The ECHDC1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of Raji B lymphocytes with targeted disruptions in the ECHDC1 gene. This loss-of-function model enables study of ethylmalonyl-CoA decarboxylase in a human Burkitt lymphoma background. The heterogeneous genotype reflects cellular variability in response to ECHDC1 disruption, reducing clone-specific artifacts and making it suitable for pooled functional screens and bulk metabolic assays. The knockout was engineered using CRISPR/Cas9 reagents to introduce disruptive edits into the ECHDC1 locus.
Raji cells are a human B lymphocyte line derived from Burkitt lymphoma, EBV-positive and lymphoblastoid. They exhibit mature B-cell features, including antibody production, and are widely used in immunology and cancer research. The EBV-driven immortalization allows large-scale culture and high-throughput screening. Raji cells model B-cell malignancies, with MYC translocation and altered metabolism. ECHDC1 knockout in these cells helps dissect metabolic vulnerabilities in aggressive lymphomas and explore branched-chain fatty acid metabolism in immune cells.
ECHDC1 decarboxylates ethylmalonyl-CoA to butyryl-CoA and CO2, a critical step in branched-chain fatty acid catabolism linked to propanoate metabolism and fatty acid ??-oxidation. This reaction regulates ethylmalonic acid levels and mitochondrial energy metabolism. ECHDC1 expression is transcriptionally controlled by PPAR??, PGC-1??, and SREBP1c, and is integrated with AMPK signaling. The product butyryl-CoA fuels fatty acid elongation and influences histone acylation, connecting metabolism to epigenetic regulation. ECHDC1 interacts with ECHS1 and other mitochondrial enoyl-CoA hydratases, forming a CoA-dependent complex to channel intermediates. Disrupting ECHDC1 thus perturbs ethylmalonate metabolism and may alter branched-chain amino acid degradation and fatty acid oxidation.
In Raji Burkitt lymphoma cells, ECHDC1 knockout enables investigation of mitochondrial fatty acid metabolism in cancer proliferation. These cells have high MYC-driven anabolic demand; while glycolysis is prominent, fatty acid oxidation can support tumorigenesis. ECHDC1 ablation helps determine if branched-chain fatty acid catabolism is essential for energy or biosynthesis in malignant B cells. This model also provides insights into ethylmalonic aciduria, where ethylmalonyl-CoA accumulation is pathogenic. Additionally, the EBV-positive background allows study of viral latency effects on lipid metabolism.
The cells support metabolomics profiling of ethylmalonyl-CoA and butyryl-CoA, ethylmalonyl-CoA decarboxylase activity assays, and fatty acid oxidation flux measurements. Western blotting and RT-qPCR confirm ECHDC1 disruption. They enable functional genomic screens in B cells, such as CRISPR modifier studies or drug-sensitivity assays. Researchers in metabolic disorders, cancer metabolism, or B-cell biology will find this a versatile tool. For further information and technical support, please contact Ascent Research.
