The ACSF3 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T lymphocyte cell line. This product constitutes a heterogeneous pool of cells carrying targeted disruptions in the ACSF3 gene, achieved through CRISPR/Cas9-mediated gene editing. The polyclonal format enables the study of ACSF3 loss-of-function in a cellular context that retains the inherent diversity of genomic edits, providing a robust model for investigating mitochondrial fatty acid synthesis without the limitations of single-cell clonal selection. These knockout cells are suitable for functional assays requiring stable gene disruption in a leukemic T-cell background.
The Jurkat host cell line was originally established from the peripheral blood of a patient with acute T cell leukemia and serves as a widely used model for T-cell receptor (TCR) signaling and T lymphocyte biology. As an immortalized T-cell lymphoblast line, Jurkat cells exhibit rapid proliferation and are amenable to genetic manipulation, making them an ideal host for generating knockout models. Their well-characterized signaling pathways and metabolic dependencies provide a relevant cellular environment for dissecting the role of mitochondrial metabolic enzymes in immune cell function and leukemogenesis.
ACSF3 encodes a mitochondrial acyl-CoA synthetase that catalyzes the ATP-dependent activation of malonate and methylmalonate to their corresponding coenzyme A esters, namely malonyl-CoA and methylmalonyl-CoA. These products serve as essential substrates for the mitochondrial fatty acid synthesis (mtFAS) pathway, which is critical for the biosynthesis of lipoic acid, a cofactor required by several mitochondrial dehydrogenase complexes. ACSF3 operates within a molecular network involving downstream targets such as MCAT, MECR, and OXSM, and interacts with the acyl carrier protein NDUFAB1. Transcriptional regulation of ACSF3 is influenced by upstream regulators including PPARGC1A, NRF1, and ESRRA, thereby integrating metabolic signals with mitochondrial biogenesis programs. Disruption of ACSF3 function leads to the accumulation of malonic and methylmalonic acid and is associated with the inherited metabolic disorder combined malonic and methylmalonic aciduria.
In Jurkat T lymphocytes, ACSF3 knockout allows the investigation of mtFAS in the context of immune cell metabolism and leukemic transformation. The Jurkat cell line relies on both glycolytic and oxidative metabolism, and perturbations in mitochondrial pathways can influence T-cell activation, proliferation, and apoptosis. This knockout model is particularly valuable for studying how defects in lipoic acid synthesis affect mitochondrial respiration and T-cell metabolic reprogramming, as well as for modeling organic acidurias in a human cell-based system. The polyclonal nature of the knockout population may better recapitulate the heterogeneous metabolic states found in patient samples.
Researchers can employ these ACSF3 knockout Jurkat polyclonal cells in a variety of experimental settings, including metabolite profiling to quantify malonic and methylmalonic acid levels via mass spectrometry, mitochondrial respiration assays using Seahorse technology, and cell proliferation and apoptosis measurements. Further applications include analysis of T-cell activation markers by flow cytometry and Western blotting or RT-qPCR to confirm ACSF3 disruption. These cells provide a platform for exploring the intersection of mitochondrial metabolism and T-cell biology, as well as for screening potential therapeutic interventions for combined malonic and methylmalonic aciduria. For additional technical information, please contact Ascent Research.
