The EHD2 Knockout Huh-7 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the EHD2 gene in the human Huh-7 hepatocellular carcinoma cell line. This polyclonal pool contains a heterogeneous cell population carrying gene disruptions introduced by CRISPR/Cas9, creating a loss-of-function model that avoids clonal selection artifacts. The product is designed for immediate use in functional studies requiring EHD2 deficiency.
Huh-7 is a well-differentiated human hepatocellular carcinoma cell line established from a 57-year-old Japanese male, characterized by mutant p53. It is widely used to study liver function, hepatitis C virus (HCV) infection, and hepatocellular carcinoma biology. This adherent epithelial line retains hepatocyte features and is permissive to HCV entry and replication, making it a standard platform for antiviral and hepatocarcinogenesis research.
The EHD2 gene encodes an ATPase that stabilizes caveolae at the plasma membrane through interactions with CAV1, PACSIN2, and F-actin. It is regulated by upstream signals such as CAV1, TGF-??1, Notch1, integrin adhesion, and hypoxia. Downstream, EHD2 promotes RhoA activation, Rac1 inhibition, and FAK/Src signaling, controlling actin stress fiber formation and focal adhesion dynamics. Knockout disrupts caveolae, leading to dysregulated endocytic recycling and altered cell migration.
In Huh-7 cells, EHD2 knockout allows dissection of caveolae-dependent processes critical for hepatocellular carcinoma metastasis, insulin receptor trafficking, and HCV entry. Loss of EHD2 impairs promigratory signaling and actin reorganization, providing a tool to study how caveolae disassembly influences tumor cell invasion. Additionally, it enables investigation of metabolic dysregulation and viral host membrane interactions in a liver cancer context.
Researchers can utilize these cells for caveolae-mediated endocytosis studies via transferrin uptake assays and electron microscopy. Metastasis research is supported by wound healing and Boyden chamber assays, with phospho-signaling analysis of FAK/Akt. Insulin signaling and HCV entry mechanisms can be assessed using immunofluorescence and biochemical approaches. Typical assays include Western blotting for EHD2/CAV1, RT-qPCR, F-actin staining, and drug sensitivity testing. For further information, contact Ascent Research.
