The CCDC124 Knockout HeLa Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, targeting the CCDC124 gene of Homo sapiens. This loss-of-function model disrupts CCDC124 through CRISPR-mediated gene editing, generating a heterogeneous pool of knockout variants without single-cell cloning. Such polyclonal populations are ideal for aggregate functional studies, offering a robust system to assess gene function in a human epithelial carcinoma background. The product serves as a versatile tool for investigating cytokinesis and abscission mechanisms.
HeLa cells originate from an HPV18-positive cervical adenocarcinoma and display epithelial characteristics. Extensively employed in cell biology, drug discovery, and cancer research, this line provides a reproducible platform for dissecting molecular pathways. Integrating a CCDC124 knockout into this established model enables direct examination of gene function within a well-characterized transformed environment, particularly relevant for cytokinesis studies given HeLa cells’ rapid proliferation and mitotic activity.
CCDC124 encodes a key midbody protein that facilitates the final abscission step of cytokinesis. It is recruited to the midbody via interaction with CEP55, which scaffolds the assembly of ESCRT-III components??notably CHMP4B??and the ATPase VPS4, alongside adaptors ALIX and TSG101. This machinery severs the intercellular membrane, assisted by the microtubule-severing enzyme spastin. CCDC124 acts downstream of mitotic kinases, including PLK1, ensuring abscission timing is coupled to cell cycle progression. Disruption of CCDC124 impedes ESCRT-III recruitment, blocking membrane scission, resulting in multinucleated cells. Such cytokinesis failure contributes to genomic instability and can promote tumorigenesis.
Within the HeLa cervical adenocarcinoma context, CCDC124 knockout generates multinucleated cells, mimicking defects seen in certain human cancers. This phenotype, combined with the existing HPV18-induced genomic instability of HeLa cells, amplifies division errors and provides a tractable system for studying polyploidization and aneuploidy. The model is invaluable for exploring how tumor cells respond to cytokinesis failure and the mechanisms that normally suppress tetraploidization. Notably, with PLK1 frequently overexpressed in malignancies, the CCDC124 pathway represents a candidate vulnerability in cancer.
The CCDC124 knockout polyclonal population supports diverse assays: time-lapse microscopy captures real-time abscission failure, while immunofluorescence localizes midbody proteins like CEP55 and CHMP4B. Western blot and RT-qPCR verify CCDC124 ablation, and flow cytometry reveals DNA content shifts indicative of multinucleation. Quantitative abscission and multinucleation assays enable phenotypic measurement, facilitating functional genomics screening and evaluation of small molecules targeting cytokinesis. For additional product information, please contact Ascent Research.
