The ECHDC3 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of AGS gastric adenocarcinoma cells with disrupted ECHDC3. This loss-of-function model provides a powerful tool for investigating the role of ECHDC3, a mitochondrial enoyl-CoA hydratase critical for fatty acid beta-oxidation. The polyclonal format preserves genetic heterogeneity, avoiding bias from clonal selection, and supports robust functional studies of lipid metabolism and energy homeostasis in a gastric cancer context.
Derived from human gastric adenocarcinoma, the AGS cell line is a well-characterized epithelial model that maintains gastric mucosal barrier and secretory functions, making it highly relevant for gastric cancer research. These cells exhibit an enhanced reliance on fatty acid oxidation to meet bioenergetic demands, providing an ideal system to dissect metabolic dependencies. The ECHDC3 knockout in this background permits direct examination of how disruption of mitochondrial fatty acid utilization impacts transformed gastric epithelial cells.
ECHDC3 encodes the mitochondrial enoyl-CoA hydratase that catalyzes the second step of the beta-oxidation cycle, converting enoyl-CoA to 3-hydroxyacyl-CoA, and functions in concert with acyl-CoA dehydrogenase, 3-hydroxyacyl-CoA dehydrogenase, and 3-ketoacyl-CoA thiolase. The gene is transcriptionally activated by PPAR?? and its coactivator PGC-1??, downstream of AMPK signaling. Disruption of ECHDC3 reduces production of acetyl-CoA, NADH, FADH2, and ATP, impairs ketone body synthesis, and places metabolic stress on the cell, allowing researchers to interrogate the PPAR??/PGC?1???CECHDC3 axis and its control over bioenergetic flux.
In AGS gastric cancer cells, fatty acid oxidation serves as a critical fuel source to support proliferation and survival, particularly under nutrient-depleted or hypoxic conditions. Ablation of ECHDC3 disrupts this metabolic pathway, forcing the cells to rewire energy production and potentially sensitizing them to metabolic inhibitors. Consequently, this polyclonal knockout model is invaluable for studying lipid metabolic reprogramming in gastric cancer and for evaluating ECHDC3 as a selective therapeutic vulnerability, with the genetic heterogeneity mirroring that of tumors.
Researchers can utilize this polyclonal knockout cell population in diverse functional assays, including Seahorse extracellular flux analysis to measure fatty acid oxidation-dependent oxygen consumption, [^14C]-palmitate tracing to directly quantify beta-oxidation activity, and ATP luminescence assays to assess energy status. Complementary techniques such as western blotting for beta-oxidation enzymes, RT?qPCR profiling of lipid metabolism genes, and mass spectrometry-based lipidomics offer comprehensive metabolic phenotyping. Cell viability assays under metabolic stress, such as glucose deprivation or treatment with fatty acid oxidation inhibitors, can reveal dependencies that may guide therapeutic strategies. For additional product information and technical support, please contact Ascent Research.
