The AGK Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the AGK gene in the human Jurkat T-lymphocyte line. This polyclonal pool comprises a heterogeneous mix of edited cells, reflecting diverse CRISPR/Cas9-mediated gene disruption events without single-cell cloning. The product provides a reliable loss-of-function model for investigating AGK-dependent mitochondrial lipid metabolism in a consistent T-cell background.
Jurkat cells, an immortalized human T-lymphocyte line derived from the peripheral blood of a 14-year-old boy with T-cell leukemia, are extensively used to study T-cell receptor signaling, activation, and apoptosis. Their well-characterized biology, robust growth, and genetic tractability make them an ideal host for generating knockout models to dissect immune signaling pathways and cell death mechanisms.
AGK encodes a mitochondrial acylglycerol kinase that phosphorylates monoacylglycerol and diacylglycerol to produce lysophosphatidic acid (LPA) and phosphatidic acid (PA), respectively. These lipid second messengers activate LPA receptors (LPAR1?C6) and the mTOR pathway, linking mitochondrial lipid metabolism to cellular signaling. AGK activity is influenced by PPAR signaling, cellular stress, and energy status, and the kinase interacts with TIM22 complex components (TIMM22, TIMM9, TIMM10), implicating it in mitochondrial protein import. Mutations in AGK cause Sengers syndrome, featuring congenital cataracts, hypertrophic cardiomyopathy, and lactic acidosis, highlighting its critical role in mitochondrial function and lipid homeostasis.
In Jurkat T cells, AGK knockout disrupts mitochondrial lipid metabolism, impairing LPA and PA production and consequently attenuating downstream mTOR signaling. This perturbation is expected to alter mitochondrial integrity, apoptotic sensitivity, and T-cell receptor-driven responses, given Jurkat cells?? dependence on mitochondrial function for survival and activation. The knockout model thus enables precise dissection of how AGK-mediated lipid metabolism influences T-cell apoptosis and immune signaling.
This polyclonal knockout cell pool is applicable to diverse experimental workflows, including mitochondrial lipid metabolism studies, T-cell apoptosis research, and drug screening for Sengers syndrome or cancer metabolism. Typical assays include Western blotting for cleaved caspase-3 and PARP, flow cytometric measurement of mitochondrial membrane potential (e.g., TMRE staining), RT-qPCR analysis of lipid metabolic genes, and lipidomics profiling to quantify LPA and PA species. Cell viability and T-cell activation assays further enable functional characterization. For technical inquiries, please contact Ascent Research.
