The DNAJC5 Knockout Huh-7 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal cell population derived from the Huh-7 human hepatocellular carcinoma line, engineered for targeted disruption of the DNAJC5 gene. This mixed population provides a versatile loss-of-function model, enabling functional studies without the selection bias of single-cell clones. Disruption of DNAJC5 in Huh-7 cells creates a platform to investigate gene function in a hepatic epithelial context.
The parental Huh-7 line, originating from a liver tumor of a 57-year-old Japanese male in 1982, is a well-differentiated epithelial hepatocellular carcinoma model. Extensively characterized, Huh-7 cells are widely used for liver cancer research, hepatitis C virus studies, and hepatocyte function assays, including lipoprotein metabolism and protein secretion. Their robust growth and genetic tractability make them an ideal host for CRISPR-based knockout generation.
DNAJC5 encodes cysteine string protein alpha (CSP??), a co-chaperone that anchors to synaptic vesicles via palmitoylation and recruits Hsc70 (HSPA8) to drive clathrin uncoating during endocytosis and exocytosis. CSP?? directly interacts with SNARE proteins SNAP-25 and syntaxin-1, preventing their aggregation and preserving synaptic vesicle release. Upstream, palmitoyl acyltransferases and synaptic activity regulate its localization, while downstream it critically maintains clathrin dynamics and vesicle recycling. Mutations in DNAJC5 cause adult-onset neuronal ceroid lipofuscinosis type 4, underscoring its role in protein quality control.
In the Huh-7 hepatocellular carcinoma background, loss of CSP?? likely disrupts clathrin-mediated endocytosis and secretory processes, as Hsc70-dependent chaperone activity is essential for hepatic protein trafficking. Huh-7 cells are highly secretory, producing albumin and lipoproteins, rendering this knockout model particularly relevant for studying chaperone impairments in secretion. This polyclonal population thus bridges neuronal chaperone biology with hepatocyte vesicle dynamics, offering a unique tool to investigate convergent mechanisms of protein aggregation and trafficking defects.
This product is suited for diverse applications including neuronal ceroid lipofuscinosis type 4 research, synaptic vesicle trafficking studies, and chaperone function analysis. Typical assays include western blotting for CSP?? and synaptic proteins, RT-qPCR for transcriptional changes, immunofluorescence for vesicle markers, clathrin-mediated endocytosis assays, and albumin ELISA to monitor secretion. The polyclonal format facilitates rapid phenotypic screening and complementation rescue experiments. For additional information or custom inquiries, please contact Ascent Research.
