The ECH1 Knockout Jurkat Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T lymphocyte line, designed for loss-of-function studies of enoyl-CoA hydratase 1 (ECH1). This polyclonal pool provides a heterogeneous genetic background, reflecting population-level disruption of ECH1 while preserving overall cellular diversity for robust functional assays in mitochondrial fatty acid metabolism.
Jurkat cells are an immortalized T lymphocyte line established from the peripheral blood of a 14-year-old male with acute T cell leukemia (ATL). Exhibiting a mature T cell phenotype, Jurkat cells are a cornerstone model for T cell receptor signaling, apoptosis, and immune function. Their malignant origin additionally renders them valuable for probing metabolic rewiring in leukemia, particularly the role of lipid utilization in cancer cell energetics.
ECH1 functions as a mitochondrial enoyl-CoA hydratase and isomerase, catalyzing the isomerization of 3-cis-enoyl-CoA and 3-trans-enoyl-CoA to trans-2-enoyl-CoA, a critical step in the ??-oxidation of unsaturated fatty acids. This reaction occurs downstream of the carnitine palmitoyltransferase system (CPT1, CACT, CPT2) and acyl-CoA dehydrogenases, with ECH1 forming functional associations with other mitochondrial hydratases, including ECHS1 and the HADHA/HADHB heterotrimer. ECH1 expression is transcriptionally controlled by PPAR??, PPAR??, and PGC-1??, and its enzymatic activity feeds forward to generate acetyl-CoA, NADH, FADH2, and ATP, linking fatty acid degradation to energy production. Disruption of ECH1 precipitates the build-up of incompletely metabolized fatty acid intermediates and attenuates oxidative phosphorylation capacity.
In T lymphocytes, fatty acid oxidation (FAO) is increasingly recognized as a pivotal metabolic program supporting activation, proliferation, and effector function, with profound implications for immunoregulation and leukemogenesis. The ECH1 knockout in Jurkat cells thus provides an incisive tool for dissecting how mitochondrial unsaturated FAO intersects with T cell bioenergetics, signaling dynamics, and apoptotic susceptibility. Within the context of ATL-derived cells, this model further permits the dissection of FAO-dependent survival mechanisms and metabolic susceptibilities that may be therapeutically exploitable.
This polyclonal knockout product enables a wide spectrum of experimental applications, including high-resolution metabolic flux analyses using Seahorse fatty acid oxidation stress tests, 14C-palmitate oxidation kinetics, and ATP bioluminescence assays. Complementary phenotypic assessments include Oil Red O staining for neutral lipid accumulation, flow cytometry for mitochondrial mass quantitation, and Annexin V-based apoptosis profiling. The cells are also suited for RT-qPCR and Western blotting validation of pathway components, drug library screening for lipid metabolism modulators, and investigations into metabolic reprogramming in hematologic malignancies. For additional details, please contact Ascent Research.
