This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from Huh-7 cells, in which the DNAJA2 gene has been disrupted to create a loss-of-function model. The polyclonal knockout format provides a heterogeneous pool of edited cells, enabling robust analysis of DNAJA2-dependent processes without the limitations of clonal selection. This model is designed for investigating co-chaperone functions in proteostasis, stress signaling, and disease contexts.
Huh-7 is a well-differentiated human hepatocellular carcinoma cell line that retains key hepatocyte features, including liver-specific metabolism and permissiveness to hepatitis B and C viruses. As a model for hepatic biology and cancer, Huh-7 cells provide a physiologically relevant background for studying the roles of chaperones like DNAJA2 in protein quality control, viral replication, and oncogenic signaling. This host cell line is thus an ideal platform for dissecting how disruptions in proteostasis influence liver tumor cell behavior.
DNAJA2 functions as a critical co-chaperone, binding misfolded proteins and delivering them to HSP70 family members (e.g., HSPA1A/HSPA8), where it stimulates ATPase activity to drive conformational changes that direct clients toward refolding or ubiquitin-mediated proteasomal degradation. This activity is regulated upstream by HSF1 transcriptional activation during heat stress, as well as by the unfolded protein response and oxidative stress. DNAJA2 interacts with a network of partners including HSP90B1, BAG family co-chaperones, STIP1, and the E3 ligase CHIP/STUB1, which together determine client fate. The canonical pathway??HSF1 ?? DNAJA2 ?? HSP70 ?? client protein refolding or degradation??highlights its central role in maintaining cellular proteostasis and mitigating proteotoxic stress.
In hepatocellular carcinoma, proteostatic dysregulation is often exploited for survival and drug resistance. This knockout model enables dissection of how loss of DNAJA2 affects cancer cell viability, heat shock responses, and the replication of oncogenic viruses such as HBV and HCV. The relevance extends beyond liver cancer to neurodegenerative diseases marked by protein aggregation and to cystic fibrosis, where chaperone-assisted folding of CFTR is crucial. By disrupting DNAJA2, researchers can probe the dependency of tumor cells on HSP70-driven quality control and identify potential therapeutic vulnerabilities.
Representative applications include HSF1 luciferase reporter assays, RT-qPCR and western blotting for heat shock genes, immunofluorescence for protein aggregates, co-immunoprecipitation of DNAJA2?CHSP70 complexes, and flow cytometry for apoptosis. These cells support drug screens targeting chaperones and mechanistic studies of host?Cvirus interactions. For more information, contact Ascent Research.
