The ECHS1 Knockout K-562 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of K-562 cells that harbor a targeted disruption of the ECHS1 gene. This loss-of-function model is generated by introducing sequence-specific guide RNAs and Cas9 nuclease to ablate ECHS1 expression, providing a heterogeneous pool of edited cells suitable for functional studies of mitochondrial fatty acid ??-oxidation and branched-chain amino acid catabolism in a leukemic background.
The host cell line, K-562, is a human chronic myelogenous leukemia (CML) cell line originally derived from a female patient in blast crisis. K-562 cells display characteristics of both erythroid and myeloid lineages and carry the BCR-ABL1 fusion oncogene, making them a widely employed model for investigating hematopoietic differentiation, CML pathogenesis, and therapeutic responses. The cell line??s robust growth and well-characterized biology facilitate reproducible experiments in mitochondrial metabolism research.
ECHS1 encodes the mitochondrial short-chain enoyl-CoA hydratase that catalyzes the hydration of trans-2-enoyl-CoA to L-3-hydroxyacyl-CoA, a key step in the fatty acid ??-oxidation spiral and branched-chain amino acid catabolism. This reaction is essential for the generation of acetyl-CoA and TCA cycle intermediates. ECHS1 expression is regulated by PPARA, PPARGC1A, and ESRRA, and its protein product interacts with the mitochondrial trifunctional protein subunits HADHA and HADHB, as well as HSD17B10. Disruption of ECHS1 blocks ??-oxidation, leading to accumulation of enoyl-CoA substrates and impaired mitochondrial energy production, phenocopying the metabolic deficiency seen in short-chain enoyl-CoA hydratase deficiency and Leigh syndrome.
In K-562 leukemia cells, ECHS1 knockout creates a valuable model to dissect the role of mitochondrial fatty acid oxidation in CML biology, metabolic reprogramming, and drug sensitivity. The BCR-ABL1-driven oncogenic signaling is known to alter cellular metabolism, and loss of ECHS1 may exacerbate mitochondrial dysfunction, influence redox balance, and modulate apoptotic responses. This model is particularly relevant for studying the interplay between oncogenic kinase signaling and mitochondrial energy metabolism, as well as for investigating potential therapeutic vulnerabilities in leukemias with altered lipid utilization.
Researchers can employ this polyclonal knockout population in diverse experimental workflows, including immunoblotting to confirm ECHS1 depletion, RT-qPCR for transcriptional analysis, Seahorse metabolic flux assays to measure oxidative phosphorylation and glycolysis, LC-MS-based acyl-CoA profiling to assess substrate accumulation, and cell viability or apoptosis assays under nutrient stress or drug treatment. These cells are suitable for screening metabolic inhibitors, analyzing branched-chain amino acid metabolic flux, and evaluating the mitochondrial basis of drug resistance. For further information, please contact Ascent Research.
