The EHHADH Knockout AGS Polyclonal Cells product consists of a heterogeneous population of AGS cells that have been subjected to CRISPR/Cas9-mediated gene disruption targeting the EHHADH locus. This polyclonal knockout format provides a genetically diverse pool of cells bearing loss-of-function mutations in the EHHADH gene, enabling robust functional studies without the bottleneck of single-cell clonal selection. The editing process introduces targeted disruptions that ablate EHHADH protein expression, creating a powerful model for investigating the consequences of EHHADH deficiency in a gastric epithelial context.
The AGS cell line is a well-established human gastric adenocarcinoma model derived from the primary tumor tissue of a 54-year-old female patient. AGS cells form adherent monolayers and retain key epithelial characteristics, making them a widely used platform for research on gastric cancer biology, Helicobacter pylori pathogenesis, and gastric epithelial signaling pathways. Their robust growth in culture and susceptibility to genetic manipulation facilitate the generation of knockout derivatives for mechanistic studies.
EHHADH encodes the peroxisomal L-bifunctional enzyme, which is central to the beta-oxidation of long-chain fatty acids. This enzyme catalyzes the sequential hydration of enoyl-CoA to 3-hydroxyacyl-CoA and dehydrogenation to 3-ketoacyl-CoA, yielding acetyl-CoA and NADH. Its expression is transcriptionally regulated by PPARA and PPARD, which are activated by free fatty acids and during fasting, with coactivation by PGC1A. For peroxisomal import, EHHADH interacts with the peroxin proteins PEX5 and PEX14. Within the peroxisome, it functions in a metabolic assembly alongside HSD17B4, ACOX1, SCP2, and ACAA1 to complete the fatty acid degradation cycle.
In AGS gastric epithelial cells, EHHADH likely serves as a critical node linking peroxisomal lipid catabolism to cellular energy status and oxidative stress management. Gastric cancer cells frequently reprogram metabolic pathways to sustain proliferation and survival, and altered fatty acid oxidation is increasingly recognized in tumor metabolism. Disruption of EHHADH in this model provides a means to dissect how peroxisomal beta-oxidation contributes to gastric cancer cell bioenergetics, redox balance, and the response to lipid-rich microenvironments.
This knockout cell population is suited for a variety of experimental applications, including investigations of peroxisomal fatty acid oxidation using metabolic flux analysis, assessment of lipid utilization via fatty acid oxidation assays, and examination of peroxisomal morphology through immunofluorescence staining for peroxisomal markers. It can be employed in drug screening campaigns for metabolic disorders and for exploring the intersection of lipid metabolism with gastric cancer progression. Combined with molecular techniques such as Western blotting and RT-qPCR, researchers can profile the expression of enzymes in the PPAR signaling and bile acid biosynthesis pathways. For further details or custom inquiries, please contact Ascent Research.
