This CRISPR/Cas9-edited polyclonal knockout cell population targets DNAJC16 in the HGC-27 gastric adenocarcinoma cell line. It comprises a heterogeneous pool of cells with CRISPR/Cas9-mediated disruption of the DNAJC16 gene, generating a loss-of-function model for chaperone biology. As a polyclonal population, these cells reflect the diversity of editing outcomes and are suitable for pooled analyses. They are an ideal system for studying DNAJC16 function in gastric cancer.
The HGC-27 cell line, derived from metastatic gastric adenocarcinoma, is a standard model for human gastric cancer. It retains key features of gastric adenocarcinoma, including dysregulated signaling and stress responses. HGC-27 cells are particularly useful for investigating the unfolded protein response (UPR) and endoplasmic reticulum (ER) stress, which are frequently disrupted in cancers. Their origin makes them a relevant host to examine tumor-specific roles of protein quality control factors like DNAJC16.
DNAJC16 encodes a J-domain co-chaperone that stimulates the ATPase activity of Hsp70 family proteins, including HSPA1A and HSPA5 (BiP). This activity is critical for protein folding, translocation, and degradation. DNAJC16 interacts with Hsp70 members, BAG co-chaperones, HOP (STIP1), and the CHIP E3 ubiquitin ligase. Upstream, its expression is controlled by HSF1 and ER stress sensors IRE1, PERK, and ATF6. By regulating Hsp70, DNAJC16 facilitates client protein triage to folding pathways or proteasomal degradation via CHIP. Disruption of this gene likely impairs protein quality control, leading to proteotoxic stress and UPR activation.
In HGC-27 gastric adenocarcinoma cells, DNAJC16 loss is expected to reduce protein folding capacity, sensitizing cells to proteotoxic stress and altering proliferation and survival. Cancer cells with high stress may depend on Hsp70 co-chaperones for growth, and DNAJC16 knockout can uncover these vulnerabilities. This model allows dissection of how DNAJC16 contributes to UPR and apoptosis regulation. It also provides a platform to study adaptation to chaperone network disruption, with potential therapeutic implications for targeting protein homeostasis in gastric cancer.
Researchers can use this knockout model to assess DNAJC16 in gastric cancer cell proliferation and apoptosis via MTT and caspase activation assays. Co-immunoprecipitation of Hsp70 and Western blotting for Hsp70 ATPase activity monitor chaperone interactions and function. UPR target gene expression can be measured by RT-qPCR, and protein aggregation can be detected by immunofluorescence. The polyclonal population supports pooled functional screens to deconvolute chaperone-mediated processes in cancer. For technical details, please contact Ascent Research.
