The ECH1 Knockout MES-OV Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the mouse embryonic stem cell line MES-OV, with targeted disruption of the ECH1 gene. This mixed population of edited cells provides a loss-of-function model for studying ECH1-dependent processes in a pluripotent context.
The MES-OV cell line originates from the inner cell mass of 129/Sv strain blastocysts and exhibits robust self-renewal and pluripotency, with the capacity to differentiate into all three germ layers. These characteristics make it a versatile system for investigating developmental biology and cellular metabolism.
ECH1 encodes enoyl-CoA hydratase 1, a peroxisomal enzyme that catalyzes the second step of fatty acid ??-oxidation, hydrating trans-2-enoyl-CoA to 3-hydroxyacyl-CoA, while also possessing 3-hydroxyacyl-CoA dehydrogenase activity. This bifunctional enzyme is transcriptionally regulated by PPARA and peroxisomal import depends on the PEX5 receptor, interacting with PEX7. Within the peroxisomal ??-oxidation pathway, ECH1 acts downstream of ACOX1 and cooperates with HSD17B4, SCPx, and ACAA1 to degrade very long-chain fatty acids and process bile acid intermediates, generating acetyl-CoA and chain-shortened acyl-CoAs. Disruption of ECH1 blocks this catabolic sequence, causing accumulation of very long-chain fatty acids and metabolic stress.
In MES-OV pluripotent stem cells, ECH1 knockout enables dissection of peroxisomal lipid metabolism in stem cell maintenance and differentiation. Pluripotent cells depend on metabolic plasticity, and impaired fatty acid oxidation can alter cell fate decisions and redox homeostasis. This model thus provides a relevant system to study how peroxisomal defects contribute to metabolic disorders and developmental abnormalities.
This polyclonal knockout product supports diverse applications, including investigation of peroxisomal ??-oxidation mechanisms, modeling of metabolic diseases, and therapeutic testing for peroxisomal disorders. Compatible assays include Western blot and RT-qPCR for knockout validation, immunofluorescence for peroxisomal localization, GC-MS-based very long-chain fatty acid quantification, metabolic flux analysis with isotope tracers, radiolabeled palmitate oxidation assays, Seahorse respirometry, and RNA-seq for transcriptome-wide analysis. For further details or custom inquiries, please contact Ascent Research.
