The IVD Knockout Jurkat Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population for the human IVD gene in the Jurkat T-lymphocyte background. This product is designed for loss-of-function studies of isovaleryl-CoA dehydrogenase (IVD), a mitochondrial enzyme critical for leucine catabolism. Through CRISPR/Cas9-mediated disruption of the IVD locus, these cells serve as a versatile model to investigate metabolic and cellular consequences of IVD deficiency.
Jurkat cells are an immortalized human T-cell leukemia line exhibiting CD4+ surface expression, widely utilized to explore T-cell receptor signaling, apoptosis, and immune function. As a suspension cell line with well-characterized growth properties, Jurkat cells provide a robust platform for genetic modification and metabolic studies.
The IVD gene encodes isovaleryl-CoA dehydrogenase, which catalyzes the conversion of isovaleryl-CoA to 3-methylcrotonyl-CoA in mitochondria, representing the third step in the leucine degradation pathway. This reaction requires FAD as a cofactor and electron transfer flavoprotein (ETF) as an electron acceptor, ultimately feeding acetyl-CoA and acetoacetate into the tricarboxylic acid cycle and ketone body production. IVD activity is regulated by leucine availability, mitochondrial import machinery, and the transcription factor PPARalpha, linking amino acid catabolism to cellular energy homeostasis. Disruption of IVD leads to accumulation of isovaleryl-CoA and related organic acids, mirroring the metabolic signature of isovaleric acidemia.
In Jurkat T lymphocytes, knockout of IVD abolishes isovaleryl-CoA dehydrogenase activity, causing accumulation of isovaleryl-CoA and its downstream metabolites. Because T-cell activation and proliferation are energetically demanding and rely on metabolic reprogramming, this polyclonal knockout population offers a unique tool to examine how leucine catabolism intersects with immune cell metabolism. The resulting metabolic stress and potential toxicity from accumulated organic acids can be used to model isovaleric acidemia and other mitochondrial enzyme deficiencies in an immune-relevant cellular environment.
Researchers can employ these polyclonal knockout cells in a variety of downstream applications, including western blotting for IVD confirmation, RT-qPCR analysis of IVD transcript levels, and enzyme activity assays using isovaleryl-CoA as substrate. LC-MS-based metabolite profiling permits quantitation of isovaleryl-CoA accumulation, while cell viability assays under leucine-rich conditions enable assessment of metabolic sensitivity. Metabolic flux analysis and functional complementation experiments expressing wild-type or mutant IVD can further elucidate the role of this enzyme in branched-chain amino acid metabolism. These tools support studies in metabolic disorders, drug screening for metabolic modulators, and investigation of mitochondrial function in T-cell biology. For further technical details or ordering information, please contact Ascent Research.
