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PLoS One. 2017 Sep 1;12(9):e0184225. doi: 10.1371/journal.pone.0184225. eCollection 2017.

Insertion of the voltage-sensitive domain into circularly permuted red fluorescent protein as a design for genetically encoded voltage sensor.

Author information

1
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russian Federation.
2
Institute of Higher Nervous Activity and Neurophysiology, Moscow, Russian Federation.
3
Center for Functional Connectomics, Korea Institute of Science & Technology, Seoul, Korea.
4
Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
5
Pirogov Russian National Research Medical University, Moscow, Russia.
6
Nizhny Novgorod State Medical Academy, Nizhny Novgorod, Russia.

Abstract

Visualization of electrical activity in living cells represents an important challenge in context of basic neurophysiological studies. Here we report a new voltage sensitive fluorescent indicator which response could be detected by fluorescence monitoring in a single red channel. To the best of our knowledge, this is the first fluorescent protein-based voltage sensor which uses insertion-into-circular permutant topology to provide an efficient interaction between sensitive and reporter domains. Its fluorescent core originates from red fluorescent protein (FP) FusionRed, which has optimal spectral characteristics to be used in whole body imaging techniques. Indicators using the same domain topology could become a new perspective for the FP-based voltage sensors that are traditionally based on Förster resonance energy transfer (FRET).

PMID:
28863184
PMCID:
PMC5580962
DOI:
10.1371/journal.pone.0184225
[Indexed for MEDLINE]
Free PMC Article

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