The EHD3 Knockout A-549 Polyclonal Cells product provides a pooled population of A-549 human lung adenocarcinoma epithelial cells with CRISPR/Cas9-mediated disruption of the EHD3 gene. As a polyclonal knockout model, it comprises a heterogeneous mixture of edited cells, minimizing clonal artifacts and offering a robust loss-of-function system for population-level studies. The use of CRISPR/Cas9 ensures targeted gene disruption, generating a model that retains the native genomic and cellular context of A-549 cells. This product is designed for investigating the functional consequences of EHD3 depletion on endocytic recycling, receptor trafficking, and downstream cellular processes in a well-characterized lung cancer background.
The A-549 cell line was originally established from the lung adenocarcinoma of a 58-year-old Caucasian male and serves as a foundational model for studying alveolar epithelial-derived lung cancer. These cells exhibit an epithelial morphology, express surfactant proteins, and harbor mutations characteristic of lung adenocarcinoma, making them a relevant and widely used host for cancer research. A-549 cells are employed in investigations of oncogenic signaling, drug sensitivity, epithelial-mesenchymal transition, and metastatic behavior. Their utility in knockout models allows researchers to dissect gene function within a clinically pertinent tumor microenvironment-like setting.
EHD3 encodes a dynamin-like ATPase that localizes to endosomes and orchestrates the recycling of internalized receptors to the plasma membrane. It binds phosphatidylinositol-4-phosphate via its EH domain and interacts with Rab11, EHBP1, and F-actin to sort and return cargo such as integrins and VEGFR2. EHD3 cooperates with Arf6 in the endocytic recycling pathway to regulate membrane protrusion formation and cell migration. Upstream, HIF1A transcriptionally activates EHD3 under hypoxia, while growth factors like VEGF and EGF stimulate its activity. Downstream, EHD3-mediated recycling maintains surface VEGFR2 and integrin expression, promoting angiogenic signaling and cell adhesion. Knockout of EHD3 disrupts these processes, reducing receptor availability and impairing migration and angiogenic responses.
In the A-549 lung adenocarcinoma context, loss of EHD3 is anticipated to compromise integrin-dependent adhesion and migration, processes central to tumor invasion and metastatic dissemination. Given the role of VEGFR2 in angiogenic signaling and the responsiveness of A-549 cells to VEGF, this knockout model can be used to dissect how EHD3-mediated receptor recycling contributes to hypoxia-driven angiogenesis and tumor progression. It may also serve as a platform to evaluate the dependency of lung cancer cells on endocytic trafficking for sustained oncogenic signaling and to explore synthetic lethal interactions with existing therapeutics.
Researchers can employ EHD3 Knockout A-549 Polyclonal Cells in a broad range of assays, including western blotting and RT-qPCR to confirm knockout efficiency and assess downstream pathways, immunofluorescence to visualize endosomal morphology, and flow cytometry to quantify surface receptor levels. Functional studies such as Transwell migration, wound-healing, and VEGF stimulation assays are particularly relevant for characterizing the migratory and angiogenic phenotypes. Co-immunoprecipitation can further probe EHD3 interaction networks. This polyclonal knockout product is intended for investigational use in biomedical research. For comprehensive technical details, product specifications, or assistance with experimental design, please contact Ascent Research.
