High-throughput screening for a SARS-CoV-2 frameshifting inhibitor using a cell-free protein synthesis system

Biotechniques. 2024 Apr;76(4):161-168. doi: 10.2144/btn-2023-0102. Epub 2024 Jan 31.

Abstract

Programmed-1 ribosomal frameshifting (-1 PRF) is a translational mechanism adopted by some viruses, including SARS-CoV-2. To find a compound that can inhibit -1 PRF in SARS-CoV-2, we set up a high-throughput screening system using a HeLa cell extract-derived cell-free protein synthesis (CFPS) system. A total of 32,000 compounds were individually incubated with the CFPS system programmed with a -1 PRF-EGFP template. Several compounds were observed to decrease the -1 PRF-driven fluorescence, and one of them had some suppressive effect on -1 PRF of a SARS-CoV-2 genome sequence in transfected cells. Thus the CFPS system can be used as a tool for a high-throughput screening of chemicals.

Keywords: SARS-CoV-2; cell-free protein synthesis; drug discovery; programmed-1 ribosomal frameshifting; screening.

MeSH terms

  • COVID-19*
  • Frameshifting, Ribosomal
  • HeLa Cells
  • High-Throughput Screening Assays
  • Humans
  • Protein Biosynthesis
  • SARS-CoV-2*