ECH1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human embryonic kidney cell line HEK293T. This product provides a loss-of-function model for studying the peroxisomal enzyme enoyl-CoA hydratase (ECH1). The polyclonal format ensures a heterogeneous mix of gene-disrupted cells, enabling robust assessment of ECH1-dependent phenotypes without clonal selection biases.
Host cell background: HEK293T cells are a widely used derivative of the HEK293 line, stably expressing the SV40 large T antigen. These adherent epithelial-like cells exhibit high transfectability and are commonly employed for viral production, protein expression, and functional genomics studies. Their rapid growth and ease of manipulation make them an ideal chassis for CRISPR-based knockout studies, particularly when investigating metabolic pathways.
Molecular function: ECH1 catalyzes the second step of peroxisomal fatty acid ??-oxidation, hydrating trans-2-enoyl-CoA to L-3-hydroxyacyl-CoA. This reaction is part of the peroxisomal ??-oxidation cycle, which degrades very long-chain, branched-chain, and bile acid intermediates. ECH1 activity is regulated by nuclear receptors such as PPAR??, FXR, and LXR, and is influenced by fasting and high-fat diet. It functions downstream of peroxisomal acyl-CoA oxidases and interacts with the multifunctional protein (MFP), sterol carrier protein x (SCPx), and D-bifunctional protein (DBP) to channel metabolites toward acetyl-CoA production.
Model significance: In the HEK293T context, ECH1 knockout allows dissection of peroxisomal lipid metabolism in a genetically tractable system. Since HEK293T cells lack robust peroxisomal ??-oxidation compared to liver cells, the knockout phenotype may sensitize cells to lipid stress, enabling assays that measure viability under excess fatty acid conditions. This model is valuable for studying the molecular pathogenesis of peroxisomal disorders, including metabolic encephalopathies and fatty acid oxidation defects, where ECH1 dysfunction is implicated.
Research applications: Researchers can employ these polyclonal knockout cells in a variety of experiments: western blotting and RT-qPCR to confirm ECH1 loss; peroxisomal ??-oxidation activity assays using radiolabeled or fluorescent substrates; immunofluorescence co-localization with peroxisomal markers; lipidomics and metabolic flux analysis to profile altered lipid species; and viability assays under lipid overload to assess metabolic compensation. The polyclonal nature also provides a population-level view of gene disruption effects, suitable for screening or pooled analyses. For further information or customized support, please contact Ascent Research.
