Identifying RNA Helicase Inhibitors Using Duplex Unwinding Assays

Methods Mol Biol. 2021:2209:53-72. doi: 10.1007/978-1-0716-0935-4_4.


RNA helicases are responsible for virtually all of RNA metabolism. Viral and bacterial pathogens typically encode their own RNA helicases. Hence, this family of enzymes is increasingly recognized as potential targets for treatment of a variety of diseases. However, the conserved structural similarities among helicase families present an obstacle to the idea of developing specific inhibitors. In order to identify potential modulators of RNA helicase activity, rapid screening approaches are needed. This has been accomplished by optimizing and adapting standard helicase assays to function in high-throughput modalities. These optimized assays have enabled the application of rapid screening approaches to be applied toward discovering helicase inhibitors. This chapter provides detailed protocols for utilizing a medium to high-throughput approach for inhibitor discovery.

Keywords: Helicase; Inhibitors; NS3; Nucleic acids.

MeSH terms

  • Enzyme Assays / methods*
  • Enzyme Inhibitors / analysis*
  • Fluorescence
  • Humans
  • RNA / chemistry*
  • RNA Helicases / antagonists & inhibitors*


  • Enzyme Inhibitors
  • RNA
  • RNA Helicases