期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 56, 期 22, 页码 6142-6146出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201700054
关键词
biosensors; DNAzymes; feedback amplification; RNA cleavage; rolling-circle amplification
资金
- Natural Sciences and Engineering Research Council of Canada (NSERC)
- Ontario Ministry of Research and Innovation
- Canada Foundation for Innovation (CFI)
We report a signal amplification strategy termed DNAzyme feedback amplification (DFA) that takes advantage of rolling-circle amplification (RCA) and an RNA-cleaving DNAzyme (RCD). DFA employs two specially programmed DNA complexes, one composed of a primer and a circular template containing the antisense sequence of an RCD, and the other composed of the same circular template and an RNA-containing substrate for the RCD. RCA is initiated at the first complex to produce RCD elements that go on to cleave the substrate in the second complex. This cleavage event triggers the production of more input complexes for RCA. This reaction circuit continues autonomously, resulting in exponential DNA amplification. We demonstrate the versatility of this approach for biosensing through the design of DFA systems capable of detecting a microRNA sequence and a bacterium, with sensitivity improvements of 3-6 orders of magnitude over conventional methods.
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