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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Detalles Bibliográficos
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
Descripción
Sumario: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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