A dimerization-based fluorogenic dye-aptamer module for RNA imaging in live cells.
Fiche publication
Date publication
octobre 2019
Journal
Nature chemical biology
Auteurs
Membres identifiés du Cancéropôle Est :
Dr RYCKELYNCK Mickael , Dr KLYMCHENKO Andrey
Tous les auteurs :
Bouhedda F, Fam KT, Collot M, Autour A, Marzi S, Klymchenko A, Ryckelynck M
Lien Pubmed
Résumé
Live-cell imaging of RNA has remained a challenge because of the lack of naturally fluorescent RNAs. Recently developed RNA aptamers that can light-up small fluorogenic dyes could overcome this limitation, but they still suffer from poor brightness and photostability. Here, we propose the concept of a cell-permeable fluorogenic dimer of self-quenched sulforhodamine B dyes (Gemini-561) and the corresponding dimerized aptamer (o-Coral) that can drastically enhance performance of the current RNA imaging method. The improved brightness and photostability, together with high affinity of this complex, allowed direct fluorescence imaging in live mammalian cells of RNA polymerase III transcription products as well as messenger RNAs labeled with a single copy of the aptamer; that is, without tag multimerization. The developed fluorogenic module enables fast and sensitive detection of RNA inside live cells, while the proposed design concept opens the route to new generation of ultrabright RNA probes.
Référence
Nat. Chem. Biol.. 2019 Oct 21;: