EHD2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the EHD2 gene is disrupted, providing a loss-of-function model for investigating endocytic recycling and caveolar biology. This polyclonal population encompasses a spectrum of genetic alterations, enabling robust functional studies without clonal selection. The knockout impairs EHD2-mediated membrane tubulation and receptor trafficking, offering researchers a versatile tool to dissect EHD2-dependent processes in a human epithelial context.
The host HT29 cell line, derived from a 44-year-old female with colorectal adenocarcinoma, displays epithelial morphology and forms polarized monolayers, serving as a well-established model of intestinal epithelial biology and colorectal cancer. These cells are widely employed to study endocytosis, cell polarity, and tumor migration, and their characterized signaling landscape provides an ideal background for examining how EHD2 disruption affects adhesion and trafficking pathways.
EHD2 is an ATPase that binds lipid membranes to facilitate tubulation, functioning downstream of transcriptional regulators SRF and Myocardin and under the control of Caveolin-1 and integrin signaling. It interacts with Caveolin-1, EHD1, EHD3, EHD4, Actin, and Intersectin-2 to coordinate focal adhesion turnover and the recycling of receptors such as the Transferrin receptor. Within the caveolar network, EHD2 operates alongside Dynamin-2, Rab5, and Rab11, and its activity modulates Integrin beta1, FAK, and Src signaling, linking endocytic trafficking to cell adhesion and motility.
In HT29 cells, loss of EHD2 is predicted to impair receptor recycling and focal adhesion dynamics, leading to altered migration and invasion??processes central to colorectal cancer progression and metastasis. This knockout model enables detailed dissection of EHD2’s contribution to caveolar organization and integrin-mediated adhesion, revealing how disrupted recycling impacts downstream signaling through FAK and Src and influences cell spreading and polarity in a tumor-relevant setting.
Typical applications include Western blotting and immunofluorescence to verify EHD2 ablation and assess Caveolin-1 and focal adhesion markers, Transferrin uptake and recycling assays to measure endocytic trafficking efficiency, and migration and invasion assays in modified Boyden chambers to evaluate metastatic potential. Co-immunoprecipitation studies with Caveolin-1 probe disrupted interactions, while RT-qPCR quantifies integrin and EHD2 transcript levels and cell spreading assays on fibronectin reveal adhesion defects. For further information, please contact Ascent Research.
