The ACADM Knouckout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ACADM gene in human Jurkat T lymphocytes. This loss-of-function model disrupts medium-chain acyl-CoA dehydrogenase (MCAD), the enzyme initiating mitochondrial medium-chain fatty acid beta-oxidation. The polyclonal pool ensures heterogeneous gene disruption, suitable for studying ACADM deficiency without clonal selection bias, offering flexibility for metabolic and cancer research applications.
Jurkat is an immortalized human T-cell leukemia line widely used to study T-cell receptor signaling, apoptosis, and leukemogenesis. Derived from an acute T-cell leukemia patient, Jurkat cells grow in suspension and are amenable to high-throughput assays, providing a reliable host for interrogating metabolic pathways. Its robust proliferation and well-characterized signaling networks make it ideal for metabolic studies.
ACADM encodes MCAD, a mitochondrial flavoenzyme catalyzing the first dehydrogenation of medium-chain acyl-CoAs. It functions within the beta-oxidation pathway, requiring electron transfer flavoprotein (ETF) and ETF-ubiquinone oxidoreductase. Transcriptional activation by PPAR-alpha/RXR-alpha and coactivator PGC-1alpha, downstream of glucagon/cAMP signaling, drives expression. MCAD produces acetyl-CoA and NADH, feeding the TCA cycle and oxidative phosphorylation. Disruption thus blocks medium-chain fatty acid utilization, causing acyl-carnitine accumulation and reduced mitochondrial ATP production, sensitizing cells to energy stress.
In Jurkat cells, ACADM knockout creates a model of medium-chain acyl-CoA dehydrogenase deficiency (MCADD) and fatty acid oxidation disorders. T lymphocytes, though glycolytic, depend on fatty acid oxidation during nutrient limitation or stress. Loss of MCAD impairs this metabolic flexibility, altering apoptosis and survival pathways. This system permits analysis of how mitochondrial fatty acid metabolism defects affect leukemic T-cell proliferation, drug sensitivity, and immune signaling.
Applications include metabolic disorder modeling, MCADD research, fatty acid oxidation analysis, mitochondrial dysfunction studies, and drug screening. Typical assays: Western blot/RT-qPCR for ACADM, fatty acid oxidation assay, Seahorse metabolic flux analysis, cell viability under metabolic stress, flow cytometry for mitochondrial membrane potential, and acyl-carnitine metabolomics. For more information, contact Ascent Research.
