The ACOT2 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T lymphocyte cell line, engineered to disrupt the ACOT2 gene. This polyclonal pool consists of a heterogeneous mixture of edited cells harboring targeted disruptions at the ACOT2 locus, providing a loss-of-function model for studying ACOT2 function in acyl-CoA metabolism within a T cell leukemia background.
The host Jurkat cell line, derived from a 14-year-old male with acute T cell leukemia, is a well-established model for T cell signaling, apoptosis, and immunometabolism. Jurkat cells exhibit rapid proliferation and defined signaling cascades, making them suitable for metabolic studies. Their reliance on both glycolysis and fatty acid oxidation for energy and biosynthesis underscores their utility in dissecting lipid metabolic pathways relevant to leukemogenesis and T cell activation.
ACOT2 hydrolyzes long-chain acyl-CoA thioesters, such as palmitoyl-CoA and oleoyl-CoA, to free fatty acids and CoA, thereby regulating intracellular fatty acid pools and mitochondrial ??-oxidation. ACOT2 expression is regulated upstream by PPAR?? and AMPK signaling in response to nutritional cues. The released free fatty acids and CoA serve as substrates and cofactors for ??-oxidation, influencing CPT1 activity and acetyl-CoA carboxylase-mediated lipid synthesis. Disruption of ACOT2 alters acyl-CoA homeostasis, potentially shifting the balance between lipid oxidation and storage and impacting mitochondrial ??-oxidation flux and lipid-mediated signaling.
In Jurkat cells, ACOT2 knockout is expected to disrupt acyl-CoA metabolism, affecting fatty acid oxidation and lipid availability crucial for T cell proliferation, signaling, and survival. This model permits exploration of how leukemic cells rewire lipid metabolism to support growth, and how ACOT2 influences immunometabolic phenotypes. The polyclonal knockout population captures a spectrum of editing efficiencies, reflecting tumor heterogeneity, and provides a versatile platform for studying metabolic vulnerabilities in T cell leukemia.
Researchers can employ this knockout model in assays such as Western blotting and RT-qPCR for confirmation of gene disruption, Seahorse-based fatty acid oxidation assays, metabolic flux analysis, and lipidomic profiling to assess lipidome remodeling. Flow cytometry enables measurement of proliferation and apoptosis. Applications include dissecting lipid metabolism in T cells, examining fatty acid oxidation in leukemia, studying metabolic reprogramming in cancer, and functionally analyzing ACOT2 in immune cell function. For further details or technical assistance, please contact Ascent Research.
