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Plug-and-Play Fluorophores Extend the Spectral Properties of Spinach
[Image: see text] Spinach and Spinach2 are RNA aptamers that can be used for the genetic encoding of fluorescent RNA. Spinach2 binds and activates the fluorescence of (Z)-4-(3,5-difluoro-4-hydroxybenzylidene)-1,2-dimethyl-1H-imidazol-5(4H)-one (DFHBI), allowing the dynamic localizations of Spinach2-...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2014
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3929357/ https://www.ncbi.nlm.nih.gov/pubmed/24393009 http://dx.doi.org/10.1021/ja410819x |
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author | Song, Wenjiao Strack, Rita L. Svensen, Nina Jaffrey, Samie R. |
author_facet | Song, Wenjiao Strack, Rita L. Svensen, Nina Jaffrey, Samie R. |
author_sort | Song, Wenjiao |
collection | PubMed |
description | [Image: see text] Spinach and Spinach2 are RNA aptamers that can be used for the genetic encoding of fluorescent RNA. Spinach2 binds and activates the fluorescence of (Z)-4-(3,5-difluoro-4-hydroxybenzylidene)-1,2-dimethyl-1H-imidazol-5(4H)-one (DFHBI), allowing the dynamic localizations of Spinach2-tagged RNAs to be imaged in live cells. The spectral properties of Spinach2 are limited by DFHBI, which produces fluorescence that is bluish-green and is not optimized for filters commonly used in fluorescence microscopes. Here we characterize the structural features that are required for fluorophore binding to Spinach2 and describe novel fluorophores that bind and are switched to a fluorescent state by Spinach2. These diverse Spinach2–fluorophore complexes exhibit fluorescence that is more compatible with existing microscopy filter sets and allows Spinach2-tagged constructs to be imaged with either GFP or YFP filter cubes. Thus, these “plug-and-play” fluorophores allow the spectral properties of Spinach2 to be altered on the basis of the specific spectral needs of the experiment. |
format | Online Article Text |
id | pubmed-3929357 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-39293572014-02-20 Plug-and-Play Fluorophores Extend the Spectral Properties of Spinach Song, Wenjiao Strack, Rita L. Svensen, Nina Jaffrey, Samie R. J Am Chem Soc [Image: see text] Spinach and Spinach2 are RNA aptamers that can be used for the genetic encoding of fluorescent RNA. Spinach2 binds and activates the fluorescence of (Z)-4-(3,5-difluoro-4-hydroxybenzylidene)-1,2-dimethyl-1H-imidazol-5(4H)-one (DFHBI), allowing the dynamic localizations of Spinach2-tagged RNAs to be imaged in live cells. The spectral properties of Spinach2 are limited by DFHBI, which produces fluorescence that is bluish-green and is not optimized for filters commonly used in fluorescence microscopes. Here we characterize the structural features that are required for fluorophore binding to Spinach2 and describe novel fluorophores that bind and are switched to a fluorescent state by Spinach2. These diverse Spinach2–fluorophore complexes exhibit fluorescence that is more compatible with existing microscopy filter sets and allows Spinach2-tagged constructs to be imaged with either GFP or YFP filter cubes. Thus, these “plug-and-play” fluorophores allow the spectral properties of Spinach2 to be altered on the basis of the specific spectral needs of the experiment. American Chemical Society 2014-01-06 2014-01-29 /pmc/articles/PMC3929357/ /pubmed/24393009 http://dx.doi.org/10.1021/ja410819x Text en Copyright © 2014 American Chemical Society Terms of Use (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) |
spellingShingle | Song, Wenjiao Strack, Rita L. Svensen, Nina Jaffrey, Samie R. Plug-and-Play Fluorophores Extend the Spectral Properties of Spinach |
title | Plug-and-Play
Fluorophores Extend the Spectral Properties
of Spinach |
title_full | Plug-and-Play
Fluorophores Extend the Spectral Properties
of Spinach |
title_fullStr | Plug-and-Play
Fluorophores Extend the Spectral Properties
of Spinach |
title_full_unstemmed | Plug-and-Play
Fluorophores Extend the Spectral Properties
of Spinach |
title_short | Plug-and-Play
Fluorophores Extend the Spectral Properties
of Spinach |
title_sort | plug-and-play
fluorophores extend the spectral properties
of spinach |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3929357/ https://www.ncbi.nlm.nih.gov/pubmed/24393009 http://dx.doi.org/10.1021/ja410819x |
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