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

Sci Rep. 2022 Mar 7;12(1):3678. doi: 10.1038/s41598-022-07313-1.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Diagnostic Imaging*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mutagenesis
  • Neurons* / metabolism
  • Red Fluorescent Protein

Substances

  • Luminescent Proteins