The CBR4 Knockout Jurkat Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population targeting the CBR4 gene in the Jurkat T lymphoblastoid cell line. This heterogeneous knockout pool ensures efficient gene disruption without clonal selection, enabling robust loss-of-function experiments while maintaining population diversity. It serves as a versatile tool for studying the metabolic functions of CBR4 in a T cell context.
Jurkat cells are a suspension human T lymphocyte line derived from acute T cell leukemia, widely employed as a model for T cell activation, signal transduction, and leukemia biology. Their well-characterized signaling pathways and ease of genetic manipulation make them an ideal host for CRISPR/Cas9-mediated gene disruption, particularly for investigating metabolic regulation in immune and cancer cells.
CBR4 (MECR) is a mitochondrial enoyl-CoA reductase in the mitochondrial fatty acid synthesis (mtFAS) pathway, where it acts on acyl-ACP substrates downstream of ACSF3, MCAT, and OXSM. CBR4 activity is essential for generating the octanoyl-ACP precursor for lipoic acid biosynthesis by LIAS. Lipoic acid is then transferred by LIPT1 to key dehydrogenase complexes, including the pyruvate dehydrogenase complex (PDHA1, PDHB, DLAT, DLD) and ??-ketoglutarate dehydrogenase complex (OGDH, DLST, DLD), enabling their TCA cycle function. Expression of CBR4 is regulated by NRF1 and PPARGC1A, linking mtFAS to mitochondrial biogenesis and cellular energy status.
Disruption of CBR4 in Jurkat cells impairs mtFAS, leading to defective lipoic acid synthesis and insufficient lipoylation of PDHC and KGDHC. This results in reduced TCA cycle flux, compromised mitochondrial respiration, and altered cellular energy and redox homeostasis. These metabolic changes can affect T cell activation, proliferation, and leukemic growth, making this model valuable for studying mitochondrial dysfunction in immune cell disorders and cancer metabolism.
Researchers can employ this knockout model to investigate the role of mtFAS and lipoic acid metabolism in T cell biology, including mitochondrial dysfunction in immune activation and leukemia. Typical applications include Western blot for lipoylated proteins, Seahorse respirometry, LC-MS-based TCA cycle metabolite profiling, cell proliferation and viability assays, and flow cytometry for mitochondrial mass and membrane potential. RT-qPCR can quantify metabolic gene expression changes, while lipoic acid supplementation rescue experiments validate specific metabolic defects. For further information, please contact Ascent Research.
