The ACADSB Knockout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population harboring targeted disruption of the ACADSB gene. This loss-of-function model is generated on the Jurkat host cell background and provides researchers with a genetically defined tool to interrogate the role of short/branched-chain acyl-CoA dehydrogenase in T-lymphocyte biology. By eliminating ACADSB function, these cells enable systematic investigation of branched-chain amino acid (BCAA) catabolism, mitochondrial fatty acid oxidation, and related metabolic networks in an acute T-cell leukemia context. The polyclonal format ensures a heterogeneous knockout population suitable for pooled functional screens and population-level analyses without the clonal artifacts associated with single-cell-derived lines.
The Jurkat cell line is an immortalized human T lymphoblastoid line originally derived from a patient with acute T-cell leukemia. Widely employed in immunology and cancer research, Jurkat cells serve as a classical model for T-cell receptor signaling, apoptosis, and immune response mechanisms. Their well-characterized signaling pathways and ease of genetic manipulation make them an ideal host for exploring metabolic dependency in T-cell malignancies. The acute leukemia origin also offers a disease-relevant backdrop for assessing how metabolic disruptions impact cell survival, proliferation, and stress responses.
ACADSB encodes a mitochondrial homotetrameric enzyme that catalyzes the ??,??-dehydrogenation of short/branched-chain fatty acyl-CoA thioesters, including isobutyryl-CoA and isovaleryl-CoA, using FAD as a cofactor. This reaction represents a critical step in the degradation of valine and leucine, transferring electrons to electron transfer flavoprotein (ETF) and subsequently to the ETF dehydrogenase system to fuel oxidative phosphorylation. ACADSB activity is regulated by substrate availability, and its expression is transcriptionally controlled by PPARGC1A (PGC-1alpha) in response to metabolic demands such as fasting or high-protein intake. Downstream products include propionyl-CoA, acetyl-CoA, methylacrylyl-CoA, and TCA cycle intermediates, linking BCAA catabolism to central carbon metabolism and ketogenesis. Disruption of ACADSB therefore impedes the conversion of valine-derived isobutyryl-CoA to methacrylyl-CoA and leucine-derived isovaleryl-CoA to 3-methylcrotonyl-CoA, causing a metabolic blockade that can be probed via acylcarnitine profiling.
In the Jurkat T-lymphocyte context, ACADSB knockout holds particular significance due to the emerging role of BCAA metabolism in immune cell function and leukemia cell survival. T cells rely on metabolic reprogramming to support activation, proliferation, and effector functions, and mitochondrial oxidation of BCAAs may contribute to energy homeostasis and biosynthesis. The ACADSB-deficient model allows researchers to dissect how loss of this enzyme alters mitochondrial respiration, nutrient stress responses, and apoptotic signaling in acute leukemia cells. Given the metabolic vulnerabilities often observed in cancer, this system provides a platform to identify dependencies that could be exploited therapeutically in T-cell malignancies.
These polyclonal knockout cells are applicable to a broad range of research areas, including metabolic reprogramming in cancer, immunometabolism, mitochondrial dysfunction, and BCAA-related disorders such as 2-methylbutyrylglycinuria. Researchers can employ a variety of functional assays, including Western blotting for ACADSB, acylcarnitine profiling by mass spectrometry, Seahorse metabolic flux analysis, cell viability measurements under nutrient deprivation (e.g., BCAA withdrawal), RT-qPCR for metabolic gene expression, and apoptosis assays. By combining genetic perturbation with detailed metabolic phenotyping, these cells facilitate mechanistic studies that connect ACADSB activity to T-cell biology and leukemia pathogenesis. For detailed technical support or custom requests, please contact Ascent Research.
