Confirmation of the mode of action of an antibacterial inhibitor using regulated antisense RNA

Summary We previously demonstrated that regulated antisense RNA technology enables us to validate and identify the mode of action for some antibiotics. In this study, we have expanded the application of the regulated antisense approach to track the mode of action for a novel inhibitor of polypeptide deformylase (Pdf), which is an attractive target for the development of novel classes of antibacterial agents. We created a pdf antisense isogenic strain in Staphylococcus aureus using a TetR-regulated expression system. We demonstrated that the partial inhibition of pdf expression significantly increased the susceptibility of S. aureus to Pdf-specific inhibitor. This result provides further evidence that the TetR-regulated antisense technology is a robust tool for tracking the mode of action of novel antibacterial agents.