The EHD3 Knockout HEK293T Polyclonal Cells offer a versatile CRISPR/Cas9-edited polyclonal knockout cell population for investigating the roles of the endocytic recycling protein EHD3. This product comprises a heterogeneous pool of HEK293T cells with targeted disruption of the EHD3 gene, providing a physiologically relevant loss-of-function model without clonal isolation. The polyclonal format captures diverse genetic lesions, enabling robust population-level analyses of EHD3-dependent processes.
The host cell line, HEK293T, is a transformed human embryonic kidney cell line stably expressing the SV40 large T antigen. Known for its high transfectability and protein expression capacity, HEK293T is extensively used for viral production, signaling studies, and biochemical assays. Its epithelial origin and active membrane trafficking pathways make it an ideal chassis for dissecting endocytic recycling mechanisms.
EHD3 belongs to the EHD ATPase family and functions as a membrane-remodeling protein localized to endocytic recycling compartments. Through its interactions with Arf6, Rab11-FIP2, EHBP1, and Syndapin/PACSIN, EHD3 coordinates the return of internalized receptors, such as EGFR and transferrin receptor, to the plasma membrane. Upstream regulators include Arf6, EGFR signaling, Src kinases, and Rab35, while downstream effects involve Rac1-dependent actin cytoskeleton reorganization. Disruption of EHD3 impairs recycling, leading to altered surface receptor levels and perturbed signaling dynamics.
In the HEK293T context, EHD3 knockout creates a powerful tool to interrogate endocytic trafficking and its crosstalk with signaling networks. The loss of EHD3 is expected to delay EGFR and transferrin receptor recycling, thereby modulating EGFR downstream pathways and actin dynamics. Given HEK293T cells’ well-characterized endosomal system, this model facilitates the study of ciliary protein trafficking and the molecular basis of ciliopathies and cancers, including glioblastoma, where EHD3 dysregulation has been implicated.
Researchers can employ this polyclonal knockout population in a wide range of assays. Western blotting and RT-qPCR confirm EHD3 disruption, while transferrin recycling and EGFR degradation assays quantify functional consequences. Immunofluorescence with endosomal markers reveals trafficking defects, and flow cytometry measures surface receptor levels. Additionally, wound healing assays can assess cell migration influenced by altered actin dynamics. For further information or to acquire this product, please contact Ascent Research.
