The ECI1 Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal population derived from the human HAP1 cell line, enabling disruption of the ECI1 gene. This polyclonal knockout model provides a heterogeneous pool of cells with targeted gene disruption, facilitating robust loss-of-function studies without clonal selection. It allows investigation of mitochondrial unsaturated fatty acid ??-oxidation in a human cellular context.
HAP1 is a near-haploid human chronic myeloid leukemia cell line derived from KBM-7, characterized by a predominantly haploid karyotype that simplifies CRISPR/Cas9-mediated knockout. The line is p53-deficient and adapted to suspension culture, facilitating efficient genome editing and scalability for functional genomics and high-throughput assays. Its metabolic profile makes it well-suited for studying mitochondrial energy pathways.
ECI1 encodes mitochondrial enoyl-CoA delta isomerase 1, an auxiliary enzyme essential for ??-oxidation of unsaturated fatty acids. It isomerizes 3-cis and 3-trans enoyl-CoA to 2-trans enoyl-CoA, a prerequisite for further processing by HADHA/HADHB and ACAA2 within the ??-oxidation spiral. ECI1 is regulated by PPARA and PPARGC1A and functions upstream of acetyl-CoA, NADH, and FADH2 generation, linking fatty acid catabolism to mitochondrial electron transport. Interactors include VLCAD and other ??-oxidation enzymes, forming a coordinated pathway with CPT1, CPT2, and HADHA/HADHB.
In HAP1 cells, ECI1 disruption models ECI1 deficiency, a metabolic disorder associated with Reye-like syndrome, rhabdomyolysis, and metabolic acidosis. The haploid background ensures uniform knockout for studying impaired unsaturated fatty acid utilization and its consequences on mitochondrial energetics. The p53 deficiency and suspension growth enable large-scale metabolic flux analyses and compound screening, making this model ideal for investigating the pathophysiology of fatty acid oxidation disorders.
This ECI1 knockout pool supports applications in unsaturated fatty acid metabolism research, mitochondrial ??-oxidation disorder studies, and metabolic disease drug development. Assays include RT-qPCR and Western blot for knockdown confirmation, fatty acid oxidation measurements, enzyme activity assays, metabolic flux analysis, and immunofluorescence. Researchers can use this model to dissect ECI1??s role in energy production and screen therapeutic interventions. For further information, contact Ascent Research.
