The ECHS1 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human A-549 lung adenocarcinoma line (Homo sapiens), carrying gene disruption at the ECHS1 locus. This heterogeneous pool of edited cells offers a population-level loss-of-function model, reducing clonal selection artefacts for robust functional analysis of mitochondrial fatty acid ??-oxidation and branched-chain amino acid catabolism in vitro.
The A-549 host cell line, isolated from a 58-year-old male with lung carcinoma, is an adherent epithelial line exhibiting alveolar type II characteristics. Widely employed in cancer metabolism, drug metabolism, and viral infection studies, these cells provide a physiologically relevant context for exploring mitochondrial pathways and energy homeostasis dysregulation.
ECHS1 encodes the mitochondrial enzyme enoyl-CoA hydratase, catalyzing the hydration of enoyl-CoA thioesters in the ??-oxidation of fatty acids and degradation of branched-chain amino acids. Its transcription is governed by PPARA, PPARG, PPARGC1A, and HNF4A. ECHS1 interacts with HADHA, HADHB, and ECI1 within the mitochondrial trifunctional protein complex, operating upstream of acetyl-CoA, NADH, and ATP generation. Disruption of ECHS1 impairs these processes, causing diminished mitochondrial respiration and accumulation of metabolic intermediates that trigger mitochondrial dysfunction, resembling ECHS1 deficiency disorders such as Leigh syndrome and metabolic acidosis.
In A-549 adenocarcinoma cells, ECHS1 knockout disrupts oxidative energy metabolism, forcing metabolic rewiring that reveals vulnerabilities under glucose deprivation or nutrient stress. This model is particularly suited for investigating how cancer cells adapt to defective fatty acid oxidation, identifying compensatory metabolic pathways, and evaluating synthetic lethal interactions. It also serves as a platform for studying mitochondrial dysfunction linked to encephalopathies.
These polyclonal cells are compatible with a variety of functional assays, including Seahorse mitochondrial stress testing, palmitate oxidation measurement, western blotting, RT-qPCR, and metabolomic profiling. Research applications include metabolic reprogramming analysis, fatty acid oxidation inhibitor screening, Leigh syndrome disease modeling, and validation of ECHS1-related mitochondrial dysfunction. For further details, contact Ascent Research.
