The ECI1 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell pool with targeted disruption of the ECI1 gene in the Huh-7 human hepatocellular carcinoma cell line. This polyclonal population provides a heterogeneous loss-of-function model, enabling researchers to assess ECI1-dependent phenotypes without clonal selection bias. ECI1 encodes a key mitochondrial enoyl-CoA isomerase necessary for complete ??-oxidation of unsaturated fatty acids, making this model particularly valuable for investigations into lipid metabolism and metabolic reprogramming in liver cancer.
The Huh-7 host cell line was derived from a hepatocellular carcinoma of a 57-year-old Japanese male and is widely utilized as a model for hepatocyte biology, liver metabolism, and HCC. Huh-7 cells retain hepatocyte-like features including expression of metabolic enzymes and susceptibility to hepatotropic viruses, making them suitable for studying hepatic lipid handling, steatosis, and drug metabolism. Their adherent growth and compatibility with various functional assays, such as lipid staining and mitochondrial respiration measurements, further enhance their utility for metabolic research.
ECI1 catalyzes the isomerization of 3-cis and 3-trans double bonds in unsaturated fatty acyl-CoA esters, an essential step permitting their complete degradation via mitochondrial ??-oxidation. ECI1 expression is transcriptionally regulated by lipid-sensing nuclear receptors PPAR?? and PPAR??, as well as by SREBP1c. The enzyme functions within a multienzyme complex that includes very long-chain acyl-CoA dehydrogenase (ACADVL), enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and the mitochondrial trifunctional protein subunits HADHA and HADHB. ECI1 activity generates acetyl-CoA and ATP, thereby supporting mitochondrial respiratory chain function, and interacts with the auxiliary enzyme DECR1 and the peroxisomal EHHADH.
In the Huh-7 hepatocellular carcinoma context, ECI1 disruption impairs unsaturated fatty acid catabolism, likely leading to accumulation of unusual enoyl-CoA intermediates, lipid droplet formation, and lipotoxic stress. This metabolic defect causes a shift in energy homeostasis and may exacerbate non-alcoholic fatty liver disease phenotypes or promote metabolic reprogramming in HCC. The model therefore enables dissection of how defects in mitochondrial unsaturated fatty acid handling contribute to liver cancer progression, steatosis, and metabolic syndrome, offering a physiologically relevant platform for studying lipid-related pathologies.
This polyclonal knockout pool is suited for a range of applications including analysis of unsaturated fatty acid metabolism, investigation of lipid-mediated signaling in cancer cell proliferation, and assessment of steatosis and lipotoxicity. Compatible assays include fatty acid oxidation measurements, Oil Red O staining, ATP luminescence, Seahorse respirometry, metabolomics, lipidomics, and proliferation assays (MTT/BrdU). Western blot and RT-qPCR can be used to monitor changes in fatty acid metabolic genes such as CPT1A, ACC, and PPAR?? targets. For further information, please contact Ascent Research.
