Protein biosensors based on the principle of fluorescence resonance energy transfer for monitoring cellular dynamics

Abstract

Genetically-coded, fluorescence resonance energy transfer (FRET) biosensors are widely used to study molecular events from single cells to whole organisms. They are unique among biosensors because of their spontaneous fluorescence and targeting specificity to both organelles and tissues. In this review, we discuss the theoretical basis of FRET with a focus on key parameters responsible for designing FRET biosensors that have the highest sensitivity. Next, we discuss recent applications that are grouped into four common biosensor design patterns--intermolecular FRET, intramolecular FRET, FRET from substrate cleavage and FRET using multiple colour fluorescent proteins. Lastly, we discuss recent progress in creating fluorescent proteins suitable for FRET purposes. Together these advances in the development of FRET biosensors are beginning to unravel the interconnected and intricate signalling processes as they are occurring in living cells and organisms.

Live-cell microscopy – tips and tools

“…The field of biosensors is growing rapidly (Hodgson et al, 2008;Li et al, 2006). Some of the many sensors that are available include ratiometric Ca 2+ sensors such as Fura-2AM (Invitrogen) or FRETbased FPs such as Premo-Cameleon (Invitrogen).…”

Fluorescent Protein Tracking and Detection: Fluorescent Protein Structure and Color Variants

“…(Rizzo et al 2009) provides an introduction to applications of FPs in live cells and tips on how FPs can be used with success in live cell imaging. Several examples of use of FPs for intracellular monitoring are described in Lalonde et al (2005), Li et al (2006), and Wang et al (2008).…”

Chemical Biosensors Based on Proteins Involved in Biomineralization Processes

“…However, there is the possibility of using them for monitoring biomolecules during diagnostic tests in different clinical areas (Chien et al, 2009;Cosnier, 1999;Navratilova et al, 2006). Recently, the development of a protein biosensor used to detect a specific class of antibiotic or any other biological important species have been reported elsewhere (Amine & Palleschi, 2004;Li et al, 2006;Mechler et al, 2006). Most of the proteins, which have been used for these types of structural and biomedical research, need to be in a higher degree of purity.…”