The ECHDC1 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population based on the Huh-7 hepatocellular carcinoma line. This loss-of-function model, generated by CRISPR/Cas9-mediated gene disruption of ECHDC1, provides a versatile tool for investigating mitochondrial fatty acid metabolism in a liver cancer context.
The Huh-7 host line is a well-differentiated hepatocyte-derived carcinoma originally from a 57-year-old Japanese male liver tumor. It retains hepatocytic functions including active lipid metabolism, making it an appropriate system for studying fatty acid ??-oxidation and its dysregulation in cancer.
ECHDC1 encodes a mitochondrial enoyl-CoA hydratase that catalyzes the hydration of trans-2-enoyl-CoA to L-3-hydroxyacyl-CoA within the fatty acid ??-oxidation spiral. Its expression is regulated by PPAR?? and PGC-1??, downstream of insulin/glucagon signaling. ECHDC1 functions in association with HADHA, HADHB, and electron transfer flavoprotein (ETF), linking its activity to the TCA cycle through generation of acetyl-CoA, NADH, and FADH2. Upstream components include CPT1, CPT2, and acyl-CoA dehydrogenases, while downstream steps involve HADH and ACAA2. Knockout of ECHDC1 disrupts fatty acid degradation, causing accumulation of medium-chain fatty acids and intermediates, and altering cellular energy and lipid homeostasis.
In Huh-7 cells, ECHDC1 knockout impairs mitochondrial ??-oxidation, revealing dependencies on fatty acid utilization typical of hepatocellular carcinoma. This model facilitates studies on metabolic reprogramming, lipotoxicity, and the intersection of fatty acid oxidation defects with liver cancer biology. It also serves as a cellular platform for investigating ethylmalonic encephalopathy-related pathways.
Researchers can use these cells for fatty acid oxidation assays with radiolabeled palmitate, metabolomic profiling of acylcarnitines, and Seahorse-based mitochondrial respiration measurements. The model supports screening for lipid metabolism modulators, analysis of lipid accumulation via Oil Red O staining, and cell viability tests under nutrient stress. Gene disruption can be verified by western blotting and RT-qPCR. For further information, contact Ascent Research.
