This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 cell line, in which the EHD3 gene has been disrupted to generate a loss-of-function model. The polyclonal pool offers a heterogeneous collection of cells with targeted gene disruption, enabling robust analysis of EHD3-dependent endocytic trafficking without clonal artifacts. As a research tool, these polyclonal knockout cells are ideal for dissecting the role of EHD3 in membrane dynamics and receptor recycling in a hepatocellular carcinoma context.
Huh-7 is a human hepatocellular carcinoma cell line established from a well-differentiated liver tumor of a 57-year-old male, and it is negative for hepatitis C virus. This cell line serves as a widely used model for hepatocellular carcinoma and liver biology, retaining key features of hepatic epithelial cells and oncogenic signaling pathways. Its stable growth characteristics and genetic manipulability make Huh-7 an excellent host for loss-of-function studies targeting genes involved in endocytic trafficking and cancer progression.
EHD3 functions as an endocytic recycling protein that facilitates the return of internalized receptors, such as EGFR and transferrin receptor, from early endosomes to the plasma membrane. It operates through interactions with Rab11, Rab8, Arf6, syndapin, and actin filaments to coordinate membrane fission and vesicle transport. EHD3 is activated downstream of growth factor stimulation (e.g., EGF) and androgen receptor signaling, and it regulates EGFR recycling to the plasma membrane, transferrin receptor trafficking, and processes of cell migration and invasion. Disruption of EHD3 by CRISPR/Cas9 perturbs this recycling network, leading to altered receptor signaling dynamics and impaired cellular motility.
In the context of hepatocellular carcinoma, EHD3 expression is frequently altered, and its knockout in Huh-7 cells provides a relevant model to investigate the molecular underpinnings of cancer progression. EHD3-dependent endocytic recycling has been implicated in sustaining oncogenic signaling through receptor tyrosine kinases, and its loss may affect tumor cell migration and invasion. Consequently, this polyclonal knockout model is valuable for exploring how endosomal trafficking influences hepatocellular carcinoma pathogenesis and for identifying potential therapeutic vulnerabilities.
These polyclonal EHD3 knockout Huh-7 cells are applicable to a range of experimental techniques, including western blotting to confirm protein loss, transferrin uptake assays and immunofluorescence to visualize endocytic defects, migration and invasion assays to assess metastatic potential, co-immunoprecipitation to probe protein interactions, and RNA-seq for transcriptomic analysis. They support studies of EGFR signaling, drug resistance mechanisms, and clathrin-independent endocytosis. For detailed technical specifications or ordering information, please contact Ascent Research.
