ALDH4A1 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of Jurkat T lymphocytes with targeted disruption of the ALDH4A1 gene. This loss-of-function model is designed for investigating proline catabolism and glutamate metabolism in a human immune cell context. The cells are derived from the Jurkat cell line and edited without clonal selection, resulting in a heterogeneous pool that maintains genetic diversity while achieving effective ALDH4A1 suppression, thus supporting pooled functional assays and phenotypic characterization.
The Jurkat host cell line is an immortalized human T lymphocyte line originating from a patient with acute T-cell leukemia. These cells display a lymphoblast-like phenotype and are extensively used as a model system for T cell activation, proliferation, and apoptosis. Jurkat cells are amenable to genetic manipulation and retain key T cell signaling pathways, making them a versatile platform for studying metabolic regulation in lymphocyte biology and leukemia.
ALDH4A1 encodes a mitochondrial NAD+-dependent dehydrogenase that oxidizes glutamic gamma-semialdehyde to glutamate, the final step of proline catabolism. This reaction follows PRODH-mediated conversion of proline to ??1-pyrroline-5-carboxylate (P5C), which spontaneously generates the ALDH4A1 substrate. Resulting glutamate can enter the TCA cycle or be used for glutathione (GSH) synthesis. ALDH4A1 operates as a homodimer, is influenced by substrate availability and potentially PGC-1??, and interconnects proline, arginine, and ornithine metabolism.
In Jurkat T cells, ALDH4A1 disruption abolishes glutamate production from proline, impairing TCA cycle fueling, GSH biosynthesis, and nucleotide precursor generation??processes crucial for T cell activation and proliferation. This knockout model may exhibit reduced cell growth, altered redox status, and heightened sensitivity to metabolic stress, providing insights into the metabolic dependencies of malignant T cells. It also serves as a tool for studying hyperprolinemia type II, a disorder of proline accumulation linked to neurodevelopmental abnormalities, within an immune cell context.
This ALDH4A1 polyclonal knockout product is suitable for diverse biochemical and functional assays, including liquid chromatography-mass spectrometry (LC-MS)-based metabolite profiling to quantify proline and glutamate levels, Seahorse metabolic flux analysis to assess mitochondrial respiration and glycolysis, and cell proliferation (MTT or EdU) and apoptosis (Annexin V) assays. Additional applications include investigating the role of proline catabolism in T cell cancer metabolism, modeling hyperprolinemia type II metabolic signatures, and exploring amino acid deprivation responses. The polyclonal format ensures population-level consistency for bulk experiments. For further information, please contact Ascent Research.
