The GOT1 Knouckout Jurkat Polyclonal Cells are a polyclonal population of Jurkat cells with CRISPR/Cas9-mediated disruption of the GOT1 gene, encoding glutamate-oxaloacetate transaminase 1. This model offers a heterogeneous loss-of-function system to study GOT1-dependent metabolism in a T-cell leukemia context.
The Jurkat cell line, an immortalized human CD4+ T lymphocyte line derived from a 14-year-old male with T cell leukemia, is widely employed in T-cell signaling, apoptosis, and cancer metabolism research. Its leukemic origin and rapid proliferation make it ideal for examining metabolic adaptations in malignant T cells.
GOT1 catalyzes the reversible transamination of aspartate and ??-ketoglutarate to oxaloacetate and glutamate within the malate-aspartate shuttle, regenerating cytosolic NAD+ for glycolysis and redox balance. This enzyme interconnects with aspartate, glutamate, and arginine biosynthesis pathways in central carbon metabolism. Upstream regulators include MYC and HIF1A, along with nutrient deprivation, while downstream outputs encompass aspartate, glutamate, and support for nucleotide and glutathione synthesis. GOT1 partners with MDH1, GOT2, and citrate synthase to coordinate mitochondrial-cytosolic reducing equivalent transfer. Loss of GOT1 disrupts these interactions, impairing NAD+ regeneration and aspartate supply, which alters redox homeostasis and biosynthetic capacity.
In Jurkat cells, GOT1 is critical for metabolic fitness and proliferation, as T lymphocytes depend on the malate-aspartate shuttle to sustain glycolytic flux under high metabolic demand. CRISPR/Cas9-mediated GOT1 disruption in this polyclonal population reduces NAD+ regeneration and aspartate production, compromising nucleotide biosynthesis and glutathione synthesis, thereby altering redox balance and impairing proliferation. This model captures metabolic vulnerabilities of leukemia cells and is valuable for investigating amino acid and glucose adaptation. The polyclonal nature allows study of metabolic heterogeneity, supporting drug target validation and metabolic reprogramming research.
These cells are suited for assays including Western blotting, RT-qPCR, LC-MS metabolomics, and Seahorse flux analysis to probe metabolic changes. Functional assays such as proliferation, apoptosis, flow cytometry, and nucleotide quantification further characterize GOT1-dependent effects. This model is particularly useful for malate-aspartate shuttle studies in leukemia, T-cell metabolic reprogramming, and validating GOT1 as a therapeutic target in pancreatic cancer, breast cancer, and liver dysfunction. For further information, please contact Ascent Research.
