A Genetically Encodable System for Sequence‐Specific Detection of RNAs in Two Colors |
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| Authors: | Stefanie J Kellermann Prof Dr Andrea Rentmeister |
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| Affiliation: | 1. Department of Chemistry and Pharmacy, Institute of Biochemistry, University of Münster, Münster, Germany;2. Department of Chemistry, Institute for Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany;3. Cells-in-Motion Cluster of Excellence (EXC 1003–CiM), University of Münster, Münster, Germany |
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| Abstract: | Multicolor readout is an important feature of RNA detection techniques aiming at the investigation of RNA localization. Several detection methods have been developed, however they require either transfection of cells with the probe or prior tagging of the target RNA. We report a fully genetically encodable system for simultaneous detection of two RNAs by using green and yellow fluorescence based on tetramolecular fluorescence complementation (TetFC). To obtain yellow fluorescent protein (YFP), substitution T203Y was introduced into one of the three non‐fluorescent GFP fragments; this was fused to different variants of the Homo sapiens Pumilio homology domain. Using different sets of fusion proteins we were able to discriminate between two closely related target RNAs based on the fluorescence signals at the respective wavelengths. |
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| Keywords: | fluorescent probes GFP protein engineering Pumilio homology domain RNA recognition |
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