The EHBP1 Knockout 786-O Polyclonal Cells are a polyclonal population of 786-O human renal cell carcinoma cells in which CRISPR/Cas9 has been used to disrupt the EHBP1 gene, creating a heterogeneous knockout model. The polyclonal format captures a mixture of gene-edited cells without clonal selection, offering a robust system for functional studies that reflect population-level variation.
The parental 786-O line is an established epithelial model of clear cell renal cell carcinoma (ccRCC), originally derived from a primary tumor. It harbors a known mutation in the VHL tumor suppressor gene, resulting in constitutive activation of hypoxia-inducible factor signaling, a hallmark of ccRCC. This genetic background is widely utilized for investigating renal cancer biology, including pathways governing cell adhesion, migration, and endocytosis.
EHBP1 encodes a scaffolding protein that physically links EH domain-containing proteins EHD1 and EHD2 to the actin cytoskeleton through interactions with cortactin and non-muscle myosin heavy chain IIA (NMHC-IIA). This bridging function is essential for endosomal tubulation and recycling of internalized cargo, such as integrins and growth factor receptors. Upstream signals from integrin engagement and growth factors activate EHBP1, recruiting it to endosomes where it coordinates EHD family proteins and the actin polymerization machinery to drive vesicle fission and transport. Consequently, EHBP1 serves as a key regulator of endocytic recycling and actin-dependent processes like cell adhesion and migration.
In the VHL-mutant 786-O ccRCC context, loss of EHBP1 is predicted to further impair endosomal trafficking and cytoskeletal organization, exacerbating defects in cell?Cmatrix adhesion and directional migration. This makes the knockout model particularly valuable for studying the interplay between VHL loss and EHBP1 function in driving the invasive phenotype of renal carcinoma. It allows researchers to probe whether EHBP1 functions as a tumor suppressor or dependency in ccRCC progression.
This polyclonal knockout cell population is ideal for a wide range of experimental applications, including functional characterization of EHBP1 in renal cell carcinoma, endocytic recycling assays such as transferrin uptake and recycling, wound healing migration assays, and co-immunoprecipitation studies of EHBP1 interactions with EHD proteins and cortactin. Western blotting and flow cytometry can confirm EHBP1 disruption and assess downstream effects on cytoskeletal markers. The mixed genetic background complements monoclonal lines by enabling analysis of population-level phenotypes. For further information, please contact Ascent Research.
