The ECI2 Knockout A-549 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population with disruption of the ECI2 gene, which encodes mitochondrial enoyl-CoA delta isomerase 2. This heterogeneous loss-of-function model, derived from the human A-549 lung adenocarcinoma cell line, enables investigation of unsaturated fatty acid oxidation and its metabolic consequences without the bias of clonal selection.
The A-549 parental line originates from human lung adenocarcinoma, displays adherent epithelial morphology, and retains characteristics of alveolar Type II-like cells, including pulmonary surfactant production. Widely used in cancer biology and drug discovery, these cells offer a relevant platform for studying lipid metabolism in non-small-cell lung cancer.
ECI2 catalyzes the isomerization of 3-enoyl-CoA intermediates to 2-trans-enoyl-CoA, a crucial step for complete ??-oxidation of unsaturated fatty acids within mitochondria. It interacts with HADHA, HADHB, ECH1, and electron transfer flavoprotein (ETF) along the fatty acid degradation pathway. Transcriptional control is mediated by PPAR?? and PPAR??, with coactivation by PGC-1?? and modulation by AMPK signaling. ECI2 activity drives production of acetyl-CoA, fueling the TCA cycle and oxidative phosphorylation for ATP synthesis. Disruption of ECI2 thus impairs energy generation from unsaturated lipids and perturbs downstream fatty acid-derived metabolites.
In A-549 cells, ECI2 knockout is expected to hinder degradation of unsaturated fatty acids, which are major substrates in lipid-rich microenvironments. This metabolic bottleneck may reduce bioenergetic capacity, alter lipid homeostasis, and affect proliferation and surfactant synthesis. The model allows examination of how disrupted fatty acid oxidation interacts with PPAR signaling and tumor metabolism, highlighting potential vulnerabilities in lung adenocarcinoma.
Applications include metabolic flux analysis with Seahorse XF Fatty Acid Oxidation Assays, oleate/palmitate oxidation measurements, and lipidomic profiling to map altered lipid species. Researchers can corroborate findings by immunoblotting for ECI2, HADHA, HADHB, and ECH1, performing RT-qPCR for PPAR target genes, and conducting proliferation/survival assays under lipid-rich conditions. These polyclonal knockout cells are suited for validating metabolic drug targets, characterizing fatty acid oxidation defects, and dissecting lipid metabolism in cancer. For further inquiries, please contact Ascent Research.
