MECR Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of Raji B lymphocytes with targeted disruption of the MECR gene. This loss-of-function model enables investigation of mitochondrial trans-2-enoyl-CoA reductase, a key enzyme in mitochondrial fatty acid synthesis (mtFAS), within a Burkitt’s lymphoma background. The polyclonal format preserves cellular diversity and provides a robust tool for functional genomics and metabolic pathway analysis without clonal selection artifacts.
The parental Raji cell line is an Epstein-Barr virus (EBV)-transformed lymphoblastoid line derived from a Burkitt’s lymphoma patient. Raji cells grow in suspension and serve as a well-established model of non-Hodgkin B-cell lymphoma, maintaining active humoral immune response pathways. They are widely used to study B-cell receptor signaling, apoptosis, and oncogenesis, making them particularly relevant for exploring metabolic dependencies in lymphomagenesis.
MECR catalyzes the NADPH-dependent reduction of trans-2-enoyl-CoA to acyl-CoA, a critical step in mtFAS that generates octanoyl-ACP, the precursor for lipoic acid biosynthesis. Lipoic acid is essential for lipoylation of mitochondrial dehydrogenase complexes, including the E2 subunits of pyruvate dehydrogenase (DLAT) and ??-ketoglutarate dehydrogenase (OGDH), as well as the glycine cleavage system. MECR operates within a complex comprising MCAT, OXSM, NDUFAB1, CBR4, HTD2, LIAS, and LIPT1. Its expression is regulated by mitochondrial biogenesis signaling factors such as PPARGC1A (PGC-1??), NRF1, and TFAM, linking mtFAS to broader metabolic control.
Knockout of MECR in Raji cells disrupts mtFAS, abolishing octanoyl-ACP production and impairing lipoic acid biosynthesis. Consequently, lipoylation of DLAT and PDHX (PDH subunits) and OGDH is compromised, inactivating these key TCA cycle-linked dehydrogenases. This metabolic perturbation shifts cellular energy metabolism and may alter mitochondrial membrane potential and ATP production. As Burkitt’s lymphoma cells are highly glycolytic, this model uniquely reveals the functional importance of mitochondrial metabolism and lipoic acid-dependent regulation in B-cell malignancies.
Applications include studying mitochondrial metabolism in B-cell lymphoma, screening for modulators of mtFAS, and modeling MECR-related neurodegeneration (MEPAN syndrome) in a B-cell context. Researchers can perform Western blot analysis of MECR and lipoylated DLAT and OGDH, measure mitochondrial respiration via Seahorse assay, quantify PDH activity, assess mitochondrial membrane potential by TMRM flow cytometry, and profile acyl-CoA levels by LC-MS. This polyclonal knockout population serves as a versatile platform for metabolic research. For further technical details, please contact Ascent Research.
