Split Luciferase-based Assay to Detect Botulinum Neurotoxins Using HiPSC-derived Motor Neurons

Overview

Journal Commun Biol

Specialty Biology

Date 2023 Jan 30

PMID 36717690

Authors

Laurent Cotter

Feifan Yu

Sylvain Roqueviere

Juliette Duchesne de Lamotte

Johannes Krupp

Min Dong

Camille Nicoleau

Affiliations

Soon will be listed here.

Abstract

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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