The EHD4 Knockout A-549 Polyclonal Cells product provides a heterogeneous population of A-549 cells with targeted disruption of the EHD4 gene via CRISPR/Cas9-mediated gene editing. This polyclonal knockout pool serves as a robust loss-of-function model to study endocytic trafficking in a human lung adenocarcinoma background. Supplied as a cryopreserved vial, the population avoids clonal selection biases, enabling researchers to investigate collective consequences of EHD4 ablation with representation of diverse genetic outcomes.
The parental A-549 cell line, derived from a 58-year-old Caucasian male lung adenocarcinoma, is a widely used model in cancer and respiratory research. These alveolar type II epithelial-like cells retain key oncogenic signaling networks, including EGFR pathways, making them highly relevant for dissecting receptor trafficking dynamics in a cancer context. Their established experimental workflows facilitate detailed investigations of tumorigenesis, metastasis, and drug response.
EHD4 encodes a membrane tubulating protein that functions in endosomal recycling, orchestrating the return of internalized receptors and adhesion molecules to the plasma membrane. It operates downstream of Arf6 and PIP2 and in concert with Rab11, Rab5, and effectors such as Rab11-FIP2 and syndapin. EHD4 directly recycles cargoes including transferrin receptor (TfR) and EGFR, and its activity is modulated by EGF/EGFR signaling. Interactions with amphiphysin, actin, and cytoskeletal regulators further link endocytic traffic to cell migration and cytokinesis.
In the A-549 adenocarcinoma background, EHD4 disruption impairs endosomal recycling, altering surface expression and trafficking of receptors like EGFR and TfR. This perturbation enables dissection of how recycling defects influence oncogenic signaling, adhesion, and migration. Given the role of dysregulated receptor trafficking in lung adenocarcinoma progression, this polyclonal knockout model reflects tumor heterogeneity and provides a powerful platform to examine EHD4’s contribution to malignant phenotypes.
This model supports diverse assays including transferrin uptake and recycling kinetics, EGFR degradation analysis, Transwell migration/invasion assays, and immunofluorescence-based receptor localization studies. Additional applications encompass phospho-EGFR signaling analysis, flow cytometry for surface receptor quantification, co-immunoprecipitation of protein interactions, and RNA-seq transcriptomic profiling. These approaches facilitate investigation of compensatory pathways, drug response profiling, and the role of EHD4 in metastasis. For further technical inquiries, please contact Ascent Research.
