• Medientyp: E-Artikel
  • Titel: Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons
  • Beteiligte: Cotter, Laurent; Yu, Feifan; Roqueviere, Sylvain; Duchesne de Lamotte, Juliette; Krupp, Johannes; Dong, Min; Nicoleau, Camille
  • Erschienen: Springer Science and Business Media LLC, 2023
  • Erschienen in: Communications Biology
  • Sprache: Englisch
  • DOI: 10.1038/s42003-023-04495-w
  • ISSN: 2399-3642
  • Schlagwörter: General Agricultural and Biological Sciences ; General Biochemistry, Genetics and Molecular Biology ; Medicine (miscellaneous)
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  • Beschreibung: <jats:title>Abstract</jats:title><jats:p>Botulinum neurotoxins (BoNTs) have been widely used clinically as a muscle relaxant. These toxins target motor neurons and cleave proteins essential for neurotransmitter release like Synaptosomal-associated protein of 25 kDa (SNAP-25). In vitro assays for BoNT testing using rodent cells or immortalized cell lines showed limitations in accuracy and physiological relevance. Here, we report a cell-based assay for detecting SNAP-25-cleaving BoNTs by combining human induced Pluripotent Stem Cells (hiPSC)-derived motor neurons and a luminescent detection system based on split NanoLuc luciferase. This assay is convenient, rapid, free-of-specialized antibodies, with a detection sensitivity of femtomolar concentrations of toxin, and can be used to study the different steps of BoNT intoxication.</jats:p>
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