ECHDC3 Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the K-562 human chronic myelogenous leukemia cell line. These cells feature targeted disruption of the ECHDC3 gene, which encodes a mitochondrial enoyl-CoA hydratase/isomerase that catalyzes hydration of trans-enoyl-CoA intermediates in fatty acid beta-oxidation. The polyclonal format offers a heterogeneous pool of gene edits, enabling functional studies without clone-specific biases.
The K-562 host cell line was established from the bone marrow of a 53-year-old female with chronic myelogenous leukemia in blast crisis and is positive for the Philadelphia chromosome (BCR-ABL1 fusion). As an undifferentiated blast cell model, K-562 is widely utilized to investigate hematopoietic malignancy biology, including oncogenic signaling, apoptosis, and metabolic reprogramming. The cell line displays active glycolysis and mitochondrial respiration, making it an ideal platform to assess the impact of mitochondrial enzyme disruptions on cancer cell metabolism.
ECHDC3 functions as an enoyl-CoA hydratase/isomerase in the mitochondrial fatty acid beta-oxidation pathway, converting trans-enoyl-CoA species to 3-hydroxyacyl-CoA. Its expression is transcriptionally regulated by PPAR-alpha, PPAR-gamma, and the coactivator PGC-1alpha, master regulators of lipid metabolism. The enzyme forms functional complexes with the mitochondrial trifunctional protein subunits HADHA and HADHB and interacts with members of the acyl-CoA dehydrogenase family. Downstream, the 3-hydroxyacyl-CoA product is processed further by the intrinsic 3-hydroxyacyl-CoA dehydrogenase activity of HADHA and subsequently by beta-ketothiolase. Disruption of ECHDC3 thus impedes the orderly degradation of long-chain fatty acids, leading to reduced fatty acid oxidation capacity, potential accumulation of enoyl-CoA intermediates, and compromised mitochondrial ATP generation.
In the context of K-562 leukemia cells, ECHDC3 knockout provides a powerful tool to study the contribution of fatty acid beta-oxidation to leukemic cell survival and metabolic adaptability. Leukemia cells often rely on fatty acid oxidation to support bioenergetics and mitigate oxidative stress, especially under conditions of nutrient limitation or therapeutic challenge. The loss of ECHDC3 in a Philadelphia chromosome-positive background may reveal dependencies on mitochondrial lipid catabolism that can be exploited therapeutically, sensitizing cells to metabolic inhibitors or oxidative stress.
These polyclonal knockout cells are suitable for a broad array of metabolic assays, including measurement of fatty acid oxidation rates using radiolabeled substrates, Seahorse respirometry to quantify mitochondrial oxygen consumption, lipidomic profiling to detect pathway intermediate shifts, and ATP production assays. They enable investigations into metabolic reprogramming in leukemia, mitochondrial dysfunction related to spinocerebellar ataxia, and the evaluation of PPAR agonists or fatty acid oxidation inhibitors. Applications also extend to cell viability screening under metabolic stress conditions such as glucose deprivation or complex I inhibition. For product inquiries and technical support, please contact Ascent Research.
