The EHD2 Knockout HeLa Polyclonal Cells product provides a heterogeneous population of HeLa cells that have undergone CRISPR/Cas9-mediated gene disruption targeting the EHD2 locus, generating a polyclonal knockout model. This population retains the inherent genetic variability arising from polyclonal editing and serves as a robust loss-of-function system for probing EHD2-dependent biological processes without the selection of single-cell clones. The knockout format ensures a representative background for studies requiring physiological complexity, making it suitable for a wide range of functional assays in cell biology and cancer research.
HeLa cells, the host line for this model, are derived from a human cervical adenocarcinoma and are positive for human papillomavirus type 18 (HPV-18). As an immortalized epithelial cell line, HeLa exhibits rapid proliferation, transformed morphology, and well-characterized signaling networks, including integrin-mediated adhesion and caveolae-dependent endocytosis. These characteristics render HeLa cells a widely employed platform for investigating cancer cell invasion, membrane trafficking, and cytoskeletal dynamics, with extensive literature supporting their utility in mechanistic and drug discovery studies.
EHD2 encodes an ATP-dependent membrane-remodeling ATPase that stabilizes caveolae and regulates the endocytic recycling of integrins, particularly integrin beta1. Activation by upstream cues such as integrin ligation, caveolin-1, and mechanical stress promotes EHD2 interaction with caveolin-1 and PACSIN2, orchestrating caveolar coat assembly. EHD2 functions downstream of integrin engagement and upstream of Rac1-mediated actin polymerization, modulating focal adhesion turnover and cell migration. Key pathway components influenced by EHD2 include caveolin-1, PTRF/Cavin1, integrin beta1, focal adhesion kinase (FAK), and Src kinase, highlighting its central role in mechanotransduction and adhesion signaling.
In HeLa cells, which maintain functional caveolar structures and integrin trafficking machinery, disruption of EHD2 is expected to impair caveolar stability and integrin beta1 recycling, leading to altered cell adhesion, defective mechanosensing, and reduced migratory capacity. This knockout model thus enables precise dissection of EHD2??s contribution to cancer cell invasion and metastasis, and provides a relevant context for studying cardiovascular disorders where caveolae function is essential. The loss of EHD2 may also compromise endocytic recycling pathways critical for membrane homeostasis and signal transduction.
Researchers can employ this model in diverse experimental settings: Western blotting to confirm EHD2 depletion and assess caveolin-1 levels; immunofluorescence to visualize caveolar integrity and focal adhesion dynamics; and Boyden chamber assays to quantify changes in migration and invasion. Integrin recycling assays using antibody-uptake protocols, co-immunoprecipitation studies with EHD2-caveolin-1 complexes, and GTPase activation assays for Rac1 facilitate in-depth mechanistic analyses linking EHD2 to actin cytoskeleton remodeling. This knockout product is ideal for caveolae biology, cancer invasion research, membrane trafficking investigations, and drug delivery studies. For further details or custom requests, please contact Ascent Research.
