Generation of Transgenic Mice Expressing Cytosolic and Targeted FRET Biosensors for cAMP and cGMP.

Transgenic mice play a significant role in modern biomedical research. In addition to mechanistic studies of a specific gene and protein function, transgenic mice are used as an exciting tool for in vivo or in situ analysis of fluorescent biosensors, which are capable of directly reporting second messenger levels and biochemical processes in real time and living cells. In this chapter, we present a detailed protocol for the generation of plasmid vectors and transgenic mice ubiquitously or constitutively expressing cytosolic and targeted Förster resonance energy transfer (FRET)-based biosensors for the second messengers 3',5'-cyclic adenosine and guanosine monophosphates. These tools and techniques hold great potential for the analysis of second messenger dynamics in physiologically relevant systems.