The EHHADH Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal Jurkat cell population featuring targeted disruption of EHHADH. This knockout pool serves as a loss-of-function model for studying peroxisomal L-bifunctional enzyme activity within a T lymphocyte background, avoiding the need for clonal isolation. The polyclonal nature preserves biological heterogeneity while enabling robust functional genomics and metabolic investigations.
Jurkat cells are an immortalized T lymphocyte line derived from an acute T cell leukemia patient, widely used to model T cell receptor signaling, apoptosis, and leukemogenesis. Their well-characterized signaling networks and dependency on specific metabolic pathways make them an ideal host to investigate how peroxisomal fatty acid oxidation influences T cell activation, proliferation, and survival. This knockout thus allows direct interrogation of lipid metabolism in a malignant T cell context.
The EHHADH gene encodes the peroxisomal L-bifunctional enzyme, responsible for the hydration and dehydrogenation steps of fatty acid beta-oxidation, producing acetyl-CoA and NADH. Its expression is controlled by PPAR??, thyroid hormone, and insulin, and it interacts with PEX5, PEX7, ACOX1, and ACAA1 within a complex that includes SCP2. Disruption of EHHADH severs the peroxisomal beta-oxidation pathway, impairing long-chain fatty acid catabolism and disturbing lipid homeostasis.
In Jurkat T cell leukemia cells, EHHADH knockout creates a metabolic defect that specifically impairs peroxisomal long-chain fatty acid oxidation, likely leading to lipid accumulation and altered energy metabolism. As activated T cells rely on fatty acid oxidation for proliferation and effector functions, and metabolic reprogramming is critical for leukemia survival, this knockout model provides a valuable tool to dissect the role of peroxisomal lipid metabolism in T cell malignancy. Researchers can examine the interplay between peroxisomal and mitochondrial oxidation, redox balance, and sensitivity to metabolic inhibitors.
These polyclonal knockout cells are suited for studying peroxisomal fatty acid oxidation in leukemic T cells, metabolic reprogramming in cancer, and peroxisomal function in immune cells. Compatible assays include fatty acid oxidation flux measurements using labeled palmitate, peroxisomal enzyme activity assays, Oil Red O lipid staining, ATP quantification, and flow cytometry for T cell activation, apoptosis, and proliferation. The model also supports drug screening for peroxisomal disorders or metabolic syndrome. For further details, please contact Ascent Research.
