Orthogonal fluorescent chemogenetic reporters for multicolor imaging

Alison G. Tebo, Benjamien Moeyaert, Marion Thauvin, Irene Carlon-Andres, Dorothea Böken, Michel Volovitch, Sergi Padilla-Parra, Peter Dedecker, Sophie Vriz, Arnaud Gautier*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

36 Citations (Scopus)

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. [Figure not available: see fulltext.].

Original languageEnglish
Pages (from-to)30-38
Number of pages9
JournalNature Chemical Biology
Volume17
Issue number1
DOIs
Publication statusPublished - Jan 2021

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