A multicolor riboswitch-based platform for imaging of RNA in live mammalian cells
| Autor: | Jacob T. Polaski, Sheng-Ting Hung, Ralph Jimenez, Amy E. Palmer, Joshua Wheeler, Robert T. Batey, Dilara Batan, Mikołaj Chromiński, Esther Braselmann, Zachariah E Holmes, Roy Parker, Aleksandra J. Wierzba, Dorota Gryko |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Riboswitch Aptamer Green Fluorescent Proteins Color Computational biology 010402 general chemistry 01 natural sciences Fluorescence Article 03 medical and health sciences Cell Line Tumor RNA Small Nuclear Escherichia coli Animals Humans Nucleotide Molecular Biology Fluorescent Dyes chemistry.chemical_classification Messenger RNA Chemistry HEK 293 cells RNA Cell Biology Aptamers Nucleotide Non-coding RNA 0104 chemical sciences 030104 developmental biology HEK293 Cells Cobalamin riboswitch Microscopy Fluorescence HeLa Cells Plasmids |
| Zdroj: | Nature chemical biology |
| ISSN: | 1552-4469 |
| Popis: | RNAs directly regulate a vast array of cellular processes, emphasizing the need for robust approaches to fluorescently label and track RNAs in living cells. Here, we develop an RNA imaging platform using the cobalamin riboswitch as an RNA tag and a series of probes containing cobalamin as a fluorescence quencher. This highly modular 'Riboglow' platform leverages different colored fluorescent dyes, linkers and riboswitch RNA tags to elicit fluorescence turn-on upon binding RNA. We demonstrate the ability of two different Riboglow probes to track mRNA and small noncoding RNA in live mammalian cells. A side-by-side comparison revealed that Riboglow outperformed the dye-binding aptamer Broccoli and performed on par with the gold standard RNA imaging system, the MS2-fluorescent protein system, while featuring a much smaller RNA tag. Together, the versatility of the Riboglow platform and ability to track diverse RNAs suggest broad applicability for a variety of imaging approaches. |
| Databáze: | OpenAIRE |
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