The DNAJC10 Knockout SK-HEP-1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human liver adenocarcinoma cell line. This pooled population carries targeted disruption of the DNAJC10 gene (encoding ERdj5), a critical ER co-chaperone. The polyclonal format provides a heterogeneous loss-of-function model, avoiding clonal biases. It is designed for researchers studying ER protein quality control and stress signaling in a hepatocellular carcinoma context.
SK-HEP-1 is an epithelial-derived cell line originally isolated from liver adenocarcinoma ascitic fluid. Widely used as a hepatocellular carcinoma model, it exhibits rapid proliferation and sensitivity to ER stress-inducing agents such as tunicamycin and thapsigargin. These cells endogenously express the core UPR and ERAD machinery, including BiP/GRP78, IRE1??, and components of the SEL1L-HRD1 retrotranslocation complex, providing a relevant platform for dissecting ER proteostasis in transformed hepatocytes.
DNAJC10/ERdj5 is an ER co-chaperone with oxidoreductase activity that reduces disulfide bonds in misfolded glycoproteins, enabling their retrotranslocation and proteasomal degradation via ERAD. ERdj5 is transcriptionally induced by ER stress through IRE1??/XBP1 and ATF6 pathways. It directly interacts with EDEM1, the SEL1L-HRD1 ubiquitin ligase complex, and the p97/VCP ATPase to facilitate substrate extraction. Loss of DNAJC10 disrupts this process, leading to accumulation of misfolded ERAD substrates and sustained UPR signaling, which can shift the balance toward apoptosis through the PERK/ATF4/CHOP axis.
In the SK-HEP-1 liver adenocarcinoma setting, DNAJC10 knockout severely compromises ERAD capacity, resulting in elevated basal ER stress and accumulation of misfolded glycoproteins. This genetic perturbation sensitizes the cells to pharmacological ER stress inducers such as tunicamycin and thapsigargin, while also potentially affecting basal proliferation, migration, and apoptotic thresholds. The polyclonal knockout population faithfully captures the heterogeneity of CRISPR editing outcomes, providing a robust system to explore how ERdj5 deficiency reshapes hepatocellular carcinoma cell behavior and to identify synthetic lethal interactions with other ER stress modulators.
This knockout model is suited for a comprehensive range of experimental approaches including Western blotting for UPR markers (e.g., BiP, CHOP), RT-qPCR for ER stress target genes, and immunofluorescence to visualize ER morphology and protein aggregation. Functional assays such as flow cytometry for apoptosis, cell viability profiling, and transwell migration/invasion can delineate the role of ERdj5 in tumor cell dynamics. Co-immunoprecipitation experiments can probe interactions with EDEM1, HRD1, and p97/VCP. Combined with ER stress induction and drug sensitivity screens, the model facilitates ERAD mechanism studies, hepatocellular carcinoma research, and cancer drug discovery targeting ER proteostasis. For further technical details or to place an order, please contact Ascent Research.
