DNAJC5 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma cell line. This pool contains heterogeneous disruptions of the DNAJC5 gene, enabling loss-of-function studies of the encoded cysteine string protein (CSP) co-chaperone. The cells are generated through CRISPR/Cas9-mediated gene disruption, providing a mixed population suitable for pooled functional screens without clonal isolation. They are maintained under standard adherent culture conditions and serve as a robust model for investigating DNAJC5-related biology.
The AGS host cell line is an epithelial, adherent model of human gastric adenocarcinoma, originally established from a 54-year-old female patient. It is widely utilized in gastric cancer research to study tumor cell proliferation, migration, invasion, and signaling pathways. AGS cells offer a scalable platform for genetic manipulation and downstream phenotypic assays, making them ideal for dissecting gene function in the context of gastric carcinogenesis.
DNAJC5 encodes cysteine string protein, a co-chaperone that stimulates Hsc70 ATPase activity to promote protein folding, complex disassembly, and SNARE-mediated vesicle fusion. It is regulated by phosphorylation via cAMP-dependent protein kinase (PKA) and transcriptionally controlled by heat shock factor 1 (HSF1) under unfolded protein stress. DNAJC5 interacts with Hsc70, SNAP-25, Syntaxin-1, and Synaptotagmin to facilitate SNARE complex assembly, which involves syntaxin, SNAP-25, and VAMP. Downstream, Hsc70 activation by DNAJC5 contributes to exosome biogenesis and protein aggregate clearance. These functions connect DNAJC5 to chaperone-mediated protein folding, SNARE-mediated vesicle fusion, and autophagy-lysosomal pathways, with representative components including Hsc70, DNAJC5, SNARE proteins, HSP40, and HSP90.
In AGS gastric cancer cells, DNAJC5 is implicated in regulating exosome secretion and maintaining proteostasis. Exosomes from cancer cells carry bioactive cargo that promotes tumor progression and microenvironment remodeling. Disruption of DNAJC5 may alter exosome composition and secretion dynamics, providing a model to dissect exosome-mediated tumorigenic mechanisms. Furthermore, its co-chaperone activity is crucial for managing protein misfolding and aggregation, processes often dysregulated in cancer cells and relevant to stress responses and survival.
This polyclonal knockout cell population enables investigation of DNAJC5 roles in gastric cancer, exosome secretion mechanisms, and protein quality control in non-neuronal cells. It is applicable to drug screening for chaperone modulators and modeling protein aggregation dynamics. Experimental approaches include Western blotting, RT-qPCR, exosome isolation with nanoparticle tracking analysis, co-immunoprecipitation, immunofluorescence microscopy, and cell migration, invasion, and viability assays. For additional technical details and ordering information, please contact Ascent Research.
