The signaling molecule nitric oxide (NO) acts as physiological activator of NO-sensitive guanylyl cyclase (NO-GC) in the cardiovascular, gastrointestinal and nervous systems. Two isoforms of NO-GC are known to exist on the protein level. The enzyme is a heterodimer consisting of an alpha (alpha(1) or alpha(2)) and a beta subunit (beta(1)). Strategies for the genomic deletion of either subunit have been developed in the recent years. Removal of one of the two isoforms by deletion of one of the alpha subunits allowed the investigation of the specific functions of the respective isoform. The deletion of the beta(1) subunit led to complete knock-out thus completely disrupting the NO/cGMP signaling cascade. The phenotypes of these KO mice have corroborated the already known physiological importance of the NO/cGMP cascade e.g. in the regulation of blood pressure, platelet inhibition, interneuronal communication; yet, they have also given hints to novel functions and mechanisms. In addition, mice lacking both NO-GC isoforms permitted the investigation of possible cGMP-independent signaling pathways of NO. As cell- and tissue-specific knock-out models are beginning to emerge, a more detailed analysis of the importance of the NO receptor in specific tissues will become possible.