The HADHB Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout cell population derived from the Jurkat T-cell acute lymphoblastic leukemia (T-ALL) line. This product provides a heterogeneous pool of Jurkat cells with targeted disruption of the HADHB gene, establishing a loss-of-function model for the mitochondrial trifunctional protein ??-subunit. The polyclonal format preserves population-level diversity, enabling robust analysis of metabolic adaptations without clonal selection bias.
The Jurkat host line (clone E6-1) was originally established from the peripheral blood of a 14-year-old male with acute T-cell leukemia. These cells display a mature T-lymphocyte phenotype and serve as a widely accepted model for T-ALL, characterized by aberrant proliferative signaling and dependency on specific metabolic pathways. The Jurkat background offers a physiologically relevant context for studying how HADHB loss impacts cancer cell bioenergetics.
HADHB encodes the ??-subunit of the mitochondrial trifunctional protein (MTP), which catalyzes the final step of long-chain fatty acid ??-oxidation??specifically, the thiolytic cleavage of 3-ketoacyl-CoA into acetyl-CoA. The MTP ??-subunit (HADHA) forms a stoichiometric complex with HADHB, and together they operate downstream of the carnitine shuttle (CPT1A, CACT, CPT2) and very-long-chain acyl-CoA dehydrogenase (ACADVL). HADHB activity is transcriptionally regulated by PPARA and its coactivator PPARGC1A, as well as by the bile acid receptor NR1H4. Functional interaction partners include ACADVL, EHHADH, and HSD17B10. Through its thiolase activity, HADHB directly governs the production of acetyl-CoA, NADH, and FADH2, thereby coupling fatty acid catabolism to the electron transport chain and ATP synthesis. Disruption of HADHB abrogates this thiolase step, leading to accumulation of long-chain acyl-CoA intermediates and a reduction in mitochondrial acetyl-CoA and ATP output.
In the Jurkat T-ALL context, HADHB knockout forces a metabolic shift away from fatty acid-dependent oxidative phosphorylation, engendering increased reliance on glucose and glutamine for energy and anabolic precursors. This re-wiring creates a model of mitochondrial energy insufficiency that can be exploited to study cancer cell metabolic plasticity, vulnerability to nutrient deprivation, and sensitivity to inhibitors of alternative fuel pathways. The polyclonal nature allows the capture of diverse clonal responses, mirroring the heterogeneity of metabolic adaptations seen in leukemia.
This polyclonal knockout cell population is suited for applications such as dissecting fatty acid oxidation deficiencies in cancer, evaluating mitochondrial dysfunction, and screening metabolic inhibitors in T-ALL. Representative assays include [14C]-palmitate oxidation tracing, Seahorse XF Mito Stress Tests, immunoblotting for HADHB and associated subunits, NADH/NAD+ ratio measurements, ATP luminescence assays, and proliferation analyses under lipid-rich or glucose-depleted conditions. Acyl-carnitine profiling by LC-MS/MS further enables interrogation of intermediate accumulation. For additional information or technical assistance, please contact Ascent Research.
