Fluorescent proteins as genetically encoded FRET biosensors in life sciences.
Fluorescence- or Förster resonance energy transfer (FRET) is a measurable physical energy transfer phenomenon between appropriate chromophores, when they are in sufficient proximity, usually within 10 nm. This feature has made them incredibly useful tools for many biomedical studies on molecular interactions. Furthermore, this principle is increasingly exploited for the design of biosensors, where two chromophores are linked with a sensory domain controlling their distance and thus the degree of FRET. The versatility of these FRET-biosensors made it possible to assess a vast amount of biological variables in a fast and standardized manner, allowing not only high-throughput studies but also sub-cellular measurements of biological processes. In this review, we aim at giving an overview over the recent advances in genetically encoded, fluorescent-protein based FRET-biosensors, as these represent the largest and most vividly growing group of FRET-based sensors. For easy understanding, we are grouping them into four categories, depending on their molecular mechanism. These are based on: (a) cleavage; (b) conformational-change; (c) mechanical force and (d) changes in the micro-environment. We also address the many issues and considerations that come with the development of FRET-based biosensors, as well as the possibilities that are available to measure them.
Institute for Vascular Biology and Thrombosis Research, Medical University Vienna, Schwarzspanierstraße17, Vienna A-1090, Austria. bernhard.hochreiter@meduniwien.ac.at.
Bernhard
B
Hochreiter
Institute for Vascular Biology and Thrombosis Research, Medical University Vienna, Schwarzspanierstraße17, Vienna A-1090, Austria. alanpardogarcia@gmail.com.
Alan Pardo
AP
Garcia
Institute for Vascular Biology and Thrombosis Research, Medical University Vienna, Schwarzspanierstraße17, Vienna A-1090, Austria. johannes.schmid@meduniwien.ac.at.
Johannes A
JA
Schmid
D015374:Biosensing Techniques / Q000295:instrumentation*; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation*; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D008164:Luminescent Proteins / Q000032:analysis / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
D005456:Fluorescent Dyes; D008164:Luminescent Proteins
FRET; biosensors; fluorescence; imaging