Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons

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 immortali...

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Autores principales: Cotter, Laurent, Yu, Feifan, Roqueviere, Sylvain, Duchesne de Lamotte, Juliette, Krupp, Johannes, Dong, Min, Nicoleau, Camille
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9886929/
https://www.ncbi.nlm.nih.gov/pubmed/36717690
http://dx.doi.org/10.1038/s42003-023-04495-w
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author Cotter, Laurent
Yu, Feifan
Roqueviere, Sylvain
Duchesne de Lamotte, Juliette
Krupp, Johannes
Dong, Min
Nicoleau, Camille
author_facet Cotter, Laurent
Yu, Feifan
Roqueviere, Sylvain
Duchesne de Lamotte, Juliette
Krupp, Johannes
Dong, Min
Nicoleau, Camille
author_sort Cotter, Laurent
collection PubMed
description 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.
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spelling pubmed-98869292023-02-01 Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons Cotter, Laurent Yu, Feifan Roqueviere, Sylvain Duchesne de Lamotte, Juliette Krupp, Johannes Dong, Min Nicoleau, Camille Commun Biol Article 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. Nature Publishing Group UK 2023-01-30 /pmc/articles/PMC9886929/ /pubmed/36717690 http://dx.doi.org/10.1038/s42003-023-04495-w Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Cotter, Laurent
Yu, Feifan
Roqueviere, Sylvain
Duchesne de Lamotte, Juliette
Krupp, Johannes
Dong, Min
Nicoleau, Camille
Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons
title Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons
title_full Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons
title_fullStr Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons
title_full_unstemmed Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons
title_short Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons
title_sort split luciferase-based assay to detect botulinum neurotoxins using hipsc-derived motor neurons
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9886929/
https://www.ncbi.nlm.nih.gov/pubmed/36717690
http://dx.doi.org/10.1038/s42003-023-04495-w
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