Living-Cell MicroRNA Imaging with Self-Assembling Fragments of Fluorescent Protein-Mimic RNA Aptamer
| Autor: | Mei-Rong Cui, Xueli Zhao, Xue-Jiao Yang, Jing-Juan Xu, Ting-Ting Zhang, Yu Gu, Li-Juan Huang, Hong-Yuan Chen, Wei Zhao |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Fluid Flow and Transfer Processes
Messenger RNA Chemistry Process Chemistry and Technology Aptamer 010401 analytical chemistry RNA Bioengineering 02 engineering and technology Transfection Aptamers Nucleotide 021001 nanoscience & nanotechnology G-quadruplex Non-coding RNA 01 natural sciences Fluorescence 0104 chemical sciences Cell biology G-Quadruplexes MicroRNAs microRNA Nucleic acid RNA Messenger 0210 nano-technology Instrumentation |
| Zdroj: | ACS sensors. 6(6) |
| ISSN: | 2379-3694 |
| Popis: | As the cellular roles of RNA abundance continue to increase, there is an urgent need for the corresponding tools to elucidate native RNA functions and dynamics, especially those of short, low-abundance RNAs in live cells. Fluorescent RNA aptamers provide a useful strategy to create the RNA tag and biosensor devices. Corn, which binds with 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO), is a good candidate for the RNA tag because of its enhanced photostability and red-shifted spectrum. Herein, we report for the first time the utilization of Corn as a split aptamer system, combined with RNA-initiated fluorescence complementation (RIFC), for monitoring RNA self-assembly and sensing microRNA. In this platform, the 28-nt Corn was divided into two nonfunctional halves (named probe I and probe II), and an additional target RNA recognition and stem part was introduced in each probe. The target RNA can trigger the self-assembly reconstitution of the Corn's G-quadruplex scaffold for DFHO binding and turn-on fluorescence. These probes can be transfected stably into mammalian cells and deliver the light-up fluorescent response to microRNA-21 (miR-21). Significantly, the probes have good photostability, with minimal fluorescence loss after continuous irradiation, and can be used for imaging of miR-21 in living mammalian cells. The proposed method is universal and could be applied to the sensing of other tumor-associated RNAs, including messenger RNA and noncoding RNA, as well as for monitoring RNA/RNA interactions. The Corn-based splitting aptamers show promising potential in the real-time visualization and mechanistic analysis of nucleic acids. |
| Databáze: | OpenAIRE |
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