The ECI2 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population characterized by targeted disruption of the ECI2 gene in HeLa cervical adenocarcinoma epithelial cells. This heterogeneous pool of loss-of-function mutants provides a genetically diverse model system for investigating ECI2 deficiency, avoiding clonal selection artifacts and enabling robust functional genomics approaches.
HeLa cells, an immortalized human cell line originating from cervical adenocarcinoma, are extensively utilized in biomedical research, particularly in cancer biology and metabolism, due to their rapid proliferation and well-documented signaling networks. Their adaptation to culture conditions and genetic malleability make them a reliable host for dissecting mitochondrial lipid metabolism pathways.
ECI2 encodes enoyl-CoA delta isomerase 2, a mitochondrial auxiliary enzyme that catalyzes the isomerization of 3-cis and 2-trans unsaturated fatty acyl-CoA intermediates to the 2-trans form, a prerequisite for their complete beta-oxidation. Within the mitochondrial fatty acid oxidation machinery, ECI2 physically and functionally interacts with HADHA, HADHB, ACADVL, and ECHS1. Its expression is transcriptionally regulated by PPARA, PPARD, and PPARGC1A, and its activity is modulated by AMPK and SIRT1 in response to cellular energy status. By enabling the degradation of unsaturated fatty acids, ECI2 drives acetyl-CoA generation, ATP synthesis, and mitochondrial respiration, while suppressing lipid droplet accumulation.
In the context of HeLa cells, knockout of ECI2 is expected to impair the complete oxidation of unsaturated fatty acids, resulting in the buildup of isomerized acyl-CoA intermediates and diminished lipid-derived ATP production, especially when glucose availability is limited. This metabolic defect likely perturbs mitochondrial membrane potential, elevates lipid droplet accumulation, and reduces proliferative capacity under conditions that require fatty acid utilization. Consequently, this model serves as a valuable tool for examining how mitochondrial beta-oxidation supports cancer cell metabolic flexibility and contributes to oncogenic metabolism.
Researchers can employ these polyclonal knockout cells to perform fatty acid oxidation assays using radiolabeled oleate, Seahorse mitochondrial stress tests, and lipid droplet visualization with BODIPY or Nile Red staining. Additional applications include acylcarnitine profiling by mass spectrometry, ATP quantification, proliferation assays under glucose-depleted conditions, and RNA-seq to identify transcriptional adaptations. This product is particularly suited for studying lipid metabolism reprogramming in cervical cancer and screening for metabolic inhibitors targeting fatty acid oxidation. For detailed information or technical support, please contact Ascent Research.
