Pseudomonas aeruginosais associated with the establishment of chronic, antibiotic recalcitrant infections and is considered to be a pathogen of priority. By understanding the genetic and physiological factors that drive biofilm formation, we have discovered that biofilms can be induced to disperse through the addition of nitric oxide (NO), which acts as a non-toxic signal. NO can disperse biofilms of many different bacteria and is effective against multispecies biofilms. We have shown that NO acts through modulation of the secondary messenger, c-di-GMP to control biofilm formation and that cells respond by producing the NO scavenger Fhp to limit dispersal. Furthermore, high iron levels can antagonise the activity of NO. Dispersed cells become sensitive to antibiotic treatment as well as other biocides, suggesting that NO can act in concert with other bioactive compounds to help eliminate this pathogen. We have shown that NO treatment of CF patients in the clinic can reduce biofilms of P. aeruginosa in the sputum of CF patients. To enhance the drug-like features, we have developed NO prodrugs that release NO specifically in the presence of biofilms. The compounds have desirable pharmacokinetic properties, safety profiles and are effective in mouse infection models for biofilms at concentrations similar to or lower than current therapeutics. Based on these data, we suggest that modulation of the biofilm life-cycle, i.e. by inducing dispersal using NO, is a viable strategy for the treatment of chronic infections caused by P. aeruginosa.