The DNAJC10 Knockout HGC-27 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DNAJC10 gene in the human gastric carcinoma HGC-27 cell line. The polyclonal format provides a heterogeneous pool of edited cells, enabling robust assessment of gene disruption effects without the limitations of single-cell cloning. This knockout model relies on CRISPR/Cas9-mediated gene disruption to create a loss-of-function system for investigating DNAJC10’s roles in endoplasmic reticulum (ER) proteostasis and ER-associated degradation (ERAD).
The HGC-27 host cell line is derived from a poorly differentiated gastric adenocarcinoma and serves as a well-characterized model for gastric epithelial biology and gastric cancer pathogenesis. These cells retain key features of cancerous gastric epithelia, including alterations in mucosal barrier function and secretory capacity, and are widely employed to study tumor cell proliferation, migration, and drug responses. The gastric cancer context is particularly pertinent given the high reliance of secretory tumor cells on ER quality control mechanisms for survival.
DNAJC10 encodes an ER-resident co-chaperone and disulfide reductase that critically maintains ER proteostasis by reducing disulfide bonds in misfolded glycoproteins, thereby facilitating their retrotranslocation and degradation via the HRD1 (SYVN1)-mediated ERAD pathway. DNAJC10 functions in concert with BiP/GRP78 (HSPA5) and interacts with ERAD components including EDEM1, SEL1L, and VCP/p97, ultimately targeting substrates for proteasomal degradation. Upstream, the transcription factors ATF6, XBP1, and ATF4 upregulate DNAJC10 during ER stress to augment ERAD capacity. Knockout of DNAJC10 impairs disulfide reduction and ERAD efficiency, causing accumulation of misfolded proteins and activating the unfolded protein response (UPR) through sensors like IRE1??, which splices XBP1 mRNA.
In the context of HGC-27 gastric carcinoma cells, DNAJC10 knockout is expected to compromise ERAD, leading to constitutive ER stress and altered UPR signaling that may impact tumor cell growth, apoptosis, and chemosensitivity. Given that gastric cancer cells often exhibit heightened ER stress due to secretory demands and oncogenic activation, this model allows dissection of how ER proteostasis pathways contribute to malignancy and therapy resistance. The knockout also provides a tool to examine the interplay between DNAJC10 and other ER chaperones and folding enzymes in maintaining the functional integrity of gastric epithelial cells under physiological and stressful conditions.
This polyclonal knockout cell pool is suited for applications including investigation of ER stress and UPR dynamics by Western blotting and RT-qPCR of UPR markers, immunofluorescence analysis of ER morphology, and sensitivity profiling with ER stress inducers such as tunicamycin. Researchers can perform flow cytometry-based apoptosis assays, MTT cell viability assessments, ubiquitinated protein immunoblotting to track ERAD substrate accumulation, and Transwell migration assays to evaluate metastatic behavior. The model supports gastric cancer biology studies, ER stress-related disease modeling, and drug discovery efforts targeting proteostatic mechanisms. For additional technical details or custom requests, please contact Ascent Research.
