New agents targeting multidrug resistant Staphylococcus aureus (MRSA) are urgently needed. Here, Chen et al. screened a collection of commercially available drugs for their ability to inhibit biosynthesis of staphyloxanthin, an important virulence factor that protects S. aureus from host oxidant killing. They found that the US Food and Drug Administration (FDA)-approved antifungal agent naftifine is a reversible competitive inhibitor of diapophytoene desaturase (CrtN), an enzyme that is essential for staphyloxanthin biosynthesis. Naftifine attenuated the virulence of MRSA strains in mouse models of infection, revealing CrtN as an attractive and druggable target against S. aureus infections.