ACAD11 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T lymphocyte line, engineered for disruption of the ACAD11 gene. This product provides a mixed population of edited cells with loss-of-function mutations at the ACAD11 locus, enabling studies of mitochondrial fatty acid metabolism in an immortalized T-cell context. The polyclonal format bypasses clone-specific artifacts, offering a heterogeneous model that more closely reflects physiological variability in gene disruption. As a pooled knockout model, it is particularly suited for experiments requiring population-level metabolic phenotyping, such as metabolic flux analyses or drug sensitivity screens, without the need for single-cell-derived clones.
The parental Jurkat cell line is an immortalized human T lymphocyte derived from an acute T-cell leukemia patient and is widely used to study T-cell receptor (TCR) signaling, activation, and apoptosis. Jurkat cells exhibit robust glycolytic metabolism characteristic of transformed lymphocytes but retain the capacity for oxidative phosphorylation, making them a relevant model for investigating the interplay between mitochondrial function and immune cell biology. Their utility in immunometabolism research stems from their ability to undergo metabolic reprogramming upon stimulation, mirroring key aspects of primary T-cell metabolism. The generation of polyclonal ACAD11 knockout populations in this background thus provides a tool to dissect how fatty acid oxidation contributes to T-cell activation, proliferation, and viability under both basal and stress conditions.
ACAD11 encodes a mitochondrial acyl-CoA dehydrogenase that catalyzes the initial ??,??-dehydrogenation of long-chain acyl-CoA esters during fatty acid ??-oxidation, transferring electrons to electron transfer flavoprotein (ETF) for subsequent oxidative phosphorylation. This reaction supplies reducing equivalents to the mitochondrial electron transport chain, driving ATP synthesis and influencing reactive oxygen species (ROS) production. ACAD11 activity is under transcriptional regulation by factors including PPARA, PPARGC1A (PGC-1??), SIRT1, and ESRRA, which control mitochondrial biogenesis and fatty acid oxidation gene expression. The enzyme interacts with ETF, ETFDH, and other acyl-CoA dehydrogenases such as ACADVL, ACADM, and ACADS, forming part of a larger ??-oxidation complex. Knockout of ACAD11 disrupts conversion of acyl-CoAs to enoyl-CoAs, leading to accumulation of upstream lipid species and reduced electron flux to the respiratory chain, thereby impairing mitochondrial ATP output and altering cellular redox balance.
In T lymphocytes, fatty acid oxidation supports energy homeostasis, particularly during activation and memory cell differentiation, where metabolic reprogramming toward oxidative phosphorylation is critical. Jurkat cells, despite their leukemic origin, retain dependencies on mitochondrial metabolism for survival and proliferation, and ACAD11 disruption imposes a metabolic blockade that can be exploited to study the role of ??-oxidation in T-cell acute lymphoblastic leukemia. The polyclonal knockout population allows assessment of heterogeneous metabolic adaptation, as subclones may exhibit varying degrees of compensatory glycolysis or amino acid catabolism. This model is thus useful for investigating how ACAD11 deficiency alters TCR signaling, proliferation, apoptosis, and mitochondrial membrane potential, and for evaluating the metabolic vulnerabilities of T-cell leukemia in the context of fatty acid oxidation disorders such as ACAD11 deficiency, which is associated with mitochondrial encephalomyopathy and cardiomyopathy.
Typical applications include quantification of mitochondrial respiration and fatty acid oxidation by Seahorse metabolic flux analysis and radiolabeled fatty acid oxidation assays, measurement of ATP levels via luciferase-based assays, and assessment of mitochondrial membrane potential using JC-1 or TMRE dyes. Flow cytometry can be used to monitor cell proliferation, activation markers, and reactive oxygen species, while Western blotting and RNA-seq enable profiling of OXPHOS complex expression and global metabolic gene networks. This polyclonal knockout system is ideal for screening small-molecule modulators that target metabolic pathways in leukemia or for studying the crosstalk between PPAR signaling and T-cell function. For additional information or customized requests, please contact Ascent Research.
