IDH3G Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the Jurkat human T-cell line, designed for loss-of-function studies of the IDH3G gene. This model enables analysis of mitochondrial NAD+-dependent isocitrate dehydrogenase function in an immortalized T-lymphocyte background without clonal isolation, maintaining a heterogeneous knockout pool for robust comparative experiments. Researchers can directly apply this system to dissect IDH3G??s role in TCA cycle regulation and cellular bioenergetics.
Jurkat cells, a human T-cell leukemia line derived from acute lymphoblastic leukemia, are widely used for T-cell signaling, HIV infection, and apoptosis research. These suspension lymphoblasts are amenable to metabolic and stress signaling perturbations, offering a controlled setting to examine how IDH3G disruption impacts T-cell metabolism and survival. Their rapid proliferation supports high-throughput assays exploring metabolic vulnerabilities in leukemic T cells.
IDH3G encodes the gamma subunit of the mitochondrial NAD+-dependent isocitrate dehydrogenase (IDH3) complex, which catalyzes isocitrate conversion to ??-ketoglutarate in the TCA cycle. The IDH3 heterotetramer requires IDH3A and IDH3B subunits for activity; IDH3G is critical for allosteric regulation. Upstream, PGC-1??, NRF1, and TFAM drive IDH3 expression during mitochondrial biogenesis. IDH3-mediated production of ??-ketoglutarate and NADH feeds oxidative phosphorylation, generating ATP. IDH3G knockout disrupts complex formation, reduces TCA flux, and lowers NADH, impairing energy metabolism and redox homeostasis.
In Jurkat leukemic T cells, loss of IDH3G likely impairs NADH and ??-ketoglutarate output, prompting metabolic rewiring toward glycolysis or glutaminolysis. This reprogramming can affect proliferation, apoptosis, and cytokine production, relevant to mitochondrial disorders and cancer metabolism. The model uncovers metabolic dependencies in leukemic T cells, highlighting the intersection of TCA cycle integrity and oncogenic signaling.
Applications include TCA cycle analysis via Seahorse mitochondrial respiration assays, NADH/NAD+ measurement, and LC-MS metabolite profiling. Additional uses encompass immunometabolism, drug sensitivity screens, and metabolic flux analysis, validated by western blotting, RT-qPCR, and flow cytometry. Typical studies examine IDH3G??s impact on ??-ketoglutarate production and T-cell activation or apoptosis. For technical support, contact Ascent Research.
