The EHD4 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited human cell population carrying targeted disruption of the EHD4 gene. This polyclonal knockout model provides a biologically relevant loss-of-function system for investigating EHD4-dependent processes without clonal selection, preserving the heterogeneity of the edited pool.
The host cell line, Huh-7, is a well-characterized epithelial cell line derived from a human hepatocellular carcinoma. Huh-7 cells retain many hepatocyte features and are extensively used in liver cancer research, hepatic biology studies, and the analysis of endocytic trafficking and signal transduction. Their robust growth and well-defined molecular landscape make them an ideal background for genetic perturbation studies in hepatocellular carcinoma.
EHD4 is a member of the C-terminal EH domain-containing protein family and functions as a key regulator of endocytic recycling and receptor internalization. It dynamically interacts with actin filaments and associates with membrane compartments, including early and recycling endosomes. EHD4 directly binds to and cooperates with dynamin, Rab GTPases, and other EHD family proteins to orchestrate membrane remodeling. Signaling inputs from the epidermal growth factor receptor (EGFR) activate EHD4-dependent pathways, leading to the modulation of downstream effectors such as MAPK and Akt, as well as the trafficking of integrins. Through these interactions, EHD4 couples actin cytoskeleton dynamics to receptor sorting, thereby controlling the spatial and temporal output of cell surface receptor-mediated signals.
In the hepatocellular carcinoma setting, EHD4-mediated endocytic trafficking contributes to the regulation of oncogenic signaling pathways and cellular behaviors including proliferation, migration, and adhesion. Loss of EHD4 in Huh-7 cells can perturb EGFR recycling and downstream MAPK/Akt activity, potentially altering integrin-dependent adhesion and providing a model to dissect how endosomal sorting influences cancer cell phenotype. This knockout model enables researchers to specifically interrogate the role of EHD4 in liver cancer biology and to explore its contribution to pathological endocytic dysregulation.
The EHD4 Knockout Huh-7 Polyclonal Cells are a versatile tool for a range of advanced research applications. They are suitable for receptor internalization and recycling assays using fluorescent ligands or antibody-feeding protocols, quantitative analysis of signaling kinetics via Western blotting and flow cytometry, and immunofluorescence co-localization studies of endosomal markers and actin. This model also supports transcriptomic analysis by RT-qPCR to assess gene expression changes downstream of EHD4 disruption. By combining biochemical and cell-based readouts, researchers can delineate the molecular mechanisms coupling EHD4 to membrane trafficking and signal transduction in hepatocellular carcinoma. For technical inquiries and ordering information, please contact Ascent Research.
