ECHDC1 Knockout A-549 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout population of A-549 human lung adenocarcinoma cells, designed for loss-of-function studies of the ECHDC1 gene. Unlike clonal cell lines, this heterogeneous population contains diverse editing outcomes, making it ideal for robust functional analyses of mitochondrial fatty acid beta-oxidation without the selection bias of single-cell clones.
The A-549 parental cell line was originally isolated from the lung adenocarcinoma tissue of a 58-year-old male and displays adherent epithelial morphology. As a well-characterized model of non-small cell lung cancer (NSCLC), A-549 cells are particularly suited for cancer metabolism research, given their ability to utilize fatty acids as an energy source under metabolic stress, reflecting the metabolic plasticity of tumor cells.
ECHDC1 encodes a mitochondrial enoyl-CoA hydratase that catalyzes the hydration of trans-2-enoyl-CoA to 3-hydroxyacyl-CoA in the second step of fatty acid beta-oxidation. This reaction is crucial for acetyl-CoA generation and ATP production. ECHDC1 is regulated by PPARA (PPAR??) and PPARGC1A, which are activated by AMPK signaling in response to fatty acid availability. It functions within the beta-oxidation pathway downstream of CPT1A and CPT2, in concert with ACADVL, HADHA, and HADHB of the trifunctional protein complex. Disruption leads to impaired beta-oxidation flux, altered acylcarnitine profiles, and reduced TCA cycle substrates.
In A-549 cells, metabolic reprogramming often involves enhanced lipid metabolism to sustain proliferation. ECHDC1 knockout in this context allows investigation of cancer cell dependency on fatty acid oxidation, especially under glucose-limited conditions. This model helps elucidate how loss of mitochondrial beta-oxidation affects bioenergetics, redox balance, and macromolecular synthesis in lung adenocarcinoma, and is relevant to studies of PPAR signaling and cancer metabolism.
This product supports a variety of experimental approaches. Western blotting and RT-qPCR enable confirmation of ECHDC1 disruption and downstream expression changes. Seahorse XF flux analysis permits direct measurement of mitochondrial oxygen consumption and fatty acid oxidation rates. Acylcarnitine profiling by LC-MS can detect alterations in beta-oxidation intermediates, while proliferation assays under glucose-depleted conditions reveal metabolic vulnerabilities. These polyclonal knockout cells are a valuable tool for functional genomics of lipid metabolism, cancer metabolism, and metabolic disease modeling. For further technical information, please contact Ascent Research.
