The EHD1 Knockout 786-O Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human 786-O renal cell adenocarcinoma line. This product provides a heterogeneous pool of cells carrying diverse disruptions in the EHD1 gene, establishing a loss-of-function model suitable for studying endocytic recycling without the constraints of clonal selection. The polyclonal format captures the full spectrum of gene-editing outcomes, enabling robust functional analyses of EHD1 deficiency in a cancer cell context.
The 786-O cell line originates from a primary clear cell renal cell carcinoma (ccRCC) and harbors a typical VHL mutation, making it a standard model for renal cancer research. Its epithelial morphology, adherent growth, and well-characterized signaling pathways provide a reliable platform for investigating tumor biology, oncogenic signaling, and therapeutic responses in ccRCC.
EHD1 (Eps15 homology domain-containing protein 1) is an ATPase that functions as a master organizer of endocytic recycling. It acts downstream of Rab11 and Rab35 GTPases, and its activity is modulated by SRC kinase and PIP2 at recycling endosome membranes. EHD1 scaffolds a network of interacting partners, including MICAL-L1, Rabenosyn-5, Syndapin2, SNAP29, and EHD4, to coordinate the return of internalized cargo??such as transferrin receptor (TFRC), integrin beta-1 (ITGB1), and epidermal growth factor receptor (EGFR)??to the plasma membrane. Furthermore, EHD1 couples with cortactin (CTTN) and the Arp2/3 complex to drive actin cytoskeleton remodeling, thereby linking receptor recycling to cell migration and ciliogenesis. Disruption of EHD1 thus derails these interconnected processes, providing a molecular basis for the investigation of trafficking-dependent cellular behaviors.
In the VHL-mutant 786-O background, EHD1 knockout is expected to impair recycling of oncogenic receptors such as EGFR and integrins, potentially attenuating proliferative and migratory signals that drive ccRCC progression. Because EHD1 also participates in primary cilium formation??a structure frequently lost in renal carcinoma??this polyclonal model enables the study of ciliary defects and their contribution to tumorigenesis. The use of a polyclonal population reduces clonal artifacts and better represents the heterogeneous nature of cancer cell populations, offering a physiologically relevant system for dissecting EHD1-dependent phenotypes in kidney cancer.
Researchers can use this knockout pool in diverse assays. Transferrin recycling and EGFR degradation kinetics can be measured, and surface receptor levels quantified by flow cytometry. Western blotting confirms EHD1 loss, while wound healing and Transwell assays assess migration. Cilia formation is visualized by immunofluorescence, and co-immunoprecipitation reveals altered interactomes. RNA-seq and phospho-signaling profiling uncover downstream adaptations. This polyclonal knockout product is ideal for drug target validation, pathway analysis, and screening in ccRCC. For more information, contact Ascent Research.
