The CCDC91 Knockout HeLa Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed for the study of CCDC91 function in human cells. Derived through CRISPR/Cas9-mediated gene disruption, this polyclonal pool retains a heterogeneous mixture of edited alleles, providing a robust loss-of-function model without clonal isolation. The knockout product format is ideally suited for applications requiring population-level analyses of CCDC91-dependent processes, including protein sorting, endosomal trafficking, and lysosomal biology.
The host cell line, HeLa, is an epithelial cell line originating from a cervical adenocarcinoma and is positive for human papillomavirus 18 (HPV-18). HeLa cells are among the most extensively utilized human cell lines in biomedical research, offering well-characterized growth kinetics, genetic tractability, and a conserved secretory and endocytic machinery. Their cancerous origin and immortalized nature make them a relevant model for investigating oncogenic mechanisms and intracellular trafficking pathways that may be dysregulated in cancer.
CCDC91 encodes a coiled-coil domain-containing protein that functions as an accessory factor for the AP-1 adaptor complex, critical for clathrin-mediated vesicle transport from the trans-Golgi network (TGN) to endosomes. CCDC91 interacts directly with AP-1 subunits (AP1G1, AP1B1, AP1S1), clathrin triskelion, and GGA proteins, facilitating cargo sorting and vesicle formation. Disruption of CCDC91 impairs the trafficking of lysosomal enzymes, such as cathepsin D, via the mannose-6-phosphate receptor pathway, leading to mis-sorting, potential enzyme secretion, and lysosomal dysfunction. Downstream consequences may affect the degradation of cell surface receptors and extracellular matrix components, linking CCDC91 to broader cellular homeostasis.
In the HeLa cell context, CCDC91 knockout provides a powerful tool to dissect the molecular mechanisms underlying cargo sorting at the TGN and its impact on cancer cell behavior. Given the role of lysosomal exocytosis and protease secretion in tumor invasion and metastasis, this model allows investigation of how disrupted endolysosomal trafficking influences the aggressive properties of cervical cancer cells. It also serves as a cellular platform for studying lysosomal storage-like disorders, where aberrant hydrolase trafficking mimics certain disease phenotypes.
Researchers can employ this knockout model in a variety of assays, including immunofluorescence microscopy to visualize TGN and endosomal markers, western blotting to detect lysosomal enzyme secretion, transferrin uptake assays to assess endocytic efficiency, flow cytometry for surface LAMP1 levels, and co-immunoprecipitation to examine AP-1 complex integrity. These applications support studies in cancer cell invasion, lysosomal storage disease modeling, and endosomal drug delivery. For additional details, please contact Ascent Research.
