The EFNA1 Knockout A-549 Polyclonal Cells represent a CRISPR/Cas9-engineered polyclonal cell population derived from the human A-549 lung adenocarcinoma line, with targeted disruption of the EFNA1 gene. This product provides a heterogeneous knockout model that abolishes ephrin-A1 ligand synthesis, enabling rigorous loss-of-function studies in an epithelial cancer background. As polyclonal knockout cells, they offer a pooled genetic perturbation ideal for population-level analyses of ephrin-A1-dependent phenotypes, without the need for monoclonal isolation.
A-549 cells are a classical in vitro model of human alveolar type II epithelium, originally established from a lung adenocarcinoma. These adherent epithelial cells retain key features of the respiratory epithelium and are extensively employed in cancer biology, toxicology, and drug development. Their robust proliferation and well-characterized signaling landscape make them highly suitable hosts for CRISPR-mediated gene editing. The alveolar type II phenotype provides context for investigating tumor cell behaviors that intersect with epithelial differentiation and repair.
EFNA1 encodes ephrin-A1, a glycosylphosphatidylinositol (GPI)-anchored ligand for EphA receptor tyrosine kinases, most notably EPHA2. Ephrin-A1 binding induces receptor forward signaling through FAK (PTK2), ERK2 (MAPK1), and AKT1, while ephrin-A1 reverse signaling via its GPI anchor influences Rho GTPase activity, including RHOA, RAC1, and CDC42, leading to actin cytoskeleton remodeling. Expression of EFNA1 is transcriptionally controlled by HIF1A, EGF, TNF-??, p53, and c-Myc. Additionally, ephrin-A1 interacts with a broad range of EphA receptors (EPHA1-EPHA8, EPHA10) and is susceptible to ectodomain shedding by ADAM10 and ADAM17 metalloproteinases, which modulates signaling output. Disruption of EFNA1 thus simultaneously extinguishes both forward and reverse Eph-ephrin communication, profoundly impacting cellular adhesion, repulsion, and motility pathways.
In A-549 cells, ephrin-A1 is implicated in aggressive tumor behaviors such as migration, invasion, and angiogenesis??hallmarks of lung adenocarcinoma progression. The EFNA1 knockout polyclonal population consequently serves as a powerful tool to dissect the specific contributions of ephrin-A1/EPHA2 signaling to these processes. Researchers can evaluate how loss of ephrin-A1 alters cell-cell contacts, cytoskeletal organization, and downstream signaling cascades, and can assess the model??s utility for studying EphA2-targeted therapeutic strategies in a physiologically relevant lung cancer setting.
This product supports diverse experimental applications, including transwell migration and invasion assays, adhesion studies, and phospho-ERK ELISAs to quantify pathway activity. It is also suited for drug target validation of EPHA2 inhibitors, structure?Cfunction analyses of Eph-ephrin interactions, and whole-genome transcriptional or proteomic profiling. Standard characterization methods such as Western blotting, RT-qPCR, immunofluorescence, and flow cytometry further corroborate knockout efficacy and phenotype. For additional information, please contact Ascent Research.
