Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons.
Laurent CotterFeifan YuSylvain RoqueviereJuliette Duchesne de LamotteJohannes KruppMin DongCamille NicoleauPublished in: Communications biology (2023)
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.
Keyphrases
- induced pluripotent stem cells
- high throughput
- loop mediated isothermal amplification
- spinal cord
- single cell
- induced apoptosis
- sensitive detection
- endothelial cells
- real time pcr
- skeletal muscle
- palliative care
- label free
- cell therapy
- quantum dots
- oxidative stress
- endoplasmic reticulum stress
- heat shock protein
- cell proliferation
- pluripotent stem cells