The ECI1 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human HeLa cell line, designed to disrupt the ECI1 gene. This heterogeneous cell pool provides a robust loss-of-function model for investigating mitochondrial fatty acid ??-oxidation of unsaturated fatty acids without the biases introduced by clonal selection.
HeLa cells are a human cervical adenocarcinoma epithelial line, immortalized by HPV18 integration and characterized by aneuploidy and rapid proliferation. They are widely utilized in cancer biology, viral transformation, and cell signaling research due to their well-documented genomic and phenotypic properties. The metabolic adaptability of HeLa cells makes them an ideal host for studying lipid metabolism and mitochondrial dysfunction.
ECI1 encodes enoyl-CoA delta isomerase 1, a mitochondrial enzyme that catalyzes the conversion of 3-cis-enoyl-CoA and 3-trans-enoyl-CoA intermediates to 2-trans-enoyl-CoA, a critical step in the ??-oxidation of unsaturated fatty acids such as oleate and linoleate. This reaction is essential for subsequent processing by ECHS1 (crotonase), the mitochondrial trifunctional protein (HADHA/HADHB), and acyl-CoA dehydrogenases (ACADVL, ACADM, ACADS), ultimately yielding acetyl-CoA, NADH, and FADH2. ECI1 expression is under the transcriptional control of PPARA agonists (fatty acids, fibrates) and the coactivator PGC1A. Gene disruption leads to the accumulation of 3-cis-enoyl-CoA species, impairing acetyl-CoA and ATP production from lipid substrates and forcing a metabolic shift toward glucose and amino acid oxidation, which alters lipid homeostasis and signaling networks.
In the HeLa background, which maintains both glycolytic and oxidative capacities, ECI1 knockout amplifies glycolytic dependency, mimicking metabolic adaptations observed in many cancers. This model enables the study of fatty acid oxidation disorders such as enoyl-CoA delta isomerase deficiency, metabolic cardiomyopathy, and hypoketotic hypoglycemia. Additionally, it provides a platform for exploring how disrupted unsaturated fatty acid metabolism influences cancer cell proliferation, survival, and response to metabolic stress.
Researchers can employ this polyclonal pool in diverse functional assays, including 14C-oleate oxidation to measure fatty acid utilization, LC-MS/MS acylcarnitine profiling to detect metabolic intermediates, and Seahorse metabolic flux analysis to assess mitochondrial oxygen consumption. Complementary techniques include ATP production assays, RT-qPCR and western blotting for ??-oxidation enzymes, and Oil Red O staining for lipid accumulation. This product supports applications from drug screening for lipid metabolism modulators to modeling mitochondrial dysfunction. For technical inquiries and ordering information, please contact Ascent Research.
