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Red-Shifted Substrates for FAST Fluorogen-Activating Protein Based on the GFP-Like Chromophores |
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| Authors: | Dr. Natalia V. Povarova Snizhana O. Zaitseva Nadezhda S. Baleeva Alexander Yu. Smirnov Ivan N. Myasnyanko Marina B. Zagudaylova Dr. Nina G. Bozhanova Dmitriy A. Gorbachev Kseniya K. Malyshevskaya Alexey S. Gavrikov Dr. Alexander S. Mishin Dr. Mikhail S. Baranov |
| Affiliation: | 1. Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia;2. Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Center of Life Sciences, Skolkovo Institute of Science and Technology, Bolshoy Blvd 30, Moscow, 121205 Russia |
| 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. |
| Keywords: | chromophore fluorescence fluorescent probes fluorogen-activating protein green fluorescent protein |