The ECI1 Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphocyte line, carrying a targeted disruption of the ECI1 gene. This product provides a renewable pool of gene-edited cells for loss-of-function studies, enabling robust investigation of enoyl-CoA delta isomerase 1 function without clonal selection bias. The polyclonal nature preserves population diversity, making it suitable for experiments requiring bulk cellular responses to ECI1 ablation. CRISPR/Cas9-mediated gene disruption introduces permanent modifications at the ECI1 locus, allowing stable ablation of the encoded enzyme involved in mitochondrial unsaturated fatty acid catabolism.
The parental Raji cell line originates from a Burkitt lymphoma patient and maintains Epstein-Barr virus positivity, growing in suspension with lymphoblastoid morphology. As a human B lymphocyte model, Raji cells are integral to studying antibody production, antigen presentation, and immune surveillance mechanisms. Their malignant origin and ease of culture make them widely used in cancer biology and immunology research. The lymphoblast context offers a relevant system for examining how metabolic pathways, particularly fatty acid oxidation, intersect with B-cell proliferation and survival, especially under the influence of viral oncogenesis.
ECI1 encodes enoyl-CoA delta isomerase 1, a mitochondrial enzyme essential for the beta-oxidation of unsaturated fatty acids. It catalyzes the isomerization of 3-cis and 3-trans enoyl-CoA esters to 2-trans enoyl-CoA, a prerequisite for continued chain shortening by acyl-CoA dehydrogenases. This protein functions in complex with very long-chain acyl-CoA dehydrogenase (VLCAD), medium-chain acyl-CoA dehydrogenase (MCAD), and the mitochondrial trifunctional protein (MTP), while receiving electrons from electron transfer flavoprotein (ETF). ECI1 expression is transcriptionally regulated by PPAR??, PPAR??, and PGC-1??, and its activity is influenced by energy sensors AMPK and SIRT1. Downstream, ECI1-dependent flux generates acetyl-CoA, NADH, FADH2, and medium-chain acyl-CoAs, fueling ATP production and anabolic processes. Disruption of ECI1 thus halts unsaturated fatty acyl-CoA processing, leading to accumulation of 3-cis/trans intermediates and blunted lipid-derived energy output.
In the Raji Burkitt lymphoma model, ECI1 knockout creates a unique system to probe the reliance of malignant B cells on fatty acid beta-oxidation. Lymphoma cells often exhibit metabolic reprogramming, and reliance on lipid catabolism can support rapid proliferation and survival under stress. Loss of ECI1 may compromise mitochondrial respiration fueled by unsaturated fatty acids, potentially sensitizing cells to metabolic stress or altering oncogenic signaling. This model allows dissection of whether ECI1-mediated isomerization is a metabolic liability in EBV-driven lymphomagenesis, offering insights into how lipid metabolism supports lymphoblastoid cell maintenance. The interplay between ECI1 and upstream regulators like AMPK and SIRT1 may further reveal how nutrient sensing intersects with fatty acid utilization in cancer.
Researchers can leverage this polyclonal knockout product for detailed functional analyses, including Western blotting to confirm ECI1 protein loss, RT-qPCR for mRNA assessment, and fatty acid oxidation flux assays using radiolabeled or stable isotope-labeled palmitate or oleate. Mitochondrial respiration defects can be quantified via Seahorse analysis, while lipidomic profiling reveals accumulation of unsaturated acyl-CoA intermediates. Cell proliferation, apoptosis, and drug sensitivity studies with metabolic inhibitors (e.g., etomoxir, perhexiline) can identify synthetic lethal interactions. This model is also suitable for screening candidate molecules targeting fatty acid metabolism in lymphoma. For additional technical information or support, please contact Ascent Research.
