This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population of the CCDC25 gene in the 786-O human renal cell adenocarcinoma cell line. The polyclonal format provides a heterogenous pool of cells bearing targeted gene disruption, offering a robust loss-of-function model for studying CCDC25 biology without the selective pressure or clonal artifacts associated with single-cell-derived clones. The CRISPR/Cas9-mediated gene inactivation generates a versatile cellular resource suitable for a wide range of functional assays in cancer research.
The 786-O cell line is a well-characterized model of clear cell renal cell carcinoma (ccRCC), originating from a primary renal adenocarcinoma. It harbors a homozygous VHL mutation, which leads to constitutive activation of hypoxia-inducible factor (HIF) pathways and underpins its aggressive, metastatic phenotype. As an epithelial cell line, 786-O retains key features of ccRCC and is widely employed to investigate molecular mechanisms of renal cancer progression and metastasis, making it a relevant host for CCDC25 knockout studies.
CCDC25 functions as a transmembrane receptor for neutrophil extracellular trap (NET) DNA. Upon binding extracellular NET-DNA, CCDC25 activates integrin-linked kinase (ILK) and recruits the adaptor protein ??-parvin. This signaling module drives actin cytoskeleton reorganization, enhancing F-actin polymerization and focal adhesion dynamics to promote cell migration and invasion. Upstream regulators include NETs, extracellular DNA, and inflammatory stimuli that induce NETosis, while downstream effectors encompass ILK, ??-parvin, and components of the actin remodeling machinery. The CCDC25?CILK?C??-parvin axis thus transduces pro-metastatic signals from the tumor microenvironment to the cell motility apparatus.
In the VHL-mutant 786-O background, CCDC25-mediated NET signaling may synergize with HIF-driven programs to amplify metastatic potential. Disrupting CCDC25 in these cells enables researchers to dissect the contribution of NET-induced pathways to renal cancer cell motility and invasion, independent of VHL status. This knockout model is therefore highly valuable for elucidating how tumor?Cmicroenvironment interactions, specifically those involving NETs, facilitate ccRCC dissemination and for identifying CCDC25-dependent vulnerabilities.
Key experimental applications include migration and invasion assays (Boyden chamber, scratch wound), co-immunoprecipitation to assess CCDC25?CILK interaction, western blotting for phospho-ILK, immunofluorescence staining of F-actin, and NET-DNA binding ELISA. The knockout cells are also suitable for flow cytometric validation of surface CCDC25 loss and in vivo metastasis xenograft models to evaluate therapeutic targeting of CCDC25. These tools support investigations into anti-metastatic strategies and the broader role of NETs in cancer. For more information or to place an order, please contact Ascent Research.
