The ETFB Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population derived from the human Raji B lymphocyte line, engineered for loss of ETFB function. This heterogeneous pool of edited alleles ensures robust representation of genetic perturbations across the population, making it well-suited for bulk functional assays in metabolism and immunology without the need for single-cell cloning. The polyclonal format is ideal for applications where population-level phenotypes are primary readouts.
The parental Raji cell line is an EBV-positive Burkitt lymphoma-derived B lymphocyte model extensively used to investigate humoral immunity, B cell receptor (BCR) signaling, and antigen presentation. Its malignant origin also renders it a relevant system for studying metabolic reprogramming in lymphoma and B-cell biology.
ETFB encodes the beta subunit of electron transfer flavoprotein (ETF), forming a heterodimer with ETFA to shuttle electrons from primary dehydrogenases to ETFDH. Upstream regulators include PPAR-alpha, PGC-1alpha, ERR??, and AMPK signaling. ETF accepts electrons from acyl-CoA dehydrogenases during fatty acid ??-oxidation and from glutaryl-CoA and isovaleryl-CoA dehydrogenases during amino acid catabolism. Electrons are transferred via ETFDH to ubiquinone, feeding into respiratory complex III. This electron relay is vital for ATP synthesis, NAD+/NADH balance, and ROS control. ETFB interacts with ETFA, ETFDH, and dehydrogenases. Its disruption uncouples oxidation from energy production.
In the Raji context, ETFB knockout impairs mitochondrial electron flow and oxidative capacity, potentially affecting B-cell functions such as proliferation, class switching, and antigen presentation. Since lymphoma cells often rely on fatty acid oxidation, this model is valuable for dissecting ETF-dependent metabolic dependencies in B-cell malignancies. It also recapitulates features of glutaric acidemia type II (multiple acyl-CoA dehydrogenase deficiency), enabling studies of toxic acylcarnitine accumulation and metabolic crisis. Additionally, it offers a platform to investigate how mitochondrial stress impacts BCR signaling and immune effector functions.
Key applications include modeling glutaric acidemia type II, examining mitochondrial energy metabolism in B cells, and probing lipid utilization in lymphoma. Researchers can employ a range of downstream assays, such as Western blot and RT-qPCR for target validation, Seahorse-based mitochondrial respirometry, fatty acid oxidation flux analysis, ATP quantification, ROS detection, apoptosis assays under metabolic stress, and targeted metabolomics for acylcarnitine profiles. For additional technical information or custom requests, please contact Ascent Research.
