The BDH1 Knockout Jurkat Polyclonal Cells are a population of Jurkat T lymphocytes with CRISPR/Cas9-mediated disruption of the BDH1 gene, generating a heterogeneous pool of loss-of-function mutants. This model ablates the mitochondrial enzyme D-3-hydroxybutyrate dehydrogenase 1, enabling detailed investigation of ketone body metabolism in a human T-cell leukemia background. The polyclonal format avoids clonal biases and is suited for studying metabolic vulnerabilities and ketolysis in immune cells.
Jurkat cells are an immortalized human T lymphocyte line derived from acute T-cell leukemia, widely used for studying T-cell signaling, immunology, and leukemia. These suspension lymphoblasts exhibit high glycolytic activity, making them an excellent platform for exploring alternative fuel utilization such as ketone bodies. The BDH1 knockout in Jurkat cells offers a focused context for evaluating the role of ketolysis in leukemic T-cell metabolism and survival under nutrient stress.
BDH1 encodes a mitochondrial matrix enzyme that catalyzes NAD+-dependent oxidation of D-3-hydroxybutyrate to acetoacetate, generating NADH and feeding the TCA cycle. This key step in ketolysis is transcriptionally regulated by PPAR?? and FOXA2 in response to fasting/ketogenic diet, and modulated by the deacetylase SIRT3, which also interacts with BDH1. The enzyme operates within a metabolic network including HMGCS2, HMGCL, OXCT1, and ACAT1. In Jurkat cells, BDH1 disruption impairs acetoacetate production, redox balance, and TCA flux, affecting overall energy metabolism.
Disruption of BDH1 in Jurkat cells is predicted to shift metabolism away from ketone-dependent respiration toward glycolysis and glutaminolysis, recapitulating metabolic rewiring observed in leukemia. This knockout model allows investigation of ketone body utilization in cancer cell proliferation and survival under metabolic stress. Loss of BDH1 may sensitize cells to NAD+-dependent apoptosis or alter responses to ketogenic interventions, while the polyclonal population captures heterogeneous metabolic adaptations and redox homeostasis outcomes.
Applications include studying ketone body metabolism in immune cells, the role of ketolysis in T-cell function and leukemia, and mitochondrial redox regulation. Standard assays include Western blotting (BDH1, OXCT1), RT-qPCR, Seahorse metabolic flux analysis, ketone body utilization, and NAD+/NADH ratio measurements. The model is also used for apoptosis/viability under metabolic stress and RNA-seq profiling of metabolic gene networks. For further details, contact Ascent Research.
