The DNAJB2 Knockout SK-HEP-1 Polyclonal Cells product is a heterogeneous population of SK-HEP-1 human hepatocellular carcinoma cells that have undergone CRISPR/Cas9-mediated disruption of the DNAJB2 gene. This polyclonal knockout pool serves as a loss-of-function model for investigating the biological functions of DNAJB2, a J-domain co-chaperone. By avoiding single-cell cloning, the polyclonal format captures a broad spectrum of editing outcomes, minimizing clonal artifacts and enabling robust, reproducible studies in a genetically diverse cell population.
The parental SK-HEP-1 cell line is an epithelial cell line originally derived from the ascitic fluid of a patient with adenocarcinoma of the liver. It has been widely adopted as a model for hepatocellular carcinoma, supporting research into tumor biology, metastatic progression, and xenobiotic metabolism. SK-HEP-1 cells retain key characteristics of hepatic adenocarcinoma, including active signaling pathways relevant to cancer cell stress responses, making them particularly suitable for dissecting the role of chaperone networks in liver cancer.
DNAJB2 encodes a J-domain co-chaperone that stimulates the ATPase activity of HSP70 (HSPA1A), facilitating protein folding and, together with the E3 ubiquitin ligase CHIP (STUB1), targeting misfolded clients to the ubiquitin-proteasome system. Its expression is regulated by HSF1 under heat shock, oxidative stress, and ER stress, with ER stress sensors (IRE1, PERK, ATF6) also contributing. Downstream, DNAJB2 modulates the UPR, affecting targets such as BiP and CHOP, and intersects with ATF4 and XBP1 pathways. DNAJB2 thus coordinates proteostasis with HSP70, BAG co-chaperones, and the proteasome.
In the context of SK-HEP-1 hepatic adenocarcinoma cells, DNAJB2 knockout is expected to disrupt the HSP70 chaperone cycle, resulting in the accumulation of ubiquitinated proteins and heightened sensitivity to proteotoxic insults. Hepatocellular carcinoma cells frequently rely on elevated chaperone activity to cope with intrinsic ER stress, so this model may uncover vulnerabilities exploitable by chaperone-targeted therapies. The knockout enables dissection of how DNAJB2 deficiency alters the balance between pro-survival UPR signaling and stress-induced apoptosis, particularly in scenarios involving chemotherapeutic agents or proteasome inhibition.
This knockout model is suited for Western blotting for HSP70 and ubiquitinated proteins, RT-qPCR for UPR markers such as BiP and CHOP, cell viability assays under ER stress induction, immunofluorescence detection of protein aggregates, and proteasome activity measurements. It also supports migration and invasion studies in a liver cancer background and drug sensitivity testing with proteasome inhibitors like bortezomib. These applications enable research into proteostasis, cancer cell stress adaptation, and chaperone-targeted therapeutic strategies. For additional information, please contact Ascent Research.
