The DNAJC3 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population derived from the human liver adenocarcinoma cell line SK-HEP-1, featuring a targeted disruption of the DNAJC3 gene. This polyclonal knockout model serves as a versatile tool for dissecting the roles of the DNAJC3-encoded co-chaperone P58IPK in cellular stress responses, particularly the unfolded protein response (UPR) and translational regulation, within a hepatocyte-derived carcinoma background.
SK-HEP-1 is an established human liver adenocarcinoma cell line that serves as a widely used in vitro model for hepatocellular carcinoma. It exhibits characteristic features of hepatocyte-derived cancer cells, including susceptibility to endoplasmic reticulum stress and dependence on adaptive UPR signaling for survival. This cell line is commonly employed in studies of liver cancer biology, drug resistance, and cellular stress pathways, making it a relevant host for interrogating the functional consequences of DNAJC3 loss.
DNAJC3 encodes P58IPK, an ER-resident co-chaperone that functions as a critical inhibitor of the eIF2?? kinases PERK (EIF2AK3) and PKR (EIF2AK2). Under basal and ER stress conditions, P58IPK interacts with the major ER chaperone BiP (HSPA5) and directly binds to the kinase domains of PERK and PKR, attenuating their activity and limiting phosphorylation of the translation initiation factor eIF2??. DNAJC3 expression is transcriptionally induced by the UPR sensors ATF6 and XBP1, and by interferon-alpha signaling. Knockout of DNAJC3 removes a key brake on eIF2?? phosphorylation, leading to sustained activation of the integrated stress response, elevated ATF4 and CHOP levels, and enhanced apoptosis under conditions of unresolved ER stress.
In SK-HEP-1 liver adenocarcinoma cells, loss of DNAJC3 disrupts the delicate balance of the UPR and renders the cells hypersensitive to ER stress-inducing agents. Given the high secretory burden of hepatocyte-derived cells and the frequent upregulation of UPR components in liver cancer, this polyclonal knockout model enables researchers to dissect how deregulated translational control impacts tumor cell survival, proliferation, and therapeutic response. It provides a platform for evaluating the contribution of the PERK-eIF2??-ATF4-CHOP axis to apoptosis induction in a hepatic cancer setting.
This polyclonal knockout product is suitable for a range of experimental applications, including but not limited to: monitoring UPR activation via Western blot analysis of phospho-eIF2??, ATF4, and CHOP; assessing ER stress sensitivity using tunicamycin or thapsigargin challenge assays; measuring global protein synthesis rates by puromycin incorporation; performing apoptosis assays (e.g., Annexin V staining); and analyzing protein-protein interactions through co-immunoprecipitation of PERK or BiP. The model is relevant to research areas such as ER stress biology, cancer cell signaling, translation regulation, diabetes, and neurodegeneration. For further information, please contact Ascent Research.
