The CCDC85C Knockout MES-OV Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population in the human ovarian clear cell carcinoma cell line MES-OV, with targeted disruption of the CCDC85C gene (Homo sapiens). This polyclonal knockout model comprises a heterogeneous pool of edited cells, collectively representing a loss-of-function state for CCDC85C, a centrosomal protein that negatively regulates centriole duplication and promotes primary cilium formation. The polyclonal format captures the diversity of CRISPR/Cas9-mediated mutations across the cell population, providing a robust tool for functional studies without the clonal selection bias inherent to single-cell-derived lines.
The MES-OV cell line is an ovarian surface epithelial cell line derived from the malignant ascites of a patient with ovarian clear cell carcinoma, a histologic subtype associated with poor prognosis and chemoresistance. These cells retain tumorigenic properties and serve as a clinically relevant model for investigating ovarian cancer biology, particularly centrosome abnormalities and ciliary signaling defects that contribute to genomic instability and tumor progression.
CCDC85C localizes to the centrosome and acts as a restrictive factor in centriole biogenesis by modulating the assembly of key duplication components, including PLK4, SAS-6, STIL, and CPAP, and it also facilitates primary cilium formation through interactions with ciliogenic machinery such as IFT88 and ARL13B. The protein is regulated by FOXM1 and E2F transcription factors and physically interacts with centrosomal scaffold proteins CEP120, CEP135, CDK5RAP2, and pericentrin. Its knockout disrupts these interactions, leading to unchecked centriole overduplication, centrosome amplification, and impaired ciliogenesis, which can alter cell cycle progression and attenuate Hedgehog signaling pathways that depend on the primary cilium.
In the context of MES-OV cells, disruption of CCDC85C creates a relevant model to study how centrosome amplification and defective primary cilia contribute to ovarian cancer pathogenesis. Centrosome aberrations are common in ovarian clear cell carcinoma and are linked to mitotic errors, aneuploidy, and aberrant signaling. This polyclonal knockout pool enables researchers to examine the phenotypic consequences of CCDC85C loss across a diverse genetic background, reflecting the heterogeneity of tumor cell populations. The model is particularly suited for delineating the molecular interplay between centrosome duplication machinery, ciliary assembly factors, and downstream effectors such as the Hedgehog pathway components, and for testing therapeutic agents that target centrosome clustering or restore ciliary function.
Typical applications include centrosome duplication studies using immunofluorescence for ??-tubulin to enumerate centrioles, cell cycle analysis via flow cytometry for DNA content, assessment of primary cilium formation by acetylated tubulin immunostaining, and evaluation of Hedgehog signaling through RT-qPCR for target genes like GLI1 and PTCH1. Further functional assays encompass western blotting for CCDC85C and interacting partners, cell proliferation assays, and mitotic index quantification to gauge genomic instability. The polyclonal knockout format is well-suited for drug screening campaigns aiming to identify compounds that selectively kill cells with supernumerary centrosomes or restore ciliogenesis. For additional information on validation data and culture conditions, please contact Ascent Research.
