Red‐Shifted Substrates for FAST Fluorogen‐Activating Protein Based on the GFP‐Like Chromophores

NV Povarova, SO Zaitseva, NS Baleeva… - … A European Journal, 2019 - Wiley Online Library
NV Povarova, SO Zaitseva, NS Baleeva, AY Smirnov, IN Myasnyanko, MB Zagudaylova…
Chemistry–A European Journal, 2019Wiley Online Library
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 …
Abstract
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.
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