The DLAT Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T lymphocyte leukemia cell line. This product comprises a heterogeneous pool of cells carrying disruptions in the DLAT gene, which encodes the E2 subunit (dihydrolipoamide acetyltransferase) of the pyruvate dehydrogenase complex (PDC). The polyclonal nature ensures a diverse array of loss-of-function mutations across the cell population, suitable for population-level studies of metabolic gene disruption. The cells are designed as a research tool for investigating the functional consequences of DLAT ablation in a T cell context.
Jurkat cells are an immortalized T lymphocyte line established from the peripheral blood of a 14-year-old male with acute T cell leukemia. They serve as a widely used model system for studying T cell receptor (TCR) signaling, activation, apoptosis, and leukemogenesis. Their rapid growth, suspension culture characteristics, and well-characterized signaling pathways make them particularly amenable to genetic manipulation and phenotypic analysis. The Jurkat background provides a relevant cellular environment for exploring metabolic regulation within T cells, especially under conditions that mimic leukemic transformation.
DLAT encodes the E2 core component of the pyruvate dehydrogenase complex, a critical mitochondrial enzyme that converts pyruvate into acetyl-CoA, linking glycolysis to the tricarboxylic acid (TCA) cycle. DLAT is dynamically regulated by upstream kinases (PDK) and phosphatases (PDP), which respond to insulin signaling and metabolic substrates such as pyruvate and NAD+. Within the PDC, DLAT interacts with E1 (pyruvate dehydrogenase) and E3 (dihydrolipoamide dehydrogenase) subunits, as well as E3-binding protein (E3BP), utilizing lipoamide cofactors to transfer acetyl groups. Downstream, DLAT activity drives acetyl-CoA production, NADH generation, and TCA cycle entry, with implications for fatty acid synthesis. Disruption of DLAT therefore forces cells to rely on alternative metabolic pathways, such as glutaminolysis, to sustain mitochondrial function.
In the context of Jurkat T cells, DLAT knockout creates a metabolic vulnerability model relevant to both pyruvate dehydrogenase deficiency and cancer metabolic reprogramming. T lymphocytes undergo dramatic metabolic shifts upon activation, transitioning from oxidative phosphorylation to aerobic glycolysis, and genetic disruption of DLAT can reveal how these metabolic checkpoints regulate proliferation, survival, and effector functions. This model is particularly valuable for dissecting the interplay between glucose oxidation and glutamine dependence in leukemia cells, potentially uncovering targetable metabolic liabilities. Additionally, it provides a platform for studying neurometabolic disorders linked to PDC dysfunction, as T cells can reflect systemic metabolic defects.
Investigators can employ this polyclonal knockout population in a range of assays to monitor metabolic adaptation. Seahorse extracellular flux analysis permits real-time measurement of mitochondrial respiration and glycolysis, elucidating the shift toward glutaminolysis. Western blotting and RT-qPCR confirm DLAT depletion and assess expression changes in interacting partners like PDK or PDP. Metabolomic profiling can quantify alterations in acetyl-CoA, NADH, TCA cycle intermediates, and glutamine-derived metabolites. Cell proliferation and apoptosis assays reveal the functional consequences under nutrient-limited or drug-treated conditions. Flow cytometry with mitochondrial dyes (e.g., MitoTracker) further characterizes mitochondrial fitness. For detailed protocols and technical support, contact Ascent Research.
