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

Commun Biol. 2023 Jan 30;6(1):122. doi: 10.1038/s42003-023-04495-w.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Transport
  • Botulinum Toxins, Type A* / metabolism
  • Botulinum Toxins, Type A* / toxicity
  • Humans
  • Induced Pluripotent Stem Cells* / metabolism
  • Motor Neurons / metabolism

Substances

  • Botulinum Toxins, Type A