The DNAJA1 Knockout SK-HEP-1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which targeted disruption of the DNAJA1 gene abrogates its expression. This genetically mixed pool, originating from the SK-HEP-1 host cell line, provides a loss-of-function model that retains cellular heterogeneity, minimizing biases associated with single-cell-derived clones. It is designed for systematic investigation of DNAJA1??s roles in protein homeostasis and oncogenic signaling.
SK-HEP-1 is an ascites-derived human hepatic adenocarcinoma cell line extensively employed as a model for hepatocellular carcinoma (HCC). These cells exhibit an epithelial morphology and retain hallmark features of liver cancer, including rapid proliferation, migratory behavior, and activation of oncogenic pathways such as Wnt/??-catenin and MAPK signaling. Their genetic background and ease of manipulation make them a preferred substrate for studying HCC biology and for validating candidate therapeutic targets.
DNAJA1, a member of the DNAJ/HSP40 family, functions as a co-chaperone for Hsp70 proteins (HSPA1A, HSPA8), facilitating client recognition and ATP-driven folding, trafficking, or degradation. By coupling client binding to Hsp70??s ATPase cycle, DNAJA1 stabilizes the chaperone?Csubstrate complex. Its expression is upregulated by HSF1 in response to cellular stress??heat shock, hypoxia, or TNF-????and it physically interacts with the Hsp70 machinery, STUB1, BAG3, and the IKK complex. DNAJA1 orchestrates the stability of key signaling effectors, including AKT and ??-catenin, thereby positively regulating MAPK, Wnt, and NF-??B pathways. Knockout of DNAJA1 disrupts this chaperone?Cclient network, leading to destabilization of AKT and ??-catenin, diminished downstream signaling, and compromised proliferative and survival capacity.
In the SK-HEP-1 hepatocellular carcinoma background, DNAJA1 loss attenuates protumorigenic signaling nodes, mirroring its described roles in gastric and lung adenocarcinoma. The polyclonal knockout phenotype recapitulates the consequences of impaired Hsp70 co-chaperone activity seen in various cancers, while also providing a tool to explore DNAJA1??s contributions to neurodegenerative processes like Parkinson??s disease, where proteostasis defects are central. Moreover, the model is relevant for studying viral infection, as DNAJA1 has been implicated in viral life cycles.
Investigators can use this polyclonal knockout model for detailed chaperone biology studies involving co-immunoprecipitation and western blotting of phospho-AKT and ??-catenin. Functional assays such as MTT, apoptosis detection, and transwell migration enable phenotypic characterization. The cells are also suitable for RNA-seq-based transcriptomics and Hsp70 inhibitor sensitivity testing, supporting translational research and drug discovery. For additional information, please contact Ascent Research.
