The ACOT9 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat T-cell line, designed for loss-of-function investigation of ACOT9. This genetically heterogeneous model, generated by CRISPR/Cas9-mediated gene disruption, is suitable for studying ACOT9-dependent mitochondrial functions and their role in T-cell biology and leukemia.
The Jurkat host cell line is an immortalized human T lymphocyte line from an acute lymphoblastic leukemia patient (clone E6-1), widely used to model TCR signaling, apoptosis, and T-cell activation. Its robust signaling pathways and compatibility with metabolic and flow cytometric assays make it an ideal platform for examining the intersection of metabolism and immune function.
ACOT9 encodes a mitochondrial acyl-CoA thioesterase that catalyzes the hydrolysis of medium- to long-chain fatty acyl-CoAs, releasing free fatty acids and CoA within the mitochondrial matrix. This enzymatic activity regulates substrate availability for ??-oxidation and influences mitochondrial energy metabolism. ACOT9 expression is transcriptionally upregulated by PPAR?? and SREBP1, while its functional modulation occurs via mTORC1 and AMPK signaling cascades that are activated downstream of TCR engagement and CD28 co-stimulation. The enzyme physically interacts with ACSL family members (e.g., ACSL1), CPT1A, and mitochondrial trifunctional protein, linking it directly to fatty acid activation, transport, and oxidation machinery. Key downstream effects include alterations in mitochondrial ??-oxidation, ATP synthesis, and ROS generation, with pathway crosstalk involving ACADM, HADHA, PPARA, RXRA, MTOR, and RPS6KB1.
In Jurkat cells, ACOT9 disruption may perturb fatty acid catabolism, alter mitochondrial respiration, and affect acetyl-CoA and ROS homeostasis, potentially impacting T-cell activation, proliferation, and apoptosis. As leukemic cells often depend on mitochondrial metabolism, this polyclonal knockout model enables dissection of ACOT9??s role in leukemia cell survival and metabolic adaptations, providing insights into tumor heterogeneity.
This knockout model is well-suited for investigating the role of mitochondrial acyl-CoA metabolism in T-cell immunometabolism, screening chemical libraries for modulators of fatty acid oxidation, and validating ACOT9 as a potential drug target in leukemia. Researchers can monitor knockout efficiency via Western blot and RT-qPCR, assess metabolic function using radiolabeled palmitate oxidation assays and Seahorse extracellular flux analysis, and quantify mitochondrial mass with MitoTracker. Functional consequences on T-cell biology can be examined through flow cytometry-based apoptosis detection (Annexin V), ROS measurement (DCFDA), and T-cell activation assays (CD69 expression). Cell viability can be evaluated using MTT or ATP luminescence. For further details, contact Ascent Research.
