The CCDC50 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, designed for loss-of-function studies of the CCDC50 gene. This product harnesses heterogeneous gene disruption across the cell pool, providing a robust system for analyzing CCDC50-dependent pathways without the need for clonal isolation. The polyclonal format is particularly suited for bulk biochemical assays and population-level investigations where diverse editing events more closely mimic the complexity of primary tissue responses.
HeLa cells serve as the host background, representing a widely utilized human cervical adenocarcinoma epithelial line positive for HPV18. As an immortalized cell model with robust proliferation and transformed characteristics, HeLa is a cornerstone for cancer biology, signal transduction research, and drug discovery. Their epithelial origin and stable growth make them an optimal platform for studying the interplay between autophagy and inflammatory signaling in a malignancy-relevant context.
CCDC50 encodes a ubiquitin-binding adaptor protein that negatively regulates NF-??B signaling by selectively targeting NEMO (IKBKG) for autophagic degradation. Knockout of CCDC50 disrupts this control mechanism, leading to NEMO accumulation, sustained IKK complex activation, and downstream phosphorylation and degradation of I??B??, followed by nuclear translocation of RELA/p65. This results in constitutive transcription of pro-inflammatory cytokines. The protein also interacts with key regulators including SQSTM1/p62, TRAF6, and RIPK1, and is modulated by upstream signals such as TNF-??, IL-1??, LPS, and oxidative stress, placing it at a critical node between selective autophagy and immune signaling.
In the HeLa cervical cancer environment, CCDC50 loss establishes a state of chronic NF-??B activation and impaired autophagy flux, partly mimicking inflammatory oncogenic processes. This model is invaluable for dissecting how ubiquitin-dependent clearance of NEMO restrains inflammatory gene expression, and for exploring potential vulnerabilities in HPV-associated malignancies where oncoproteins may already perturb cellular homeostasis. The system also facilitates studies on the broader role of autophagy in cancer cell survival and therapy resistance.
Research applications include NF-??B luciferase reporter assays, western blotting of phosphorylated I??B?? and p65, RT-qPCR of downstream targets, LC3 puncta immunofluorescence for autophagy assessment, and multiplex cytokine ELISA. The model further supports hearing loss research due to CCDC50??s link to autosomal dominant nonsyndromic hearing loss, and can be employed in compound screens targeting ubiquitin-mediated autophagy or NF-??B modulation. For additional technical details, please contact Ascent Research.
