The DNAJC3 Knockout Huh-7 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DNAJC3 gene in the Huh-7 human hepatocellular carcinoma cell line. This polyclonal pool comprises a heterogeneous mixture of cells carrying diverse CRISPR-mediated edits at the target locus, creating a functional loss-of-function model for investigating the roles of DNAJC3 (p58IPK) in endoplasmic reticulum (ER) stress signaling, translational control, and cancer biology. The product is optimized for researchers requiring a robust, scalable system to study DNAJC3-dependent pathways without the constraints of single-cell-derived clones.
Huh-7 cells, the host model, are epithelial cells derived from a well-differentiated hepatocellular carcinoma of a Japanese male. These cells retain key hepatocyte features, including expression of liver-specific enzymes and susceptibility to hepatitis C virus infection, making them a widely employed system in liver cancer research, drug metabolism, and viral pathogenesis studies. The adherent growth and stable karyotype of Huh-7 cells facilitate reproducible transfection, lentiviral transduction, and functional assays, providing a reliable background for gene disruption experiments.
DNAJC3 encodes p58IPK, an ER lumenal co-chaperone that functions as a critical negative regulator of the unfolded protein response (UPR). Mechanistically, DNAJC3 interacts with HSPA5 (BiP) and directly binds the ER stress kinases PERK (EIF2AK3) and PKR (EIF2AK2), attenuating their kinase activity. This inhibition reduces phosphorylation of the translation initiation factor eIF2??, allowing translational recovery after stress resolution. Consequently, DNAJC3 suppresses the downstream effectors ATF4 and CHOP (DDIT3) and modulates XBP1 splicing via IRE1??. In the absence of DNAJC3, PERK/PKR signaling is enhanced, leading to sustained eIF2?? phosphorylation, elevated ATF4 translation, and heightened sensitivity to ER stress-induced apoptosis. DNAJC3 also interacts with DNAJB11, placing it at a hub of the integrated stress response (ISR).
In the context of Huh-7 hepatocellular carcinoma cells, DNAJC3 knockout amplifies UPR signaling, offering a physiologically relevant model for investigating ER stress in liver cancer. Hepatocytes are inherently susceptible to ER stress due to high secretory protein load, and Huh-7 cells recapitulate this vulnerability. The polyclonal nature of this knockout population mirrors the heterogeneity found in tumor environments, enabling studies of signaling thresholds, compensatory mechanisms, and drug sensitivity variations. This model is particularly useful for dissecting PERK-dependent chemoresistance, exploring viral co-option of UPR pathways, and testing pharmacological modulators of the ISR.
The DNAJC3 Knockout Huh-7 Polyclonal Cells support a broad range of experimental applications, including western blotting for UPR markers (p-eIF2??, ATF4, CHOP), RT-qPCR for target gene expression, immunofluorescence detection of ER stress proteins, and apoptosis or viability assays under treatment with ER stress inducers such as thapsigargin or tunicamycin. Researchers can use this system for drug screening against the PERK/eIF2??/ATF4 axis, metabolic disease modeling, and studies on translational regulation in hepatocellular carcinoma. The polyclonal design provides robust flexibility for investigating DNAJC3 biology across diverse stress conditions. For further information, please contact Ascent Research.
