Development of Novel Cellular Imaging Tools Using Protein Engineering

Protein-based imaging probes provide the power to detect the location and activity of molecular events. Using protein engineering and directed evolution, genetically encoded protein modules have been developed to image live cells. In this chapter, we first introduce the protein engineering and development of various fluorescent proteins and their biosensor derivatives. We then discuss the engineering of binding proteins, i.e. antibody and non-antibody molecular scaffolds, which can be conjugated with various detection modules to image molecules in cells and target specific antigens in vivo . These developed fluorescent proteins and protein motifs are further introduced in applications related to molecular and cellular imaging of signaling events. Lastly, we discuss the applications of these tools for diagnostics and therapeutics via high throughput drug screening. In summary, protein engineering has driven the development of numerous molecular imaging probes for a wide range of applications, spanning from basic biology research to disease diagnosis and treatment.