The DNAJC10 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population in which the DNAJC10 gene has been functionally disrupted. This product provides a heterogeneous pool of HAP1 cells carrying a variety of loss-of-function mutations at the DNAJC10 locus, enabling robust gene knockout studies without the need for clonal selection. The polyclonal format is particularly suited for applications where pooled knockout cells provide a more representative model of gene disruption, avoiding clonal artifacts and simplifying experimental workflows.
The HAP1 cell line is a near-haploid human fibroblast-like cell line derived from the KBM-7 chronic myeloid leukemia line. It exhibits an adherent growth pattern and retains a predominantly haploid karyotype, making it a powerful tool for genetic screening and functional genomics. The haploid nature of HAP1 cells allows for efficient gene targeting and clear phenotypic readouts, as a single allele disruption is sufficient to manifest gene function alterations, which is especially advantageous in knockout studies.
DNAJC10 encodes ERdj5, an ER-resident co-chaperone with disulfide reductase activity. It facilitates ERAD by reducing disulfide bonds in misfolded glycoproteins, promoting their retrotranslocation for proteasomal degradation. DNAJC10 interacts with BiP (GRP78) and EDEM1, and associates with the SEL1L-HRD1 (SYVN1) complex. Activated by XBP1 and ATF6 during ER stress, DNAJC10 targets misfolded proteins such as mutant proinsulin and alpha-1 antitrypsin Z, and attenuates PERK and IRE1 signaling to reduce apoptosis. Thus, it maintains ER proteostasis and calcium homeostasis, and mitigates the unfolded protein response.
In the HAP1 model, DNAJC10 knockout overcomes the limitation of diploid compensation, allowing a clear dissection of ER stress pathways. The near-haploid background ensures that the disruption of DNAJC10 directly affects ER proteostasis, predisposing cells to ER stress sensitivity. This model is invaluable for investigating mechanisms of ER stress-related diseases, including hepatocellular carcinoma, breast cancer, neurodegenerative disorders, and diabetes. It serves as a platform to explore how tumor cells survive under chronic ER stress and to validate DNAJC10 as a potential therapeutic target. Additionally, it enables the study of ERAD components and redox homeostasis in a simplified genetic context.
This knockout population supports diverse assays: western blotting for DNAJC10, BiP, CHOP, cleaved caspase-3; RT-qPCR for XBP1 splicing and ERAD component expression; ER stress induction with tunicamycin or thapsigargin followed by cell viability; proteasome inhibition with MG132 to accumulate ERAD substrates; co-immunoprecipitation of DNAJC10-BiP; and immunofluorescence for ER morphology. These tools enable functional genomics screens, drug target validation, and mechanistic studies of ER stress and the UPR. For more information, contact Ascent Research.
