The CCDC106 Knockout HeLa Polyclonal Cells provide a CRISPR/Cas9-mediated disruption of the CCDC106 gene within a polyclonal HeLa cell population. CCDC106 is an E3 ubiquitin ligase that catalyzes p53 ubiquitination and proteasomal degradation, thereby repressing p53-dependent transcription. This product offers a versatile platform for investigating ubiquitin-mediated proteolysis and p53 signal transduction in a widely used cervical cancer cell model, without clonal selection bias.
HeLa cells are a human cervical adenocarcinoma line infected with HPV18, which constitutively targets p53 for degradation via the E6 oncoprotein. This background provides a physiologically relevant setting to explore additional p53 regulatory mechanisms, as residual p53 can be activated by DNA damage. HeLa cells?? high proliferation rate and well-characterized nature make them ideal for high-throughput functional studies, and the knockout derivatives enable focused analysis of CCDC106??s role in p53 homeostasis.
CCDC106 directly binds p53 and functions as an E3 ubiquitin ligase, tagging p53 with ubiquitin chains for proteasomal recognition. This action reduces p53 half-life and suppresses transcription of p53 target genes, including the cell cycle inhibitor CDKN1A (p21) and the pro-apoptotic effectors BAX and BBC3/PUMA. CCDC106 expression is itself transcriptionally regulated by p53, forming a negative feedback loop. Cooperating with the major p53 ligase MDM2, CCDC106 fine-tunes p53 abundance in response to cellular stress. Ablation of CCDC106 can thus stabilize p53 and enhance downstream gene expression.
Within HeLa cells, HPV E6 and CCDC106 cooperate to suppress p53; disruption of CCDC106 partially disinhibits p53, even in the presence of E6. This makes the knockout population a powerful model for studying the interplay between viral and cellular p53 regulators. It enables researchers to measure how p53 reactivation alters sensitivity to genotoxic agents like cisplatin, relevant to cervical cancer therapy. The model also helps distinguish E6-dependent from CCDC106-dependent effects on p53 target genes and apoptosis.
Applications include western blotting and RT-qPCR to monitor p53 protein and target gene expression (CDKN1A, BAX, BBC3). Apoptosis and viability assays quantify cell death and proliferation responses. Co-immunoprecipitation and ubiquitination assays define the p53-CCDC106 interaction and ubiquitin modification status. Drug sensitivity screening with chemotherapeutics such as cisplatin or doxorubicin can identify agents that synergize with p53 stabilization. For further details or custom protocols, please contact Ascent Research.
