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

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DOI

  • Benjamin B. Kim, Stanford University
  • ,
  • Haodi Wu, Stanford University, University of Pittsburgh
  • ,
  • Yukun A. Hao, Stanford University
  • ,
  • Michael Pan, Stanford University
  • ,
  • Mariya Chavarha, Stanford University
  • ,
  • Yufeng Zhao, Stanford University
  • ,
  • Michael Westberg
  • François St-Pierre, Baylor College of Medicine
  • ,
  • Joseph C. Wu, Stanford University
  • ,
  • Michael Z. Lin, Stanford University

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.

OriginalsprogEngelsk
Artikelnummer3678
TidsskriftScientific Reports
Vol/bind12
Nummer1
Antal sider11
ISSN2045-2322
DOI
StatusUdgivet - 2022

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