The DNAJC5 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the DNAJC5 gene in human embryonic kidney HEK293T cells. This product provides a pooled, non-clonal loss-of-function model for investigating the cellular roles of DNAJC5, a co-chaperone involved in chaperone-mediated regulation of exocytosis. The polyclonal format ensures representation of diverse editing events across the cell population, making it suitable for bulk population studies where a broad spectrum of knockout efficiencies is acceptable. This knockout model is designed to support mechanistic studies of DNAJC5 function without the confounding effects of neuronal-specific factors.
HEK293T cells are a widely used adherent cell line derived from human embryonic kidney, transformed with adenovirus type 5 and the SV40 large T antigen. This immortalized cell line is renowned for its high transfection efficiency and robust protein expression capacity, making it a preferred host for recombinant protein production, lentivirus packaging, and transient transfection experiments. The epithelial origin of HEK293T cells provides a non-neuronal context in which to study the fundamental roles of DNAJC5, particularly those related to constitutive and regulated secretion pathways that share molecular machinery with neuronal exocytosis.
DNAJC5 (also known as cysteine string protein alpha, CSP??) functions as a co-chaperone that recruits Hsc70 to the SNARE complex, facilitating chaperone-mediated assembly and function of the exocytotic machinery. Its activity is modulated by upstream regulators including palmitoylation by DHHC enzymes, which anchors it to vesicle membranes, and is influenced by calcium signaling. DNAJC5 interacts with key exocytosis factors such as SNAP-25, syntaxin 1A, synaptotagmin, and Hsp40, and its downstream effects include promotion of SNARE complex assembly, regulated exocytosis, and neurotransmitter release. Knockout of DNAJC5 disrupts this chaperone network, impairing proper vesicular trafficking and fusion events. In the HEK293T background, this disruption provides a tractable system to dissect the chaperone-dependent steps of exocytosis, independent of the specialized synaptic environment.
The use of HEK293T cells for DNAJC5 knockout creates a valuable model to study the gene’s role in secretory pathways outside the neuronal context, which is particularly relevant for understanding its involvement in non-neuronal exocytotic processes. DNAJC5 mutations are linked to neuronal ceroid lipofuscinosis type 4 (CLN4, Batten disease), a fatal neurodegenerative disorder. By studying DNAJC5 loss in HEK293T cells, researchers can delineate core chaperone functions that may contribute to the disease pathology, such as failure of vesicle recycling and protein trafficking, without neuronal activity confounders. This model enables systematic investigation of how palmitoylation-dependent membrane localization and co-chaperone interactions are altered upon DNAJC5 disruption.
This polyclonal knockout cell pool is ideal for applications including investigation of chaperone-mediated exocytosis mechanisms, study of DNAJC5-related neurodegeneration, and protein trafficking research. It supports diverse experimental workflows such as western blotting to validate DNAJC5 protein loss, co-immunoprecipitation to assess impaired interactions with SNAP-25 or syntaxin 1A, immunofluorescence to monitor subcellular distribution, and vesicle release assays to quantify exocytosis deficits. Flow cytometry with exocytosis markers and RT-qPCR for SNARE component expression further enable functional screening. For drug discovery efforts targeting Batten disease, these cells offer a scalable platform for compound library screening. For more information, contact Ascent Research.
