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These toxins target motor neurons and cleave proteins essential for neurotransmitter release like Synaptosomal-associated protein of 25\u2009kDa (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>","DOI":"10.1038\/s42003-023-04495-w","type":"journal-article","created":{"date-parts":[[2023,1,30]],"date-time":"2023-01-30T07:03:45Z","timestamp":1675062225000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons"],"prefix":"10.1038","volume":"6","author":[{"given":"Laurent","family":"Cotter","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feifan","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sylvain","family":"Roqueviere","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juliette","family":"Duchesne de Lamotte","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johannes","family":"Krupp","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1744-7293","authenticated-orcid":false,"given":"Min","family":"Dong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9803-1053","authenticated-orcid":false,"given":"Camille","family":"Nicoleau","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2023,1,30]]},"reference":[{"key":"4495_CR1","doi-asserted-by":"publisher","first-page":"535","DOI":"10.1038\/nrmicro3295","volume":"12","author":"O Rossetto","year":"2014","unstructured":"Rossetto, O., Pirazzini, M. & Montecucco, C. 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Harvard Medical School has filed a patent application covering the split NanoLuc-based assays for detecting botulinum neurotoxins, with F.Y. and M.D. as co-inventors.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"The methods were performed in accordance with relevant guidelines and regulations and approved by The National Agency for the Safety of Medicines and Health Products (ANSM).","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical approval"}}],"article-number":"122"}}