The ECI2 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from AGS gastric epithelial cells, designed to disrupt the ECI2 gene. This polyclonal pool offers a heterogeneously edited cell population, minimizing clonal selection bias while enabling robust functional studies. The targeted disruption of ECI2, encoding mitochondrial enoyl-CoA delta isomerase, allows investigation of fatty acid ??-oxidation in a gastric cancer context.
The parental AGS cell line was established from a 54-year-old female with gastric adenocarcinoma and serves as a well-characterized in vitro model for gastric cancer research. It is widely used to study Helicobacter pylori infection, drug responses, and adenocarcinoma biology, providing a relevant epithelial background for metabolic studies.
ECI2 catalyzes isomerization of 3-cis to 2-trans unsaturated fatty acyl-CoA intermediates, an essential auxiliary step in mitochondrial fatty acid ??-oxidation. This enzyme operates downstream of very long-chain acyl-CoA dehydrogenase (ACADVL) and the mitochondrial trifunctional protein (HADHA/HADHB), and closely interacts with enoyl-CoA hydratase (ECHS1), 3-hydroxyacyl-CoA dehydrogenase (HADH), and 3-ketoacyl-CoA thiolase (ACAA2) to process unsaturated fatty acids. ECI2 transcription is upregulated by PPAR?? and PGC-1?? in response to fatty acid availability and AMPK signaling. Genetic disruption leads to accumulation of partially oxidized intermediates, reduced acetyl-CoA, NADH, and FADH? output, and consequent alterations in TCA cycle activity and redox balance, potentially increasing reactive oxygen species.
In the AGS gastric adenocarcinoma model, metabolic reprogramming towards enhanced lipid utilization supports tumor growth. ECI2 knockout impairs the efficient degradation of dietary and membrane-derived unsaturated fatty acids, creating a metabolic bottleneck that can sensitize cells to nutrient stress and expose targetable vulnerabilities. This model allows detailed dissection of how gastric cancer cells maintain bioenergetic homeostasis when mitochondrial ??-oxidation is compromised, potentially impacting ATP generation, mitochondrial respiration, and cell survival.
Researchers employ this polyclonal knockout population to investigate lipid metabolic dependencies, to screen for inhibitors of fatty acid oxidation, and to study mechanisms of metabolic adaptation in cancer. The cells are well-suited for quantitative assays such as 1?C-oleate-based fatty acid oxidation measurements, Seahorse mitochondrial stress tests, RT-qPCR profiling of FAO gene expression, immunoblotting for mitochondrial proteins, and ATP bioluminescence assays. The polyclonal format provides a robust, cost-effective tool for initial functional genomics studies without clonal bias. For further details, please contact Ascent Research.
