EHD4 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the EHD4 gene in a HeLa cell background. This polyclonal pool provides a heterogeneous loss-of-function model, enabling robust investigation of EHD4-dependent cellular processes without the selection of a single clonal isolate. The use of CRISPR/Cas9-mediated gene disruption ensures efficient targeting of the EHD4 locus, resulting in a mixed population that retains the diverse genetic background of the host cells while eliminating functional EHD4 expression across the population.
HeLa cells are an immortalized human cervical adenocarcinoma epithelial cell line originally derived from Henrietta Lacks. They are widely utilized in biomedical research due to their robust proliferative capacity, extensive characterization, and relevance as a model for human cell biology and cancer. Their epithelial origin and transformed phenotype make them particularly suitable for studying processes such as cell adhesion, migration, and receptor trafficking, all of which are intimately linked to EHD4 function.
EHD4 encodes an ATPase that is a critical regulator of endosomal trafficking, specifically facilitating receptor recycling to the plasma membrane. It functions in the endocytic recycling pathway by promoting membrane curvature and scission on Rab11-positive recycling endosomes. EHD4 activity is activated downstream of EGF receptor signaling and Arf6 GTPase, and it works in concert with Rab11, EHD1, and the scaffolding proteins MICAL-L1 and Rab11-FIP2. Its primary downstream effects include the recycling of ??1 integrin and transferrin receptor, which in turn modulates FAK signaling and actin cytoskeleton remodeling, thereby influencing cell adhesion, migration, and signaling cascades.
In the HeLa cell context, disruption of EHD4 offers a powerful model to dissect its role in cancer-associated phenotypes. Given the well-documented connection between endosomal recycling and cancer metastasis, these polyclonal knockout cells are particularly valuable for studying how EHD4-dependent trafficking of integrins and other receptors contributes to tumor cell migration and invasion. The model enables functional interrogation of EHD4 in an epithelial cancer background, providing insights that may be relevant to cancers such as breast cancer and glioblastoma.
Typical research applications include the investigation of endocytic recycling mechanisms, analysis of receptor trafficking dynamics during cell migration, and functional studies of EHD4 in tumor progression. Representative assays that can be performed with these cells encompass transferrin uptake/recycling assays, integrin recycling by flow cytometry, wound healing migration assays, Transwell invasion assays, and immunofluorescence microscopy for Rab11-positive recycling endosomes. Gene disruption can be validated by Western blotting for EHD4 protein and RT-qPCR for EHD4 mRNA. This product is ideal for researchers aiming to elucidate the molecular underpinnings of receptor recycling and its impact on cancer cell behavior. For further details or technical inquiries, please contact Ascent Research.
