The DNAJB4 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 human hepatocellular carcinoma line. This product provides a heterogeneous knockout pool for loss-of-function studies of DNAJB4, a co-chaperone implicated in protein folding, endocytosis, and tumor suppression. The polyclonal format offers a convenient model to assess DNAJB4-dependent phenotypes without the need for clonal isolation, capturing a range of editing outcomes.
Huh-7 is an epithelial cell line established from a well-differentiated hepatocellular carcinoma of a 57-year-old Japanese male with hepatitis C virus infection. These adherent cells maintain hepatic characteristics and are widely employed in liver cancer research, including studies on viral oncogenesis, drug metabolism, and tumor suppressor pathways. The cell line’s well-characterized molecular landscape makes it a robust host for gene perturbation studies, and its tumorigenic background provides a disease-relevant context for investigating DNAJB4’s role in hepatocellular carcinoma.
DNAJB4 acts as a J-domain co-chaperone for Hsc70 (HSPA8), facilitating protein folding and clathrin-mediated endocytosis. Its expression is regulated by HSF1, ATF6, and XBP1, and is frequently silenced by promoter hypermethylation in cancers. DNAJB4 interacts with clathrin, p53, and IKK??, and its loss leads to activation of STAT3, ??-catenin, ERK1/2, and JNK signaling. Consequently, DNAJB4 deficiency drives hepatocellular carcinoma progression by unleashing MAPK/ERK and Wnt/??-catenin pathway activity, thereby promoting tumor cell proliferation, survival, and invasion.
In Huh-7 cells, which exhibit basal activation of MAPK and Wnt cascades, DNAJB4 knockout provides a model system to study the transition from chaperone-mediated tumor suppression to malignancy. The polyclonal population allows examination of phenotypic variability and adaptive responses that may arise after gene disruption. This model is particularly suited for probing the crosstalk between DNAJB4, STAT3, and ??-catenin in liver cancer.
Applications include investigation of chaperone-dependent tumor suppression, clathrin-mediated endocytosis, and signal transduction in hepatocellular carcinoma. Representative assays include Western blotting, RT-qPCR, immunofluorescence, flow cytometry, endocytosis assays, cell proliferation and migration/invasion studies, phospho-STAT3 analysis, ??-catenin reporter assays, and drug sensitivity screens. This knockout product supports functional genomics, target validation, and preclinical drug discovery. For more information, contact Ascent Research.
