The ETFDH Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from Raji B lymphocytes, designed to disrupt the ETFDH gene encoding mitochondrial electron transfer flavoprotein dehydrogenase. This mixed cell pool preserves the inherent heterogeneity of the parental line while eliminating target gene function, providing a robust loss-of-function model for metabolic studies without clonal selection bottlenecks.
Raji cells are a suspension-adapted human B lymphocyte line established from an EBV-positive Burkitt lymphoma. They maintain key B-cell attributes including antibody secretion and antigen presentation, making them a physiologically relevant platform for investigating humoral immunity and metabolic adaptation in transformed lymphocytes. Their rapid growth and well-characterized signaling landscape facilitate reproducible mitochondrial research.
ETFDH localizes to the mitochondrial inner membrane, where it oxidizes reduced ETF (ETFA/ETFB) and reduces ubiquinone, coupling fatty acid ??-oxidation and branched-chain amino acid catabolism to the respiratory chain at complex III. The flavoprotein requires FAD, derived from riboflavin metabolism, and is regulated by upstream substrates including reduced ETF, fatty acyl-CoA species, and branched-chain ??-keto acids. Downstream, it influences ubiquinone, mitochondrial membrane potential, ATP synthesis, and complex III activity. Disruption of ETFDH uncouples lipid and amino acid oxidation from oxidative phosphorylation, modeling the metabolic blockade observed in glutaric acidemia type II and multiple acyl-CoA dehydrogenase deficiency.
In Raji cells, which reprogram metabolism to sustain proliferation, ETFDH knockout reveals the contribution of fatty acid and amino acid oxidation to energy homeostasis and survival of malignant B cells. Since EBV-positive lymphocytes exhibit altered metabolic profiles, this model helps dissect mitochondrial vulnerabilities in lymphoma. The system also permits testing of riboflavin responsiveness, as some MADD patients benefit from FAD precursor supplementation.
Typical applications include Seahorse metabolic flux analysis to quantify fatty acid oxidation and oxygen consumption, ATP luminescence assays, TMRM/JC-1 mitochondrial membrane potential flow cytometry, and molecular validation by Western blotting and RT-qPCR. These cells serve as a disease model for lipid storage myopathy and glutaric acidemia type II, a platform for B-cell lymphoma metabolic reprogramming studies, and a screening tool for ETFDH deficiency therapies. For further information, please contact Ascent Research.
