Orthogonal fluorescent chemogenetic reporters for multicolor imaging.
Nat Chem Biol
; 17(1): 30-38, 2021 01.
Article
em En
| MEDLINE
| ID: mdl-32778846
ABSTRACT
Spectrally separated fluorophores allow the observation of multiple targets simultaneously inside living cells, leading to a deeper understanding of the molecular interplay that regulates cell function and fate. Chemogenetic systems combining a tag and a synthetic fluorophore provide certain advantages over fluorescent proteins since there is no requirement for chromophore maturation. Here, we present the engineering of a set of spectrally orthogonal fluorogen-activating tags based on the fluorescence-activating and absorption shifting tag (FAST) that are compatible with two-color, live-cell imaging. The resulting tags, greenFAST and redFAST, demonstrate orthogonality not only in their fluorogen recognition capabilities, but also in their one- and two-photon absorption profiles. This pair of orthogonal tags allowed the creation of a two-color cell cycle sensor capable of detecting very short, early cell cycles in zebrafish development and the development of split complementation systems capable of detecting multiple protein-protein interactions by live-cell fluorescence microscopy.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Plasmídeos
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Coloração e Rotulagem
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Técnicas Biossensoriais
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Imagem Óptica
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Corantes Fluorescentes
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Biologia Molecular
Limite:
Animals
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article