The ACAD8 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the ACAD8 gene is disrupted in Jurkat human T lymphocytes. This loss-of-function model employs a heterogeneous pool of edited cells, avoiding monoclonal artifacts, to study the isobutyryl-CoA dehydrogenase enzyme. The product facilitates investigation of valine catabolic pathways and mitochondrial metabolism in an immortalized T-cell system, supporting reproducible in vitro assays. The heterogeneous nature of the polyclonal population ensures representation of various editing outcomes, enabling robust statistical comparisons in functional studies.
Jurkat cells, an immortalized T lymphocyte line derived from acute T cell leukemia, are extensively used to model T-cell receptor signaling and apoptosis. Their stable growth properties and well-characterized signaling networks make them ideal for integrating CRISPR/Cas9-mediated gene disruptions in metabolic research. In this ACAD8 knockout context, the leukemic T-cell background enables exploration of branched-chain amino acid catabolism and its impact on energy homeostasis and proliferation, relevant to both normal T-cell biology and leukemia metabolism. This model thus bridges the gap between inborn metabolic errors and cancer cell metabolism.
ACAD8 encodes a mitochondrial FAD-dependent dehydrogenase that catalyzes the oxidation of isobutyryl-CoA to methylacrylyl-CoA in valine catabolism. The enzyme is regulated by metabolic sensors such as AMPK and PPAR family transcription factors PPARA and PPARG, coactivated by PPARGC1A. It functions upstream of HIBCH and ECHS1, ultimately feeding propionyl-CoA and succinyl-CoA into the TCA cycle, and requires electron transfer via ETFA, ETFB, and ETFDH. ACAD8 disruption leads to accumulation of isobutyryl-CoA and isobutyrylglycine, impairing mitochondrial energy metabolism and disrupting cellular redox homeostasis.
In Jurkat cells, ACAD8 knockout recapitulates metabolic features of isobutyryl-CoA dehydrogenase deficiency, an inborn error of valine metabolism leading to organic aciduria. The model enables study of how defective valine catabolism affects mitochondrial function, energy production, and cellular homeostasis in T lymphocytes. Moreover, the leukemic origin of Jurkat cells provides insights into the role of branched-chain amino acid metabolism in cancer cell survival and metabolic reprogramming, highlighting mitochondrial vulnerabilities in leukemia. This dual relevance makes the model a versatile platform for both rare disease research and oncology.
This polyclonal knockout product supports mechanistic studies of valine metabolism, disease modeling for organic acidurias, and drug screening for mitochondrial dysfunction. Researchers can confirm knockout via Western blot, RT-qPCR, and genomic DNA sequencing. Metabolic profiling by LC-MS/MS quantifies acylcarnitines and isobutyrylglycine, while Seahorse flux analysis and flow cytometry assess mitochondrial function and membrane potential, complemented by cell viability assays. These tools enable investigation of T-cell metabolism in leukemia and evaluation of therapeutic candidates. Comprehensive functional assessments are thus supported across multiple experimental endpoints. For further information, contact Ascent Research.
