The ECHS1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from Raji B lymphocytes to enable loss-of-function studies of the mitochondrial enoyl-CoA hydratase ECHS1. This polyclonal pool, produced via CRISPR/Cas9-mediated gene disruption, provides a versatile model for investigating ECHS1 deficiency in a lymphoma background.
The Raji cell line is an immortalized B lymphocyte model derived from human Burkitt’s lymphoma, Epstein-Barr virus (EBV) positive, and widely used for immunological and cancer research. As a lymphoblastoid line, Raji retains key B cell characteristics and serves as a robust platform for studying B cell lymphoma pathogenesis, viral oncogenesis, and cellular metabolism.
ECHS1 encodes a mitochondrial short-chain enoyl-CoA hydratase that catalyzes the second step of fatty acid ??-oxidation and participates in the catabolism of branched-chain amino acids (valine, leucine, isoleucine). Transcription is regulated by the nuclear receptors and coactivators PPAR??, PGC-1??, NRF1, and ERR??. Within the matrix, ECHS1 hydrates short-chain enoyl-CoA thioesters to (S)-3-hydroxyacyl-CoA, substrates for the trifunctional protein (HADHA/HADHB) and short/branched chain acyl-CoA dehydrogenase (ACADSB), with electrons transferred via ETF. Interacting factors include the mitochondrial import receptor Tom20 and 3-hydroxyacyl-CoA dehydrogenase. Disruption of ECHS1 leads to accumulation of upstream intermediates, diminished acetyl-CoA and ATP synthesis, and reduced NADH/FADH2 generation, recapitulating the metabolic hallmarks of ECHS1 deficiency.
In Raji lymphoma cells, which exhibit high glycolytic flux typical of Burkitt’s lymphoma, ECHS1 knockout allows assessment of metabolic dependencies on mitochondrial oxidative pathways. The EBV-positive context further enables exploration of how viral oncoproteins modulate lipid catabolism, potentially revealing synthetic lethal interactions or metabolic vulnerabilities relevant to B cell malignancy therapy.
Researchers can employ this polyclonal knockout population for fatty acid oxidation defect modeling, metabolic reprogramming analysis in B cell lymphoma, mitochondrial dysfunction characterization, nutrient dependency screens, and drug sensitivity testing. Representative assays include ECHS1 immunoblotting, radiolabeled palmitate oxidation, Seahorse stress tests, ATP measurement, and metabolomic profiling, providing comprehensive tools for cancer metabolism studies and mitochondrial biology research. For further information, please contact Ascent Research.
