The ECI2 Knockout K-562 Polyclonal Cells consist of a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population in which the ECI2 (enoyl-CoA delta isomerase 2) locus has been targeted for loss-of-function studies. This polyclonal knockout model is generated from the human K-562 suspension lymphoblastoid cell line and is designed for investigating the role of mitochondrial unsaturated fatty acid oxidation in leukemia cell biology. The heterogeneous population allows for bulk analysis of ECI2 disruption effects, avoiding clonal artifacts and providing a representation of the knockout impact across a diverse genetic background.
The parental K-562 cell line is a widely studied human chronic myelogenous leukemia (CML) model originally isolated from the pleural effusion of a 53-year-old female in blast crisis. K-562 cells are non-adherent, grow in suspension, and display a lymphoblastoid morphology. They express markers of multiple hematopoietic lineages, reflecting their origin from a multipotent progenitor. This cell line is extensively used to study CML pathophysiology, signal transduction, and drug resistance mechanisms, providing a robust platform for functional genomics studies in the blast crisis phase of CML.
ECI2 (enoyl-CoA delta isomerase 2) catalyzes the isomerization of 3-cis and 2-trans-enoyl-CoA esters to 2-trans-enoyl-CoA, a necessary step in mitochondrial beta-oxidation of unsaturated fatty acids. This reaction follows fatty acid activation by acyl-CoA synthetases and carnitine-dependent transport via CPT1/2 and carnitine-acylcarnitine translocase. Within the beta-oxidation complex, ECI2 works alongside very long-chain acyl-CoA dehydrogenase (VLCAD), enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and the trifunctional protein complex. Its transcription is regulated by PPARalpha (PPARA)/RXR heterodimers and PGC-1alpha, responding to PPARalpha agonists such as fenofibrate. The isomerization product continues through the beta-oxidation spiral to yield acetyl-CoA for the TCA cycle and ATP synthesis. Disruption of ECI2 leads to inefficient unsaturated fatty acid oxidation, accumulation of aberrant enoyl-CoA intermediates, and compromised cellular energy balance.
In the K-562 leukemia background, ECI2 knockout provides a defined loss-of-function model to interrogate the dependency of blast crisis CML cells on fatty acid oxidation. Leukemia cells often exhibit metabolic reprogramming, and the reliance on mitochondrial beta-oxidation for energy and biosynthetic precursors can represent a therapeutic vulnerability. The polyclonal nature of this knockout population preserves some heterogeneity, making it suitable for studying the overall impact of ECI2 deletion on cellular fitness, proliferation, and survival under metabolic stress. By impairing unsaturated fatty acid catabolism, this model can reveal compensatory metabolic shifts and potential synthetic lethal interactions.
Researchers can employ these polyclonal ECI2 knockout K-562 cells in a wide array of functional assays to dissect the metabolic underpinnings of leukemia. Typical applications include measurements of fatty acid oxidation rates using 14C-oleate tracing, assessment of mitochondrial respiration via Seahorse metabolic flux analysis (oxygen consumption rate, OCR), ATP quantification assays, and cell proliferation analyses. Target gene disruption can be confirmed by western blotting or RT-qPCR. Additionally, lipidomic and metabolomic profiling can uncover accumulated intermediates and altered metabolic networks. This model is particularly suited for drug target validation in metabolic disorders and for evaluating the efficacy of PPARalpha agonists or other modulators. For further details and technical support, please contact Ascent Research.
