The CCDC85C knockout 786-O polyclonal cells consist of a CRISPR/Cas9-edited population of human renal cell carcinoma cells, specifically designed for loss-of-function studies of the CCDC85C gene. As a polyclonal knockout model, this product provides a heterogeneous cell pool with gene disruptions, bypassing clonal selection biases and enabling robust population-level phenotype analysis. The CRISPR/Cas9 system has been used to introduce gene disruption across multiple cells, ensuring effective target-gene knockout while preserving the parental line??s background. This format is ideal for large-scale functional genomics screens and rapid characterization of gene function without the time and artifacts associated with clonal isolation.
The 786-O cell line is a widely utilized model of human clear cell renal cell adenocarcinoma, originally derived from a primary renal tumor. These cells harbor a characteristic VHL mutation, resulting in constitutive HIF pathway activation and mimicking key aspects of renal cancer biology. 786-O cells grow adherently and are amenable to genetic manipulation, making them a preferred platform for CRISPR-based knockout experiments. Their well-documented tumorigenic properties, including dysregulated proliferation and migration, provide a physiologically relevant context for investigating genes implicated in centrosome and cilia function, such as CCDC85C.
CCDC85C encodes a coiled-coil domain-containing protein with a predicted scaffolding role at the centrosome and primary cilium. Although its full interactome remains undefined, CCDC85C has been shown to interact with DISC1, a key centrosomal and ciliary protein involved in psychiatric disorders. This interaction suggests involvement in centrosome organization and ciliogenesis pathways, where it may facilitate microtubule anchoring, centriole stability, or assembly of signaling complexes. Consequently, CCDC85C loss could impair centrosome duplication, ciliogenesis, and cilia-dependent signaling, potentially impacting cell cycle progression, polarity, and migration.
In the renal cell carcinoma context of 786-O cells, CCDC85C knockout offers a valuable tool to examine how centrosomal and ciliary disruptions impact cancer phenotypes. Renal cancers frequently display centrosome amplification and ciliary signaling defects that contribute to tumorigenesis. By eliminating CCDC85C, researchers can dissect its specific contribution to these processes, such as centrosome overduplication, cilia-mediated Hedgehog or Wnt signaling modulation, and subsequent effects on proliferation, invasion, and drug sensitivity. This model is particularly relevant for identifying centrosome-related dependencies in renal cell carcinoma that could be exploited therapeutically.
This polyclonal knockout model is suited for functional characterization of CCDC85C using immunofluorescence centrosome staining, western blotting for protein analysis, and RT-qPCR for transcript quantification. Co-immunoprecipitation experiments enable examination of CCDC85C-DISC1 complexes. Cell proliferation and migration assays provide quantitative readouts of cancer-related phenotypes. Additionally, the cells can be used in high-content imaging screens for centrosome or cilia abnormalities and in RNA-seq studies to uncover transcriptional changes. For technical assistance, protocol inquiries, or customization, please contact Ascent Research.
