DNAJC1 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, featuring targeted disruption of the DNAJC1 gene to abolish its expression across a heterogeneous cell pool. This loss-of-function model enables the study of ER proteostasis and stress signaling without clonal selection artifacts, providing a reliable platform for investigating co-chaperone-dependent protein quality control in a cancer-relevant epithelial background.
The parental HeLa cell line originates from human cervical adenocarcinoma and harbors integrated HPV18 sequences, conferring immortalized, epithelial growth characteristics. Its robust proliferation and well-documented sensitivity to ER stress inducers like tunicamycin and thapsigargin make it ideal for examining secretory pathway dysfunction. HeLa cells express core UPR and ERAD components, offering a suitable host for functional interrogation of ER-resident co-chaperones in both basal and stressed conditions.
DNAJC1 is an ER-resident J-domain co-chaperone that recruits and activates BiP (HSPA5) to facilitate protein translocation, folding, and UPR regulation. Its expression is controlled by ER stress sensors ATF6, IRE1/XBP1, and PERK/ATF4, and it directly influences downstream effectors including CHOP (DDIT3) and ERAD ubiquitin ligases. DNAJC1 interacts with the Sec61 translocon, calnexin, and HSP90, positioning it centrally in ER proteostasis. Knockout eliminates BiP co-chaperone activity, impairing folding capacity and severely sensitizing cells to ER stress-induced apoptosis upon exposure to agents that increase misfolded protein loads.
In the HeLa background, DNAJC1 deficiency creates a state of enhanced susceptibility to ER stress, providing a sensitive model for dissecting the IRE1/XBP1, PERK/ATF4, and ATF6 signaling branches. It allows for the examination of how co-chaperone dysfunction influences cell viability under proteotoxic conditions and may uncover links between ER stress adaptation and HPV-mediated oncogenesis, contributing to our understanding of cancer cell resilience against protein misfolding stress.
These polyclonal knockout cells are suitable for a range of assays including Western blot analysis of UPR markers BiP and CHOP, RT-qPCR quantification of XBP1 mRNA splicing, immunofluorescence staining to visualize ER structural changes, and cell viability assays under titrated ER stress conditions. Co-immunoprecipitation studies can confirm disrupted DNAJC1-BiP interaction. The model supports drug screening for proteostasis modulators, apoptosis signaling research, and protein misfolding disease modeling, particularly in the context of cancer cell biology. For further technical inquiries, please contact Ascent Research.
