The CCDC9B Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa human cervical adenocarcinoma cell line, engineered for in vitro investigation of the coiled-coil domain-containing protein 9B (CCDC9B) loss-of-function phenotype. This polyclonal format introduces a spectrum of gene disruptions across a heterogeneous pool of edited cells, enabling robust functional genomic screening without the constraints of single-cell cloning. The population-level representation of CCDC9B deficiency provides a versatile model for studying gene function in pooled assay formats.
HeLa cells, an HPV18-positive cervical adenocarcinoma line, are immortalized by the E6 and E7 oncoproteins, which inactivate p53 and Rb, respectively, thereby abrogating G1/S checkpoints. Their epithelial nature and intact centrosome duplication machinery make them a suitable model for investigating mitotic regulation. The rapid doubling time and transfectability of HeLa cells enable high-throughput imaging and biochemical assays.
CCDC9B encodes a coiled-coil protein predicted to participate in centrosome duplication and microtubule organization, thereby influencing cell cycle progression. It is likely regulated by CDK1/cyclin B and PLK4 signaling, central to mitotic entry and centriole biogenesis. CCDC9B interacts with centrosomal scaffold and microtubule-associated proteins, recruiting components such as CEP152, Centrin, SAS-6, and STIL. Acting downstream of PLK4, it may promote procentriole formation and spindle assembly; its disruption thus impairs these processes and induces mitotic defects.
In HeLa cells, where HPV-mediated genomic instability often causes centrosome amplification, CCDC9B knockout can reveal how cancer cells cope with such aberrations. The polyclonal model maintains HeLa heterogeneity while disrupting CCDC9B, enabling study of selective pressures and compensatory pathways. Loss of CCDC9B function may exacerbate mitotic errors, affecting proliferation, apoptosis, or cell fate, providing a system to dissect viral transformation effects on centrosome integrity.
Key applications include immunofluorescence staining of centrosomal markers, flow cytometric cell cycle profiling, and Western blotting for mitotic regulators such as phospho-histone H3. The polyclonal format supports functional genomics screens, RT-qPCR, and proliferation/apoptosis assays. Researchers can use this model to explore centrosome biology in HPV-driven cancers, screen for synthetic lethality, or test PLK4/CDK1 inhibitors. For further details, contact Ascent Research.
