Red-Shifted Substrates for FAST Fluorogen-Activating Protein Based on the GFP-Like Chromophores.
Natalia V PovarovaSnizhana O ZaitsevaNadezhda S BaleevaAlexander Yu SmirnovIvan N MyasnyankoMarina B ZagudaylovaNina G BozhanovaDmitriy A GorbachevKseniya K MalyshevskayaAlexey S GavrikovAlexander S MishinMikhail S BaranovPublished in: Chemistry (Weinheim an der Bergstrasse, Germany) (2019)
A genetically encoded fluorescent tag for live cell microscopy is presented. This tag is composed of previously published fluorogen-activating protein FAST and a novel fluorogenic derivative of green fluorescent protein (GFP)-like chromophore with red fluorescence. The reversible binding of the novel fluorogen and FAST is accompanied by three orders of magnitude increase in red fluorescence (580-650 nm). The proposed dye instantly stains target cellular proteins fused with FAST, washes out in a minute timescale, and exhibits higher photostability of the fluorescence signal in confocal and widefield microscopy, in contrast with previously published fluorogen:FAST complexes.