The CCDC91 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of Huh-7 cells with targeted disruption of the CCDC91 gene, designed to ablate CCDC91 protein expression. This knockout model provides a powerful tool for investigating Golgi apparatus organization and intracellular trafficking in a hepatocellular carcinoma background. The polyclonal format offers a heterogeneous knockout pool, minimizing clonal selection artifacts and enabling population-level functional analyses.
Huh-7 is an epithelial-like adherent cell line derived from a liver tumor of a 57-year-old Japanese male, widely utilized as a model for hepatocellular carcinoma and hepatocyte biology. These cells express characteristic hepatic markers such as albumin and alpha-fetoprotein, and retain many metabolic and secretory properties of primary hepatocytes, making them highly suitable for studies of liver-specific pathology, including viral hepatitis, cirrhosis, and malignant transformation.
CCDC91 encodes a coiled-coil domain-containing protein that localizes to the Golgi apparatus and functions as a scaffold critical for maintaining cisternal stack organization and facilitating vesicle-mediated transport. It directly interacts with key Golgi structural proteins GOLGA2 (GM130) and GOLGB1, the COG tethering complex, and the small GTPase RAB1A, operating within the COPI/COPII trafficking machinery. Upstream, CCDC91 is regulated by the hepatocyte-enriched transcription factors HNF4A and CEBPA, as well as by receptor tyrosine kinase signaling through EGFR and MET. Knockout of CCDC91 disrupts protein secretion and receptor trafficking, resulting in aberrant glycosylation and impaired presentation of cell surface receptors.
In the Huh-7 hepatocellular carcinoma background, loss of CCDC91 is expected to severely dysregulate the secretory pathway and membrane protein trafficking, processes that are frequently co-opted by cancer cells to sustain proliferation, invasion, and immune evasion. This knockout model enables detailed dissection of how Golgi-dependent secretion impacts hepatocellular carcinoma progression, including effects on growth factor receptor presentation, extracellular matrix remodeling, and cell?Ccell communication. Moreover, since Huh-7 cells secrete albumin, this system provides a quantifiable functional readout of secretory pathway integrity in a disease-relevant context.
Key experimental applications include immunofluorescence microscopy and Western blotting for Golgi markers such as GM130 and TGN46, ELISA-based quantification of albumin secretion, and mass spectrometry-based secretome profiling to assess global protein secretion changes. The model is also well-suited for cell proliferation (MTT or BrdU), migration and invasion (wound healing or Transwell), and cell cycle analysis by flow cytometry, facilitating liver cancer drug screening. Additionally, RT-qPCR can be employed to monitor hepatic gene expression changes. For technical inquiries or custom applications, please contact Ascent Research.
