High-Throughput Chemical Screening for Inhibitors of Salmonella Pathogenicity Island 2

Summary Anti-virulence therapies are under active investigation as antibiotic alternatives; however, their identification from large-scale chemical libraries poses a unique challenge. The dispensability of virulence factors for growth in vitro precludes conventional, optical density-based screening methods. Here, we provide a protocol for high-throughput screening with a cell-based, promoter reporter platform. We describe the use of this method for the identification of anti-SPI-2 inhibitors specific to Salmonella Typhimurium, which may be modified to investigate other virulence factors. For complete details on the use and execution of this protocol, please refer to Tsai et al. (2020).
Graphical Abstract
Highlights • An optimized protocol to identify inhibitors of Salmonella pathogenicity island 2 • Considerations for target selection and high-throughput screen design • Guidelines for statistical analysis of chemical screening data
Anti-virulence therapies are under active investigation as antibiotic alternatives; however, their identification from large-scale chemical libraries poses a unique challenge. The dispensability of virulence factors for growth in vitro precludes conventional, optical density-based screening methods. Here, we provide a protocol for high-throughput screening with a cell-based, promoter reporter platform. We describe the use of this method for the identification of anti-SPI-2 inhibitors specific to Salmonella Typhimurium, which may be modified to investigate other virulence factors.