The ECHS1 Knockout HEK293T Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population in the HEK293T human embryonic kidney background. This heterogeneous loss-of-function model enables investigation of the mitochondrial enzyme ECHS1 (enoyl-CoA hydratase, short chain 1). CRISPR/Cas9-mediated gene disruption yields a population-based knockout tool suitable for diverse metabolic and mitochondrial studies without clonal selection.
HEK293T cells are SV40 large T-antigen-transformed human embryonic kidney epithelial cells, valued for high transfection efficiency and robust protein expression. Their adherent epithelial morphology and active mitochondrial metabolism make them a suitable platform for examining fatty acid oxidation and mitochondrial dysfunction. In the setting of ECHS1 knockout, this host line permits biochemical and metabolic flux analyses to dissect the enzyme’s role in energy homeostasis.
ECHS1 encodes a mitochondrial matrix enzyme that catalyzes the second step of fatty acid ??-oxidation, converting trans-2-enoyl-CoA to 3-hydroxyacyl-CoA. It is essential for short-chain fatty acid and branched-chain amino acid (valine, leucine, isoleucine) degradation. Transcriptionally regulated by PPARA, PGC-1??, and AMPK signaling, ECHS1 activity drives acetyl-CoA and NADH production to fuel the TCA cycle and oxidative phosphorylation. It physically interacts with the mitochondrial trifunctional protein (HADHA/HADHB) and coenzyme A, collaborating with ACADM, ACADS, and ACAT1 within the ??-oxidation pathway.
Loss of ECHS1 function impairs the breakdown of short-chain fatty acids and branched-chain amino acids, causing accumulation of toxic acyl-CoA species and reduced ATP synthesis. This metabolic disruption models features of human mitochondrial disorders such as Leigh syndrome and paroxysmal exercise-induced dystonia. In HEK293T cells, ECHS1 knockout compromises oxidative phosphorylation and sensitizes cells to metabolic stress, providing a relevant system for mechanistic studies and therapeutic screening.
This polyclonal knockout population supports applications including the study of mitochondrial fatty acid oxidation disorders, metabolic pathway analysis, and compound screening for Leigh syndrome. Key assays include Western blotting, RT-qPCR, immunofluorescence, Seahorse metabolic flux analysis, fatty acid oxidation assays, and metabolomic profiling of acyl-CoA intermediates. Enzyme activity and cell viability under metabolic stress further characterize the knockout phenotype. For additional information or custom inquiries, please contact Ascent Research.
