Publication Date
3-7-2022
Journal
Scientific Reports
DOI
10.1038/s41598-022-07313-1
PMID
35256624
PMCID
PMC8901740
PubMedCentral® Posted Date
3-7-2022
PubMedCentral® Full Text Version
Post-print
Published Open-Access
yes
Keywords
Diagnostic Imaging, Luminescent Proteins, Mutagenesis, Neurons, Red Fluorescent Protein, Fluorescence imaging, Protein design
Abstract
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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