ECHDC3 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the Huh-7 human hepatocellular carcinoma cell line. This heterogeneous pool carries targeted disruption of the ECHDC3 gene, offering a physiologically relevant model for investigating mitochondrial fatty acid metabolism in a liver cancer context. The polyclonal format preserves population-level diversity while abolishing ECHDC3 function, enabling robust functional analyses without clonal selection bias.
The Huh-7 cell line originates from a well-differentiated hepatocyte-derived carcinoma and retains key hepatic traits, including active lipid metabolism and susceptibility to hepatitis C virus infection. It is widely utilized as a hepatocyte surrogate for studying liver-specific metabolic pathways and viral pathogenesis. The hepatocellular background makes Huh-7 particularly suitable for exploring connections between lipid homeostasis and carcinogenesis, as well as for evaluating metabolic-targeted therapeutics.
ECHDC3 encodes a mitochondrial enoyl-CoA hydratase that catalyzes the hydration of trans-2-enoyl-CoA to L-3-hydroxyacyl-CoA in the fatty acid beta-oxidation spiral. This reaction is essential for complete fatty acid degradation, yielding acetyl-CoA for the TCA cycle and reduced cofactors NADH and FADH2 for oxidative phosphorylation. ECHDC3 expression is transcriptionally controlled by PPAR??, SREBP-1, and PGC-1??, which integrate lipid catabolic programs in response to fatty acid levels. The enzyme functions within a multienzyme complex including the mitochondrial trifunctional protein subunits HADHA and HADHB and cooperates with ECHS1, the short-chain enoyl-CoA hydratase. Its activity depends on the carnitine shuttle components CPT1 and CPT2 for substrate import and generates downstream metabolites such as shortened acyl-CoAs, acetyl-CoA, NADH, and FADH2. This central role positions ECHDC3 as a key node in mitochondrial energy production from lipid substrates.
In the Huh-7 background, disruption of ECHDC3 impairs fatty acid beta-oxidation, likely leading to accumulation of unprocessed lipid intermediates and altered energy metabolism. These hepatocellular carcinoma cells depend heavily on lipid catabolism for proliferation, making them an ideal model to examine how fatty acid oxidation defects contribute to metabolic reprogramming in liver cancer. The knockout may exacerbate lipid droplet formation and perturb mitochondrial respiration, offering insights into non-alcoholic fatty liver disease and metabolic syndrome. This model links a specific enzymatic lesion to hepatocellular dysfunction, facilitating genotype-phenotype studies in lipid metabolism disorders.
This polyclonal knockout cell product supports diverse experimental workflows, including Seahorse-based OCR measurements of fatty acid oxidation rates, Oil Red O staining of lipid accumulation, and ATP quantification to assess metabolic output. Researchers can confirm gene disruption via western blotting and RT-qPCR, while metabolomics profiling and cell viability assays reveal global metabolic adaptations. These cells serve as a platform for drug screening in metabolic disorders and for dissecting how lipid metabolism intersects with oncogenic signaling in hepatocellular carcinoma. For detailed technical specifications and ordering information, please contact Ascent Research.
