The EHD4 Knockout 786-O Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal population of the human 786-O renal epithelial cell line, carrying a targeted disruption of the EHD4 gene. This heterogeneous knockout pool offers a powerful loss-of-function model for studying endocytic recycling and receptor trafficking in the context of clear cell renal cell carcinoma (ccRCC). The polyclonal format captures a range of knockout-induced phenotypes without clonal selection bias, enabling robust functional genomics investigations. Created via CRISPR/Cas9-mediated gene disruption, this product is intended for advanced applications in cancer biology, signal transduction, and drug discovery.
The 786-O cell line originates from a primary clear cell renal adenocarcinoma and is deficient in the von Hippel-Lindau (VHL) tumor suppressor, resulting in constitutive stabilization of hypoxia-inducible factor 2?? (HIF-2??). This VHL-deficient background drives a pseudohypoxic state with altered gene expression, enhanced growth factor signaling, and increased metastatic potential. As a well-validated model for ccRCC, 786-O cells are instrumental for elucidating oncogenic mechanisms and testing therapeutic interventions that target the HIF pathway or downstream signaling modules.
EHD4 (Eps15 homology domain-containing protein 4) is a critical regulator of endocytic recycling, orchestrating the return of internalized receptors, particularly EGFR, from endosomes to the plasma membrane. EHD4 interacts with key endosomal factors, including Rab11, Rab5, clathrin, AP2, EHBP1, and ARF6, to control the surface availability and signaling lifespan of receptors. By sustaining EGFR levels at the cell surface, EHD4 promotes activation of the MAPK and AKT pathways, which govern proliferation and survival. Additionally, EHD4 modulates integrin recycling, influencing focal adhesion turnover, cell migration, and invasion??processes directly relevant to cancer metastasis.
In 786-O cells, EHD4 knockout is anticipated to disrupt EGFR and integrin recycling, attenuating AKT and ERK signaling, and impairing migratory and invasive behaviors. Given the pivotal role of these pathways in ccRCC progression and metastasis, the EHD4 knockout model provides a clinically relevant tool to dissect the interplay between membrane trafficking and tumor aggressiveness. It also facilitates the identification of synthetic vulnerabilities arising from combined VHL deficiency and EHD4 loss.
Researchers can utilize this polyclonal knockout pool in an array of assays: EGFR recycling kinetics, immunofluorescence for endosomal markers, western blotting for phospho-EGFR/AKT/ERK, and functional migration/invasion tests. Xenograft studies enable in vivo assessment of metastatic potential and therapeutic response. The model is also suited for drug screens targeting endocytic machinery or receptor tyrosine kinase signaling. For additional information and technical support, please contact Ascent Research.
