High-throughput screening for a SARS-CoV-2 frameshifting inhibitor using a cell-free protein synthesis system.
Kodai MachidaRin TanakaSeraya MikiShotaro NosedaMayumi Yuasa-SunagawaHiroaki ImatakaPublished in: BioTechniques (2024)
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.
Keyphrases
- sars cov
- cell free
- respiratory syndrome coronavirus
- induced apoptosis
- circulating tumor
- cell cycle arrest
- single cell
- oxidative stress
- single molecule
- gene expression
- mesenchymal stem cells
- bone marrow
- cell proliferation
- coronavirus disease
- endoplasmic reticulum stress
- signaling pathway
- high resolution
- molecularly imprinted
- energy transfer
- solid phase extraction