Abstract
Fluorescent biosensors to detect the bona fide events of biologically important molecules in living cells are increasingly demanded in the field of molecular cell biology. Recent advances in the development of fluorescent biosensors have made an outstanding contribution to elucidating not only the roles of individual biomolecules, but also the dynamic intracellular relationships between these molecules. However, rational design strategies of fluorescent biosensors are not as mature as they look. An insatiable request for the establishment of a more universal and versatile strategy continues to provide an attractive alternative, so-called modular strategy, which permits facile preparation of biosensors with tailored characteristics by a simple combination of a receptor and a signal transducer. This review describes an overview of the progress in design strategies of fluorescent biosensors, such as auto-fluorescent protein-based biosensors, protein-based biosensors covalently modified with synthetic fluorophores, and signaling aptamers, and highlights the insight into how a given receptor is converted to a fluorescent biosensor. Furthermore, we will demonstrate a significance of the modular strategy for the sensor design.
Highlights
Molecular tools for shedding light on the complex interplay between macromolecules, signaling molecules, and biologically important ions inside the cells play a central role in molecular and cell biology
Various kinds of fluorescent biosensors constructed by synthetic receptors [6,7,8,9,10,11,12,13] and biological macromolecular receptors such as proteins [5,14] and aptamers [15] have been reported to date
This review presents a brief survey of a variety of design strategies for fluorescent biosensors with an emphasis on the guiding principle
Summary
Molecular tools for shedding light on the complex interplay between macromolecules, signaling molecules, and biologically important ions inside the cells play a central role in molecular and cell biology. The design of analyte-sensitive sensors was based on AFP variants, whose fluorescent properties were directly affected by the interaction between a target molecule and a chromophore moiety in AFP. As well as for small organic molecules such as ATP [44], cAMP [45,46,47,48], cGMP [49,50,51], tryptophan [52], glutamate [53,54], and inositol phosphates [55,56] have been reported based on this strategy This type of sensors could be classified in three categories; (1) dual AFP-fused fluorescent resonance energy transfer (FRET)-based biosensors, (2) single circularly permuted (cp) AFP-based biosensors, and (3). We will describe these strategies in more detail, highlighting the insights from the practical standpoint
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