The ACADVL Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the Jurkat T lymphocytic line, offering a loss-of-function model for targeted disruption of the ACADVL gene. This heterogeneous pool enables robust population-level studies of very long-chain acyl-CoA dehydrogenase (VLCAD) function without the biases of clonal selection, while retaining native T-cell signaling and metabolic properties. The polyclonal format provides a versatile tool for investigating gene function in a physiologically relevant context.
The Jurkat host cell line originates from an acute T-cell leukemia patient and is widely used for T-cell receptor signaling, apoptosis, and leukemia research. These cells exhibit metabolic plasticity, shifting between glycolysis and oxidative phosphorylation, making them particularly suitable for studying mitochondrial long-chain fatty acid ??-oxidation and the consequences of ACADVL deficiency in a T-cell environment.
ACADVL catalyzes the initial and rate-limiting step of mitochondrial long-chain fatty acid ??-oxidation, mediating the ??,??-dehydrogenation of long-chain acyl-CoA to enoyl-CoA while transferring electrons to electron transfer flavoprotein (ETF). This reaction is transcriptionally regulated by PPAR??, a nuclear receptor activated by fasting and AMPK signaling, and is essential for generating acetyl-CoA, NADH, and FADH2 to drive ATP production. The enzyme functions in concert with ETF and ETF-ubiquinone oxidoreductase, and its activity depends on upstream components including carnitine palmitoyltransferase 1 (CPT1) and acyl-CoA synthetase. Disruption of ACADVL thus blocks electron flux, impairs fatty acid oxidation, and leads to accumulation of long-chain acylcarnitines.
In Jurkat cells, ACADVL knockout recapitulates key metabolic features of VLCAD deficiency, a disorder associated with cardiomyopathy, hypoglycemia, and sudden infant death. Loss of VLCAD forces T cells to rely more heavily on glucose and glutamine for energy, creating a model to dissect metabolic reprogramming during T-cell activation, differentiation, or leukemic transformation. This system also enables investigation of how impaired fatty acid oxidation affects T-cell receptor signaling and PPAR?? regulatory networks, providing insight into the metabolic vulnerabilities of leukemic cells.
Typical research applications include acylcarnitine profiling by tandem mass spectrometry, fatty acid oxidation flux assays using radiolabeled palmitate, and ATP measurement to quantify metabolic deficits. Mitochondrial respiration is assessed via Seahorse analysis under fatty acid load, with molecular validation by western blot for ACADVL and RT-qPCR for expression changes. Additional phenotypic assays, such as cell viability under fatty acid supplementation, apoptosis detection, and PPAR?? activation reporter systems, further characterize the knockout effects. This polyclonal model is a valuable resource for drug screening in VLCAD deficiency, studying T-cell metabolic signaling, and exploring mitochondrial biology. For further information, please contact Ascent Research.
