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Optogenetic techniques have become widespread throughout the life-science community since first rising to prominence about a decade ago.1  Compared with conventional pharmacological and electrophysiological techniques, optical methods are relatively non-invasive and provide high spatiotemporal resolution. Furthermore, genetic introduction of optogenetic reporters, although requiring greater up-front investment of time and resources, permits precise targeting of biological events at the molecular level and can greatly facilitate experimental investigations in the long term. The combination of optical and genetic methods, for the first time in history, allows us to interrogate specific biological processes under close to native conditions.2,3 

Originally, the term “optogenetics” referred only to the optical control of electrical activity in neurons with microbial opsins expressed heterologously.1  The repertoire of optogenetic control has since been expanded with the development and use of a variety of optogenetic actuators.3–6  In contrast to optogenetic actuators that transduce light into biological stimuli, optogenetic reporters reveal biological events by producing detectable optical readouts due to changes in light absorbance or emission.7–9  Historically, the discovery and extensive engineering of genetically encoded light-emitting proteins, including fluorescent proteins (FPs)10,11  and luciferases,12  have triggered the development of genetically encoded reporters and now still provide the basis for constructing new optogenetic reporters. Advances in FP and bioluminescence technologies are covered in Chapter 3. This chapter focuses on the engineering and applications of FP-based optogenetic reporters.

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