The ECHDC1 Knockout HAP1 Polyclonal Cells are a polyclonal population of human near-haploid HAP1 cells engineered with CRISPR/Cas9 to disrupt the ECHDC1 gene. This product provides a heterogeneous loss-of-function model, avoiding clonal isolation, which enables robust functional studies of mitochondrial fatty acid ??-oxidation. The polyclonal format ensures representation of diverse editing events across the cell pool, facilitating reproducible investigation of ECHDC1-dependent phenotypes.
The host HAP1 cell line, derived from the KBM-7 chronic myeloid leukemia line, exhibits a near-haploid karyotype that simplifies genetic knockout and functional genomics. Its fibroblast-like morphology and stable adherent growth make it ideal for high-throughput screening, metabolic assays, and advanced imaging. The near-haploid genome reduces genetic redundancy, allowing clear dissection of gene function in a mammalian context.
ECHDC1 encodes a mitochondrial enoyl-CoA hydratase that catalyzes the second step of fatty acid ??-oxidation, hydrating trans-2-enoyl-CoA to L-3-hydroxyacyl-CoA. Its expression is activated by PPAR?? and PPAR?? in response to elevated fatty acid levels, and is further modulated by insulin and glucagon. ECHDC1 functions within a multi-enzyme complex that includes acyl-CoA dehydrogenase, 3-hydroxyacyl-CoA dehydrogenase, and 3-ketoacyl-CoA thiolase, and interacts with the electron transfer flavoprotein. The reaction produces downstream metabolites acetyl-CoA, NADH, and FADH2, which feed into the TCA cycle and oxidative phosphorylation for ATP generation. Disruption of ECHDC1 arrests the ??-oxidation cycle, leading to accumulation of enoyl-CoA intermediates and impaired mitochondrial respiration.
In the HAP1 cellular background, knockout of ECHDC1 creates a powerful system to study metabolic dysregulation, as these cells depend heavily on oxidative energy metabolism. The loss of mitochondrial ??-oxidation can be directly linked to changes in mitochondrial membrane potential, lipid droplet accumulation, and ATP depletion, providing a clean phenotype for mechanistic studies. This model is particularly relevant for investigating fatty acid oxidation disorders and mitochondrial diseases, and for screening compounds that may bypass or correct the metabolic block.
Typical applications include measuring fatty acid oxidation rates using substrate-specific assays, metabolic flux analysis with Seahorse technology, ATP quantification, and mitochondrial membrane potential assessment with fluorescent probes. These cells can also be used to confirm ECHDC1 disruption via western blotting and RT-qPCR, and to explore interactive roles of PPAR-mediated signaling. The model supports drug discovery efforts targeting mitochondrial metabolism. For additional information, please contact Ascent Research.
