The GPT2 Knockout Jurkat Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal population of Jurkat cells with targeted disruption of the GPT2 gene. This polyclonal knockout format generates a diverse allelic pool, enabling investigation of GPT2 loss-of-function without the need for single-cell cloning. The cells are suitable for studying metabolic reprogramming and T-cell biology where ablation of GPT2-mediated alanine transamination is desired.
The host cell line, Jurkat, is an immortalized human T-cell leukemia line derived from the peripheral blood of a male with acute lymphoblastic leukemia. Jurkat cells are widely employed as a model for T-lymphocyte signaling, cytokine production, and activation-induced responses. Their robust growth characteristics and well-characterized signaling pathways make them a preferred system for immunological and metabolic studies.
GPT2 encodes the mitochondrial alanine transaminase, a pyridoxal-phosphate-dependent enzyme that catalyzes the reversible conversion of alanine and ??-ketoglutarate to pyruvate and glutamate. This reaction links amino acid metabolism to the TCA cycle and gluconeogenesis. GPT2 activity is regulated by amino acid availability and transcriptional coactivators including PGC-1alpha and PPARgamma, positioning it at the intersection of nutrient sensing and energy homeostasis. The enzyme functions within the glucose-alanine cycle and directly impacts pyruvate and glutamate pools, influencing anaplerotic flux.
In Jurkat T cells, metabolic reprogramming is critical for activation and effector function. By disrupting GPT2, these knockout cells impair the conversion of alanine to pyruvate, potentially leading to reduced TCA cycle intermediates and compromised energy metabolism. This model is particularly relevant for dissecting the role of mitochondrial amino acid metabolism in T-cell activation, proliferation, and cytokine production, as well as for exploring cancer cell metabolic dependencies.
Researchers can utilize these GPT2 knockout Jurkat cells in a broad range of assays, including Western blotting for GPT2 expression, RT-qPCR, enzyme activity measurements, amino acid profiling, and metabolic flux analysis. The cells are valuable for studying T-cell metabolism, cancer metabolism, neurodegenerative disease pathways, and drug screening targeting mitochondrial aminotransferases. Additional functional readouts may include cell viability assays, flow cytometry for T-cell markers, and lactate/pyruvate quantification. For detailed product specifications and technical support, please contact Ascent Research.
