The ECI2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population engineered to disrupt the ECI2 gene in the near-haploid HAP1 human cell line. This loss-of-function model enables detailed investigation of mitochondrial enoyl-CoA delta isomerase 2, a pivotal enzyme in the ??-oxidation of unsaturated fatty acids. The polyclonal format ensures a diverse repertoire of knockout alleles, providing robust and reproducible functional data ideal for high-throughput screening applications without the need for single-cell cloning.
HAP1 is a near-haploid cell line derived from the KBM-7 chronic myeloid leukemia (CML) line, originally isolated from a male patient in blast crisis. Apart from chromosome 8, these cells retain a haploid karyotype, which simplifies genetic manipulation and phenotypic interpretation by eliminating confounding diploid gene dosage effects. This attribute has established HAP1 as a powerful model for functional genomics and CRISPR-based screens. Its CML origin further makes it a physiologically relevant system for studying metabolic pathways in leukemia, including fatty acid oxidation.
The ECI2 gene encodes mitochondrial enoyl-CoA delta isomerase 2, a critical enzyme that catalyzes the isomerization of 3-cis/trans-enoyl-CoA intermediates to 2-trans-enoyl-CoA, an essential step in the mitochondrial ??-oxidation of unsaturated fatty acids. ECI2 activity is regulated by the transcription factors PPARA, PPARG, and the coactivator PPARGC1A, linking its expression to peroxisome proliferator-activated receptor signaling. Downstream, ECI2 contributes to the generation of acetyl-CoA, ATP, and acylcarnitine species. In the broader pathway, it functions alongside ACADVL, ECHS1, HADH, and ACAA2. Its disruption blocks isomerization, causing accumulation of upstream intermediates and reducing lipid-derived energy production.
In the HAP1 context, where cellular energy balance heavily relies on fatty acid oxidation, loss of ECI2 severely impairs the catabolism of unsaturated fats. This leads to altered acylcarnitine profiles, diminished mitochondrial respiration, and decreased ATP levels, potentially compromising cell proliferation under lipid-rich conditions. The near-haploid background of HAP1 cells allows clear genotype-phenotype correlations, making this polyclonal knockout population an ideal tool to dissect the metabolic vulnerabilities of CML cells. It also facilitates the study of compensatory metabolic shifts that may occur upon disruption of unsaturated fatty acid oxidation.
Researchers can apply this knockout model to metabolic profiling of lipid disorders, cancer metabolism studies, and drug screening for ??-oxidation modulators. Representative assays include acylcarnitine LC-MS analysis, fatty acid oxidation flux measurements, Seahorse respirometry, ATP quantification, and lipid-dependent proliferation assays. The polyclonal nature of the cell population supports high-throughput experimental designs and ensures robust statistical power across replicates. This product is particularly suited for investigating metabolic reprogramming in leukemia and screening compounds that target mitochondrial fatty acid oxidation. For further information, please contact Ascent Research.
