| Autor: |
Wu H; Department of Chemistry and Biochemistry, University of California, San Diego , La Jolla, California 92093, United States., Cisneros BT, Cole CM, Devaraj NK |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of the American Chemical Society [J Am Chem Soc] 2014 Dec 31; Vol. 136 (52), pp. 17942-5. Date of Electronic Publication: 2014 Dec 19. |
| DOI: |
10.1021/ja510839r |
| Abstrakt: |
Tetrazine ligations have proven to be a powerful bioorthogonal technique for the detection of many labeled biomolecules, but the ligating nature of these reactions can limit reaction turnover in templated chemistry. We have developed a transfer reaction between 7-azabenzonorbornadiene derivatives and fluorogenic tetrazines that facilitates turnover amplification of the fluorogenic response in nucleic acid-templated reactions. Fluorogenic tetrazine-mediated transfer (TMT) reaction probes can be used to detect DNA and microRNA (miRNA) templates to 0.5 and 5 pM concentrations, respectively. The endogenous oncogenic miRNA target mir-21 could be detected in crude cell lysates and detected by imaging in live cells. Remarkably, the technique is also able to differentiate between miRNA templates bearing a single mismatch with high signal to background. We imagine that TMT reactions could find wide application for amplified fluorescent detection of clinically relevant nucleic acid templates. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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