRA3, leading to recruitment of DVL and ??-arrestin, and subsequent G protein-dependent and independent signaling. ADGRA3 promotes RhoA activation, which stimulates ROCK-mediated actin cytoskeletal reorganization and JNK phosphorylation; it also stabilizes ??-catenin, linking physical cues to transcriptional programs that govern directed cell migration and invasive behavior.
In A-549 cells, ADGRA3 knockout disrupts adhesion-dependent Wnt/PCP signaling and impairs RhoA-ROCK-driven cytoskeletal remodeling, resulting in diminished migration and invasion. The mechanistic model posits that loss of ADGRA3 decouples extracellular adhesive inputs from the DVL?CRhoA?CROCK?CJNK signaling cascade and ??-catenin dynamics. Given the prevalence of KRAS mutations in lung adenocarcinoma, this knockout model is particularly useful for dissecting how adhesion receptor signaling modulates KRAS-dependent phenotypes and for evaluating sensitivity to pathway inhibitors.
Typical experimental applications include wound healing assays to assess collective migration, transwell migration and invasion assays to quantify directional motility and matrix degradation, and adhesion assays on fibronectin or collagen matrices. Molecular readouts such as western blotting for phospho-JNK and ??-catenin, immunofluorescence staining of actin filaments, and RNA-seq transcriptomic profiling provide mechanistic validation. These cells are a powerful tool for cancer metastasis research, GPCR-targeted therapy investigations, and functional genomics studies of adhesion receptors. For further information, please contact Ascent Research.
