期刊
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
卷 136, 期 52, 页码 17942-17945出版社
AMER CHEMICAL SOC
DOI: 10.1021/ja510839r
关键词
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资金
- University of California, San Diego
- NIH [K01EB010078, GM08326]
- American Cancer Society Moores Cancer Center, University of California, San Diego [70-002]
- Eugene V. Cota-Robles Fellowship
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.
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