The CCDC97 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated in the 786-O renal cell carcinoma cell line. This product provides a loss-of-function model for the CCDC97 gene, enabling investigation of its role in centrosome function and cell cycle regulation. The polyclonal population offers a heterogeneous knockout background suitable for pooled analysis and functional screening in a disease-relevant cellular context.
This model employs the 786-O human clear cell renal cell carcinoma (ccRCC) cell line, which harbors a VHL mutation leading to constitutive stabilization of hypoxia-inducible factors (HIFs) and persistent angiogenic signaling. 786-O cells are widely used to study VHL-mediated tumorigenesis, the HIF pathway, and therapeutic interventions targeting downstream oncogenic programs. The combination of VHL deficiency and CCDC97 disruption provides a unique platform to examine cross-talk between hypoxia signaling and cell division machinery.
CCDC97 localizes to centrosomes and is essential for proper centriole duplication and mitotic spindle assembly. Mechanistically, CCDC97 is transcriptionally regulated by TP53, FOXM1, and E2F1, and it interacts directly with centrosomal and cell-cycle regulators including PLK1, AURKA, CEP152, and TUBG1. Upon knockout, centrosome amplification occurs, causing chromosomal instability and triggering the ATM-CHEK1 DNA damage response. This activates p53 signaling, leading to transcription of downstream targets CDKN1A (p21) and BAX, which mediate cell cycle arrest and apoptosis. Key pathway components affected include cyclins A2 and B1 (CCNA2, CCNB1), the mitotic phosphatase CDC25C, and centrosome duplication factors PLK4, STIL, and SAS6.
In the context of 786-O cells, CCDC97 loss exacerbates inherent genomic instability resulting from VHL-dependent dysregulation. The interplay between HIF-driven oncogenic programs and CCDC97-mediated centrosome control offers a sophisticated model to dissect tumor-suppressive mechanisms in ccRCC. This system enables evaluation of how centrosome dysfunction contributes to chromosomal instability and mitotic catastrophe in a kidney cancer background, and whether p53-dependent apoptotic barriers are activated or bypassed during tumor evolution.
This polyclonal knockout cell population is suited for a broad range of research applications. Centrosome biology and mitosis can be examined using immunofluorescence staining for centrosome markers and tubulin, while cell cycle perturbations are quantifiable via flow cytometry. Western blotting of cyclins, CDKs, and apoptotic markers, along with Annexin V apoptosis assays and MTS/EdU proliferation assays, allows mechanistic validation. Transcriptomic profiling by RNA-seq can reveal global gene expression changes. The model also serves for screening small-molecule mitotic kinase inhibitors targeting PLK1 or AURKA. For additional technical details, please contact Ascent Research.
