The EHD2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T host line, designed to disrupt the EHD2 gene. This gene-edited product provides a loss-of-function model for investigating the roles of EHD2 in endocytic recycling and intracellular membrane trafficking without specifying the editing pattern or clonal origin. The polyclonal knockout pool enables robust assessment of EHD2-dependent processes in a human embryonic kidney epithelial background for mechanistic and phenotypic studies.
The host cell line, HEK293T, is a human embryonic kidney epithelial cell derivative that stably expresses the SV40 large T antigen, enabling high-titer viral production and efficient transient protein expression. Widely employed for gene expression, signal transduction analysis, and viral packaging, HEK293T cells offer a genetically tractable and well-characterized platform for interrogating gene function. Their epithelial origin provides a relevant cellular context for studying membrane trafficking pathways such as caveolae-mediated endocytosis.
EHD2 functions as a key regulator of caveolae-dependent endocytosis and endocytic recycling, linking membrane receptor trafficking to cytoskeletal dynamics. It is activated downstream of integrin signaling and Src kinase, with inputs from EGFR and PtdIns(4,5)P2. EHD2 interacts with caveolin-1 and cortactin and forms complexes with F-actin to stabilize caveolae and direct actin remodeling. Downstream effects modulate trafficking of integrins (including ??1) and caveolin-1, influencing pathways involving talin, dynamin, and cavin-1.
In HEK293T cells, disruption of EHD2 is expected to perturb caveolae stability and alter the endocytic routing of integrins and other cargo, leading to defined defects in cell adhesion, spreading, and migration. The loss of EHD2-mediated regulation may also impact EGFR recycling and signaling output, providing a model to dissect how endocytic trafficking influences downstream kinase cascades. Because HEK293T cells are permissive for viral transduction and transient overexpression, this polyclonal knockout pool can be readily complemented with mutant or wild-type EHD2 constructs to validate phenotype?Cgenotype relationships in a human epithelial setting.
This EHD2 knockout model supports applications in caveolae biology, endocytic trafficking, cancer metastasis research, and drug screening for endocytosis modulators. Key assays include Western blotting, RT-qPCR, flow cytometry, immunofluorescence, co-immunoprecipitation, migration and invasion assays, and endocytosis assays. For further technical specifications and ordering information, please contact Ascent Research.
