DNAJC16 Knockout HEK293T Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T host line, enabling loss-of-function studies of the ER-resident DnaJ (Hsp40) co-chaperone DNAJC16. This product consists of a heterogeneous pool of edited cells with targeted disruption of the DNAJC16 gene, generated by CRISPR/Cas9-mediated gene disruption, and is supplied as a ready-to-use polyclonal population for functional genomics and cell-based assays.
The host HEK293T cell line is a transformed human embryonic kidney epithelial line stably expressing the SV40 large T antigen and containing integrated adenovirus 5 DNA. This genetic background supports high-level transient and stable protein expression and permits episomal replication of SV40 origin-containing plasmids, making the cells a versatile platform for studying protein folding, signaling pathways, and viral production. The epithelial origin and robust growth characteristics further facilitate scalable experimental workflows.
DNAJC16 encodes an ER luminal co-chaperone that directly accelerates the ATPase cycle of BiP/GRP78, the master regulator of ER protein folding. By enhancing BiP??s substrate binding and release cycles, DNAJC16 promotes efficient folding of nascent polypeptides and quality control of misfolded proteins. Its function is integrated into the unfolded protein response (UPR) network: expression is upregulated by ER stress stimuli such as tunicamycin and thapsigargin via the transcription factors ATF4 and XBP1s. Downstream, DNAJC16 acts upstream of IRE1 oligomerization and phosphorylation, XBP1 mRNA splicing, and PERK-eIF2??-ATF4 signaling, and interacts physically with BiP/GRP78, IRE1, and other ER DnaJ proteins to modulate substrate handling. Knockout of DNAJC16 impairs BiP chaperone cycles, leading to chronic ER proteostatic imbalance, constitutive UPR activation, and heightened susceptibility to ER stress-induced apoptosis.
In the HEK293T background, disruption of DNAJC16 creates a powerful model for dissecting the molecular interplay between ER co-chaperones and the three UPR branches. Because HEK293T cells retain robust ER and secretory pathway capacity, the DNAJC16 knockout reveals specific defects in BiP-dependent substrate engagement and downstream signaling thresholds. This model is particularly relevant for research into neurodegenerative diseases, where protein misfolding overwhelms ER quality control; cancer, where tumor cells adapt to ER stress; and metabolic disorders linked to ER dysfunction. Researchers can use these cells to examine how loss of a single co-chaperone redirects UPR outputs and impacts cell fate decisions under pharmacologically induced ER stress.
Key applications include detailed investigation of ER stress and UPR mechanisms, functional characterization of co-chaperone networks, and studies of protein misfolding pathologies. Representative assays using this knockout model include Western blot analysis of UPR markers such as BiP, CHOP, and phospho-eIF2??; RT-qPCR quantification of XBP1 splicing and UPR target genes; immunofluorescence detection of ER stress foci; ERSE-luciferase reporter assays to monitor UPR transcriptional activity; cell viability assays under tunicamycin or thapsigargin challenge; co-immunoprecipitation of BiP complexes; and global transcriptome profiling by RNA-seq. For further technical specifications and customization options, please contact Ascent Research.
