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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Cheng, Jianjun Wu, Peiwen Zheng, Nan Mcghee, Claire E. Lu, Yi Chen, Lu Hwang, Kevin Torabi, Seyed-fakhreddin |
| Description | Author Affiliation: Torabi SF ( Departments of Biochemistry.); Wu P ( Departments of Biochemistry.); McGhee CE ( Chemistry, and.); Chen L ( Departments of Biochemistry.); Hwang K ( Chemistry, and.); Zheng N ( Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801.); Cheng J ( Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801.); Lu Y ( Departments of Biochemistry, Chemistry, and Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801 yi-lu@illinois.edu.); |
| Abstract | Over the past two decades, enormous progress has been made in designing fluorescent sensors or probes for divalent metal ions. In contrast, the development of fluorescent sensors for monovalent metal ions, such as sodium $(Na^{+}),$ has remained underdeveloped, even though $Na^{+}$ is one the most abundant metal ions in biological systems and plays a critical role in many biological processes. Here, we report the in vitro selection of the first (to our knowledge) $Na^{+}-specific,$ RNA-cleaving deoxyribozyme (DNAzyme) with a fast catalytic rate [observed rate constant $(k_{obs})$ ∼0.1 $min^{−1}],$ and the transformation of this DNAzyme into a fluorescent sensor for $Na^{+}$ by labeling the enzyme strand with a quencher at the 3′ end, and the DNA substrate strand with a fluorophore and a quencher at the 5′ and 3′ ends, respectively. The presence of $Na^{+}$ catalyzed cleavage of the substrate strand at an internal ribonucleotide adenosine (rA) site, resulting in release of the fluorophore from its quenchers and thus a significant increase in fluorescence signal. The sensor displays a remarkable selectivity (>10,000-fold) for $Na^{+}$ over competing metal ions and has a detection limit of 135 µM (3.1 ppm). Furthermore, we demonstrate that this DNAzyme-based sensor can readily enter cells with the aid of α-helical cationic polypeptides. Finally, by protecting the cleavage site of the $Na^{+}-specific$ DNAzyme with a photolabile o-nitrobenzyl group, we achieved controlled activation of the sensor after DNAzyme delivery into cells. Together, these results demonstrate that such a DNAzyme-based sensor provides a promising platform for detection and quantification of $Na^{+}$ in living cells. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 19 |
| Volume Number | 112 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2015-05-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Biosensing Techniques DNA, Catalytic Chemistry Fluorescent Dyes Sodium Catalysis Cations HeLa Cells Ions Microscopy, Confocal Nucleic Acids Peptides Potassium Protein Structure, Secondary RNA Spectrometry, Fluorescence Research Support, N.I.H., Extramural Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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