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Far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome

Ability to modulate fluorescence of optical probes can be used to enhance signal-to-noise ratio for imaging within highly autofluorescent environments, such as intact tissues and living organisms. Here we report two phytochrome-based photoactivatable near-infrared fluorescent proteins, named PAiRFP1...

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Autores principales: Piatkevich, Kiryl D., Subach, Fedor V., Verkhusha, Vladislav V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3749836/
https://www.ncbi.nlm.nih.gov/pubmed/23842578
http://dx.doi.org/10.1038/ncomms3153
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author Piatkevich, Kiryl D.
Subach, Fedor V.
Verkhusha, Vladislav V.
author_facet Piatkevich, Kiryl D.
Subach, Fedor V.
Verkhusha, Vladislav V.
author_sort Piatkevich, Kiryl D.
collection PubMed
description Ability to modulate fluorescence of optical probes can be used to enhance signal-to-noise ratio for imaging within highly autofluorescent environments, such as intact tissues and living organisms. Here we report two phytochrome-based photoactivatable near-infrared fluorescent proteins, named PAiRFP1 and PAiRFP2. PAiRFPs utilize heme-derived biliverdin, ubiquitous in mammalian tissues, as the chromophore. Initially weakly fluorescent PAiRFPs undergo photoconversion into a highly fluorescent state with excitation/emission at 690 nm/717 nm following a brief irradiation with far-red light. After photoactivation, PAiRFPs slowly revert back to initial state, enabling multiple photoactivation-relaxation cycles. Low-temperature optical spectroscopy reveals several intermediates involved in PAiRFP photocycles, which all differ from that of the bacteriophytochrome precursor. PAiRFPs can be photoactivated in a spatially selective manner in mouse tissues, and optical modulation of their fluorescence allows for substantial contrast enhancement, making PAiRFPs advantageous over permanently fluorescent probes for in vivo imaging conditions of high autofluorescence and low signal levels.
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spelling pubmed-37498362014-01-10 Far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome Piatkevich, Kiryl D. Subach, Fedor V. Verkhusha, Vladislav V. Nat Commun Article Ability to modulate fluorescence of optical probes can be used to enhance signal-to-noise ratio for imaging within highly autofluorescent environments, such as intact tissues and living organisms. Here we report two phytochrome-based photoactivatable near-infrared fluorescent proteins, named PAiRFP1 and PAiRFP2. PAiRFPs utilize heme-derived biliverdin, ubiquitous in mammalian tissues, as the chromophore. Initially weakly fluorescent PAiRFPs undergo photoconversion into a highly fluorescent state with excitation/emission at 690 nm/717 nm following a brief irradiation with far-red light. After photoactivation, PAiRFPs slowly revert back to initial state, enabling multiple photoactivation-relaxation cycles. Low-temperature optical spectroscopy reveals several intermediates involved in PAiRFP photocycles, which all differ from that of the bacteriophytochrome precursor. PAiRFPs can be photoactivated in a spatially selective manner in mouse tissues, and optical modulation of their fluorescence allows for substantial contrast enhancement, making PAiRFPs advantageous over permanently fluorescent probes for in vivo imaging conditions of high autofluorescence and low signal levels. 2013 /pmc/articles/PMC3749836/ /pubmed/23842578 http://dx.doi.org/10.1038/ncomms3153 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Piatkevich, Kiryl D.
Subach, Fedor V.
Verkhusha, Vladislav V.
Far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome
title Far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome
title_full Far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome
title_fullStr Far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome
title_full_unstemmed Far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome
title_short Far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome
title_sort far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3749836/
https://www.ncbi.nlm.nih.gov/pubmed/23842578
http://dx.doi.org/10.1038/ncomms3153
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