The PDHB Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited B lymphocyte population with targeted disruption of the PDHB gene. PDHB encodes the E1 beta subunit of the pyruvate dehydrogenase complex (PDC), which catalyzes pyruvate decarboxylation to acetyl-CoA, linking glycolysis to the TCA cycle. This polyclonal knockout model is designed for investigating metabolic reprogramming when pyruvate oxidation is impaired.
The parental Raji cell line originates from a Burkitt lymphoma, is Epstein-Barr virus (EBV)-positive, and grows in suspension. Widely utilized in immunology and cancer research, Raji cells provide a well-characterized B-cell background for metabolic studies. Their suspension culture facilitates high-throughput assays, and their known genetic profile makes them suitable for metabolic gene knockout experiments.
Within the PDC, PDHB interacts with DLAT (E2), PDHX (E3BP), and DLD (E3) to form the functional complex. Upstream pyruvate dehydrogenase kinases (PDK1?C4) inactivate the complex via phosphorylation, while pyruvate dehydrogenase phosphatases (PDP1?C2) reverse inhibition. Regulatory signals such as insulin, ATP, NADH, and acetyl-CoA control this balance. PDHB disruption eliminates PDC activity, blocking pyruvate conversion to acetyl-CoA and forcing a shift to glutamine anaplerosis to maintain the TCA cycle. This impairs downstream biosynthesis of fatty acids and cholesterol.
In Raji Burkitt lymphoma cells, PDHB knockout exacerbates the metabolic shift seen in the Warburg effect by abolishing glycolytic pyruvate contribution to the TCA cycle. These cells become dependent on alternative fuels, making the model valuable for studying metabolic vulnerabilities in B-cell malignancies. The polyclonal nature ensures population-averaged phenotypes, avoiding clonal bias.
Applications include metabolic flux analysis with Seahorse technology, glucose/lactate assays, and metabolomics to map metabolic rewiring. The model supports research into TCA cycle dysfunction, Warburg effect, lactic acidosis, and immunometabolism. It is also suitable for proliferation, apoptosis, and drug screening studies targeting cancer metabolism. For further information, please contact Ascent Research.
