The EHD3 Knockout 786-O Polyclonal Cells are a human polyclonal cell population derived from the 786-O clear cell renal cell carcinoma line, engineered via CRISPR/Cas9-mediated gene disruption of the EHD3 gene. This polyclonal knockout cell pool provides a heterogeneous loss-of-function model to study EHD3-dependent processes. No single-cell cloning was performed, maintaining genetic diversity while disrupting target protein expression.
786-O is a widely used human renal cell carcinoma cell line isolated from a primary clear cell adenocarcinoma. These cells harbor a VHL tumor suppressor mutation, characteristic of clear cell renal cell carcinoma, and exhibit epithelial morphology. They serve as a standard in vitro system for investigating kidney cancer biology, including hypoxia response, angiogenesis, and metastatic mechanisms.
EHD3 belongs to the Eps15 homology (EH) domain-containing protein family, functioning primarily in endocytic recycling and receptor trafficking. It interacts with EHD1, EHD2, Rab GTPases (such as Rab11), actin, and membrane curvature proteins to regulate the recycling of cargoes like transferrin receptor and integrins back to the plasma membrane. EHD3 activity is modulated by upstream growth factor and integrin signaling pathways, and it controls downstream receptor recycling and cell migration machinery. By regulating the surface levels of adhesion and signaling receptors, EHD3 influences cellular responses to extracellular cues.
In the 786-O carcinoma context, EHD3 knockout likely disrupts efficient receptor recycling, impairing cell adhesion, migration, and signal transduction. Given the role of integrin recycling in cancer cell motility and metastasis, this model allows investigation of how defective endocytic trafficking affects renal cell carcinoma progression. The VHL-mutant background further provides a clinically relevant genetic environment to study EHD3 in a tumor context.
This knockout model is suited for assessing the impact of EHD3 loss on endocytic trafficking dynamics through transferrin recycling assays and immunofluorescence for endosomal markers. It enables analysis of cell migration and invasion using wound healing or Transwell assays, and evaluation of proliferation effects. Additionally, it can be employed in drug discovery screens targeting renal cell carcinoma pathways dependent on receptor trafficking. Scientists can use this polyclonal knockout population for western blotting to confirm EHD3 disruption and examine downstream signaling nodes. For further information or to request a quote, please contact Ascent Research.
