This product is a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DNAJB12 gene in AGS cells. The polyclonal nature of this knockout model preserves the genetic heterogeneity of the edited population, enabling robust interrogation of DNAJB12 function without clonal selection bias. Using CRISPR/Cas9-mediated gene disruption, this loss-of-function model provides a reliable system to study DNAJB12-dependent cellular processes.
AGS cells are a well-characterized human gastric adenocarcinoma epithelial cell line widely used in cancer biology and drug discovery. Derived from a gastric adenocarcinoma, these adherent cells retain key molecular features of gastric epithelium and are permissive to genetic manipulation, making them an ideal host for investigating genes implicated in gastric tumorigenesis and stress response pathways.
DNAJB12 encodes an ER-resident J-domain co-chaperone that plays a central role in ER-associated degradation (ERAD). Mechanistically, DNAJB12 recruits Hsp70 (HSPA) to misfolded proteins within the ER lumen, facilitating their ubiquitination by HRD1 and other ubiquitin ligases. Subsequently, the ATPase p97/VCP extracts these substrates from the ER membrane for proteasomal degradation by the 26S proteasome. DNAJB12 activity is tightly linked to the unfolded protein response (UPR), as its expression is induced by ER stress sensors IRE1, PERK, and ATF6. Thus, DNAJB12 bridges ER stress sensing and the clearance of aberrant proteins, preventing proteotoxicity and maintaining ER homeostasis.
In gastric cancer, dysregulated ERAD and UPR signaling contribute to tumor adaptation and chemoresistance. DNAJB12 knockout in AGS cells provides a physiologically relevant context to dissect how ER chaperone networks support malignant cell survival under proteotoxic stress. This model enables precise analysis of DNAJB12??s role in gastric adenocarcinoma stress resilience, offering insights into potential therapeutic vulnerabilities associated with ERAD dependency.
Researchers can employ this polyclonal knockout cell population in a variety of experimental contexts. Standard applications include monitoring UPR activation (e.g., BiP and CHOP expression via western blotting), assessing ER stress gene expression by RT-qPCR, measuring cell viability under ER stress inducers, and performing co-immunoprecipitation studies to examine DNAJB12?CHsp70 interactions. Additionally, it is suitable for proteasome activity assays and high-content screens to identify modulators of ERAD. For further information or customized services, please contact Ascent Research.
