A red fluorescent protein with improved monomericity enables ratiometric voltage imaging with ASAP3

A ratiometric genetically encoded voltage indicator (GEVI) would be desirable for tracking transmembrane voltage changes in the presence of sample motion. We performed combinatorial multi-site mutagenesis on a cyan-excitable red fluorescent protein to create the bright and monomeric mCyRFP3, which p...

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Autores principales: Kim, Benjamin B., Wu, Haodi, Hao, Yukun A., Pan, Michael, Chavarha, Mariya, Zhao, Yufeng, Westberg, Michael, St-Pierre, François, Wu, Joseph C., Lin, Michael Z.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8901740/
https://www.ncbi.nlm.nih.gov/pubmed/35256624
http://dx.doi.org/10.1038/s41598-022-07313-1
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author Kim, Benjamin B.
Wu, Haodi
Hao, Yukun A.
Pan, Michael
Chavarha, Mariya
Zhao, Yufeng
Westberg, Michael
St-Pierre, François
Wu, Joseph C.
Lin, Michael Z.
author_facet Kim, Benjamin B.
Wu, Haodi
Hao, Yukun A.
Pan, Michael
Chavarha, Mariya
Zhao, Yufeng
Westberg, Michael
St-Pierre, François
Wu, Joseph C.
Lin, Michael Z.
author_sort Kim, Benjamin B.
collection PubMed
description A ratiometric genetically encoded voltage indicator (GEVI) would be desirable for tracking transmembrane voltage changes in the presence of sample motion. We performed combinatorial multi-site mutagenesis on a cyan-excitable red fluorescent protein to create the bright and monomeric mCyRFP3, which proved to be uniquely non-perturbing when fused to the GEVI ASAP3. The green/red ratio from ASAP3-mCyRFP3 (ASAP3-R3) reported voltage while correcting for motion artifacts, allowing the visualization of membrane voltage changes in contracting cardiomyocytes and throughout the cell cycle of motile cells.
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spelling pubmed-89017402022-03-08 A red fluorescent protein with improved monomericity enables ratiometric voltage imaging with ASAP3 Kim, Benjamin B. Wu, Haodi Hao, Yukun A. Pan, Michael Chavarha, Mariya Zhao, Yufeng Westberg, Michael St-Pierre, François Wu, Joseph C. Lin, Michael Z. Sci Rep Article A ratiometric genetically encoded voltage indicator (GEVI) would be desirable for tracking transmembrane voltage changes in the presence of sample motion. We performed combinatorial multi-site mutagenesis on a cyan-excitable red fluorescent protein to create the bright and monomeric mCyRFP3, which proved to be uniquely non-perturbing when fused to the GEVI ASAP3. The green/red ratio from ASAP3-mCyRFP3 (ASAP3-R3) reported voltage while correcting for motion artifacts, allowing the visualization of membrane voltage changes in contracting cardiomyocytes and throughout the cell cycle of motile cells. Nature Publishing Group UK 2022-03-07 /pmc/articles/PMC8901740/ /pubmed/35256624 http://dx.doi.org/10.1038/s41598-022-07313-1 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Kim, Benjamin B.
Wu, Haodi
Hao, Yukun A.
Pan, Michael
Chavarha, Mariya
Zhao, Yufeng
Westberg, Michael
St-Pierre, François
Wu, Joseph C.
Lin, Michael Z.
A red fluorescent protein with improved monomericity enables ratiometric voltage imaging with ASAP3
title A red fluorescent protein with improved monomericity enables ratiometric voltage imaging with ASAP3
title_full A red fluorescent protein with improved monomericity enables ratiometric voltage imaging with ASAP3
title_fullStr A red fluorescent protein with improved monomericity enables ratiometric voltage imaging with ASAP3
title_full_unstemmed A red fluorescent protein with improved monomericity enables ratiometric voltage imaging with ASAP3
title_short A red fluorescent protein with improved monomericity enables ratiometric voltage imaging with ASAP3
title_sort red fluorescent protein with improved monomericity enables ratiometric voltage imaging with asap3
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8901740/
https://www.ncbi.nlm.nih.gov/pubmed/35256624
http://dx.doi.org/10.1038/s41598-022-07313-1
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