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Circularly Permuted Fluorescent Protein-Based Indicators: History, Principles, and Classification.

Alexander I Kostyuk1,2, Aleksandra D Demidovich1, Daria A Kotova1

  • 1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.

International Journal of Molecular Sciences
|August 30, 2019
PubMed
Summary

Genetically encoded biosensors utilize circularly permuted fluorescent proteins (cpFPs) to monitor intracellular events. This review details their principles, history, and classification for biological process studies.

Keywords:
circularly permuted fluorescent proteinsgenetically encoded biosensors

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Genetically encoded biosensors based on fluorescent proteins (FPs) are vital for studying biological processes in living systems.
  • Circularly permuted FPs (cpFPs) enable monitoring of numerous intracellular events by fusing original termini and forming new ones near the chromophore.
  • cpFP structure offers greater mobility and spectral lability compared to native variants.

Purpose of the Study:

  • To review the fundamental principles behind genetically encoded biosensors utilizing circularly permuted fluorescent proteins (cpFPs).
  • To provide a historical overview of the development of these cpFP-based biosensors.
  • To present a comprehensive classification of currently available cpFP biosensors.

Main Methods:

  • Integration of cpFP into flexible regions or between interacting domains of sensory elements.
  • Leveraging conformational changes in sensory domains upon ligand interaction or cellular parameter shifts.
  • Transferring structural rearrangements to the cpFP, altering the chromophore environment and spectral characteristics.

Main Results:

  • cpFP-based biosensors facilitate the monitoring of diverse intracellular events through conformational changes.
  • The design principle involves linking cpFPs to sensory domains that respond to specific cellular parameters or ligands.
  • Spectral properties of cpFPs are modulated by their integration into biosensor frameworks.

Conclusions:

  • cpFP-based biosensors represent a powerful and versatile tool for live-cell imaging and biological research.
  • Understanding their design principles and classification is crucial for developing new biosensing applications.
  • This review consolidates knowledge on cpFP biosensors, aiding future advancements in the field.