Open AccessA dimerization-based fluorogenic dye-aptamer module for RNA imaging in live cells
Author(s) -
Farah Bouhedda,
Kyong Tkhe Fam,
Mayeul Collot,
Alexis Autour,
Stefano Marzi,
Andrey S. Klymchenko,
Michaël Ryckelynck
Publication year - 2019
Publication title -
nature chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.412
H-Index - 216
eISSN - 1552-4469
pISSN - 1552-4450
DOI - 10.1038/s41589-019-0381-8
Subject(s) - aptamer , rna , live cell imaging , fluorescence , sulforhodamine b , biophysics , chemistry , cell , microbiology and biotechnology , nanotechnology , biology , biochemistry , materials science , gene , physics , optics , cytotoxicity , in vitro
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.