The DNAJC10 Knockout HeLa Polyclonal Cells product comprises a heterogeneous population of HeLa cells engineered by CRISPR/Cas9-mediated disruption of the DNAJC10 gene locus. This polyclonal knockout cell pool enables loss-of-function studies without clonal isolation, preserving the genetic diversity inherent to the editing process and avoiding biases associated with single-cell clones. The polyclonal format is particularly suited for investigating ER-associated degradation (ERAD) pathway dynamics, as it better reflects the stochastic nature of gene disruption and can reveal population-level responses to ER stress. This product provides a versatile tool for researchers studying protein quality control, unfolded protein response (UPR) signaling, and the role of ERdj5 in cancer and neurodegenerative disease models.
The host HeLa cell line is an immortalized human cervical adenocarcinoma epithelial line originally derived from Henrietta Lacks. HeLa cells are widely employed in biomedical research due to their robust growth, ease of transfection, and well-characterized signaling networks. Their epithelial origin and tumorigenic background make them particularly relevant for investigating ER stress responses in cancer biology, as malignant cells often rely on adaptive UPR mechanisms to survive proteotoxic insults. The HeLa model offers a consistent and reproducible platform for functional genomics studies, including CRISPR-based gene targeting.
DNAJC10 encodes the ER-resident co-chaperone ERdj5, a bifunctional protein possessing both oxidoreductase and co-chaperone activities. ERdj5 directly interacts with misfolded glycoproteins, BiP/GRP78, EDEM1, and OS9, and cooperates with the SEL1L-HRD1 ubiquitin ligase complex to mediate ERAD. Its reductase activity cleaves disulfide bonds in substrates, facilitating retrotranslocation and subsequent ubiquitination by HRD1, followed by proteasomal degradation. This mechanism is tightly regulated by upstream ER stress sensors, including ATF6 and XBP1s, which transcriptionally activate DNAJC10 expression upon accumulation of misfolded proteins. Additionally, ERdj5 activity is influenced by pharmacological ER stress inducers such as tunicamycin and thapsigargin.
In HeLa cells, disruption of DNAJC10 impairs the clearance of misfolded ER proteins, leading to constitutive UPR activation and heightened sensitivity to ER stress. This model is valuable for deciphering how cancer cells modulate ERAD to sustain survival under challenging conditions, including hypoxia, nutrient deprivation, and chemotherapeutic stress. Because HeLa cells exhibit constitutive UPR pathway activity, the knockout phenotype may reveal synthetic lethal interactions with other ERAD components or proteasome inhibitors, offering insights into therapeutic vulnerabilities.
This polyclonal knockout population is ideal for mechanistic studies of ERAD substrate recognition and retrotranslocation, screening for modulators of ERdj5-dependent degradation, and profiling drug sensitivity in the context of ER stress. Representative applications include monitoring UPR markers (BiP, CHOP) by western blotting, quantifying XBP1 splicing via RT-qPCR, assessing viability under tunicamycin with flow cytometry, performing ERAD reporter assays, and evaluating protein half-life through cycloheximide chase experiments. Co-immunoprecipitation can be used to interrogate ERdj5 interactions with HRD1 and EDEM1. For further information or technical support, please contact Ascent Research.
