MCCC2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Raji B lymphocyte cell line, designed for targeted disruption of the MCCC2 gene. This polyclonal pool provides a heterogeneous loss-of-function model, avoiding the limitations of single-cell clones. The gene-edited cells enable robust investigation of MCCC2??s role in leucine degradation and mitochondrial metabolism within an immune context.
The Raji cell line is a human Burkitt lymphoma B lymphocyte model, Epstein-Barr virus (EBV) positive, widely used in immunology and cancer research. Raji cells exhibit rapid suspension growth, antibody production, and antigen-presentation capacity, offering a physiologically relevant background for studying metabolic regulation in B cells. The EBV-driven transformation phenotype and well-characterized genetics facilitate dissection of how metabolic disruptions intersect with immune function.
MCCC2 encodes the beta subunit of 3-methylcrotonyl-CoA carboxylase (MCCC), which partners with MCCC1 and biotin to carboxylate 3-methylcrotonyl-CoA to 3-methylglutaconyl-CoA in leucine degradation. The MCCC holoenzyme depends on holocarboxylase synthetase (HLCS) for biotin attachment, and activity is modulated by leucine concentration and mTORC1 signaling. PPARGC1A transcriptionally upregulates MCCC2, while downstream metabolites include 3-methylglutaconyl-CoA, HMG-CoA, and acetoacetate. Operating alongside branched-chain amino acid transaminase and HMG-CoA lyase, MCCC2 integrates leucine catabolism with mitochondrial bioenergetics.
MCCC2 disruption in Raji cells abolishes 3-methylcrotonyl-CoA carboxylase activity, causing accumulation of 3-methylcrotonyl-CoA and 3-hydroxyisovaleric acid??hallmarks of 3-methylcrotonyl-CoA carboxylase deficiency. This metabolic blockade impairs leucine-dependent energy production, likely affecting mitochondrial respiration and B cell proliferation. The polyclonal knockout pool recreates key disease-relevant metabolic perturbations, facilitating research into metabolic acidosis, hypoglycemia, and neurodevelopmental defects. By eliminating MCCC function, these cells enable studies of compensatory metabolic pathways and the impact of leucine deprivation on immune homeostasis.
Researchers can employ MCCC2 Knockout Raji Polyclonal Cells for metabolic flux analysis via LC-MS metabolomics, enzymatic activity assays, and Seahorse mitochondrial stress testing. The model supports drug screening for metabolic disorders, mTORC1 nutrient-sensing studies, and mitochondrial dysfunction research. Typical readouts include Western blotting, RT-qPCR, and leucine tolerance assays. For additional information or to order, please contact Ascent Research.
