The IVD Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the SK-HEP-1 human hepatocellular carcinoma cell line. This loss-of-function model targets isovaleryl-CoA dehydrogenase (IVD), a mitochondrial enzyme critical for leucine catabolism. CRISPR/Cas9-mediated gene disruption inactivates IVD across a heterogeneous cell pool, preserving the genetic diversity of the parental line while ensuring absent enzyme activity. The polyclonal format is suitable for studies requiring a representative tumor population with abrogated branched-chain amino acid metabolism.
The SK-HEP-1 parental line, established from ascites of a liver adenocarcinoma patient, exhibits both epithelial and endothelial features and is a standard model for liver cancer research and drug testing. It retains hepatic metabolic properties, making it an ideal host to study IVD loss in a tumor context. Its robust growth and well-characterized signaling support detailed metabolic and oncogenic pathway analysis.
IVD encodes isovaleryl-CoA dehydrogenase, which converts isovaleryl-CoA to 3-methylcrotonyl-CoA in the mitochondrial matrix, a key step in leucine degradation. Regulated by PPARA, HNF4A, and SIRT3, IVD interacts with electron transfer flavoprotein (ETF) and ETF dehydrogenase to feed electrons to the respiratory chain. Downstream, 3-methylcrotonyl-CoA is processed by MCCC1/MCCC2, ultimately generating acetoacetate and acetyl-CoA. IVD disruption causes accumulation of isovaleryl-CoA and isovaleric acid, impairing mitochondrial energy production and organic acid homeostasis.
In SK-HEP-1 cells, IVD knockout models isovaleric acidemia and reveals metabolic vulnerabilities in hepatocellular carcinoma. Loss of IVD forces metabolic reprogramming, potentially altering mitochondrial respiration and fatty acid oxidation, and creates leucine dependency that can be targeted therapeutically. This model is valuable for exploring how liver cancer cells adapt to defective branched-chain amino acid degradation and for testing metabolic rescue strategies.
Applications include disease modeling, metabolic reprogramming studies in liver cancer, and drug screening for mitochondrial dysfunction. Representative assays are LC-MS-based metabolomics for isovalerylglycine and isovalerylcarnitine, fluorometric IVD activity measurement, Seahorse mitochondrial respiration analysis, and Western blotting for IVD protein. Cell viability under leucine deprivation and apoptosis assays further validate the model. For technical inquiries, please contact Ascent Research.
