Pseudomonas aeruginosa is a Gram-negative human pathogen that causes hard-to-eradicate infections in hospitalized, immunocompromised and cystic fibrosis patients. This ESKAPE pathogen is among the major model organisms for quorum sensing (QS) studies, with an average of about 200 publications per year focusing on P. aeruginosa QS in the last decade. The interest of the scientific community in P. aeruginosa QS can be explained by the fascinating complexity of this intercellular communication system and by its potential as a drug target. In fact, P. aeruginosa evolved complex QS machinery consisting of three interwoven QS systems, namely las, rhl and pqs, based on multiple signal molecules, cognate receptors, effectors and modulators. All together, these systems contribute to P. aeruginosa adaptation to the host environment, supporting QS as a promising target for the development of antivirulence drugs reducing P. aeruginosa pathogenicity. Besides complicating the understanding of their molecular mechanism of action and physiological role, the multiplicity of proteins contributing to P. aeruginosa QS provides a convenient source of molecular tools exploitable in biotechnological applications. This talk will recapitulate some studies about P. aeruginosa QS carried out by our group in the last years, from the characterization of basic molecular mechanisms contributing to the regulation of the las QS system, to the development of biosensors allowing identification of FDA-approved inhibitors targeting the pqs QS system, and to the generation of injectable synthetic minimal cells able to interface with natural cells through molecular components of the rhl QS system.