EHD4 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human EHD4 gene in the NCI-H1975 epithelial cell line. This product provides a heterogeneous pool of loss-of-function cells generated through targeted gene disruption, enabling robust functional studies of EHD4-dependent endocytic trafficking without clonal selection. The polyclonal format reflects the natural diversity of genomic editing outcomes across the cell population, making it suitable for experiments where pooled knockout effects average out clonal variability.
The host cell line NCI-H1975 is a widely utilized human non-small cell lung cancer (NSCLC) model, originally derived from a female patient with lung adenocarcinoma. As an epithelial cell line, NCI-H1975 retains key characteristics of adenocarcinoma, including dependency on oncogenic signaling networks and sensitivity to targeted therapies. It is extensively employed in cancer biology research to dissect mechanisms of proliferation, survival, migration, and drug resistance, particularly in the context of EGFR-driven malignancies.
EHD4 encodes a dynamin-related ATPase that functions at the nexus of clathrin-mediated endocytosis and actin remodeling. EHD4 is activated downstream of EGF and EGFR and operates in concert with RAB GTPases such as Rab5 and Rab11. It interacts with Syndapin-1, Amphiphysin-1, Clathrin, AP-2, and Rab11-FIP2 to facilitate scission of clathrin-coated vesicles and promote recycling of internalized cargo, including EGFR and integrins, back to the plasma membrane. By coupling receptor recycling to F-actin cytoskeletal reorganization via ARF6 and PIP2, EHD4 modulates cell adhesion and migration dynamics.
In the NCI-H1975 background, disruption of EHD4 is expected to impair clathrin-mediated internalization and intracellular sorting of EGFR, thereby attenuating downstream signaling cascades that drive tumor cell proliferation, survival, and motility. This loss-of-function model enables researchers to interrogate how endocytic trafficking influences oncogenic signaling and therapeutic sensitivity. The polyclonal knockout population provides a physiologically relevant reduction in EHD4 activity, avoiding potential clonal artifacts while maintaining the cellular heterogeneity characteristic of tumor biology.
This product is suited for a range of targeted investigations, including the study of EGFR trafficking kinetics in NSCLC, the contribution of endocytic pathways to acquired drug resistance, and functional dissection of EHD4-dependent cell migration. Representative assays include Western blotting to monitor EGFR and downstream signaling effectors, immunofluorescence to visualize receptor internalization and recycling defects, flow cytometry for quantification of surface receptor levels, and co-immunoprecipitation to assess protein-protein interactions within the endocytic machinery. Migration and apoptosis assays under drug treatment further enable evaluation of EHD4??s role in therapeutic response. For further details or to place an order, please contact Ascent Research.
