3 years ago

A DNAzyme Feedback Amplification Strategy for Biosensing

A DNAzyme Feedback Amplification Strategy for Biosensing
Qiang Zhang, John D. Brennan, Yingfu Li, Dingran Chang, Jimmy Gu, Meng Liu
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. Molecular ping pong: Special DNA assemblies were designed to enable autonomous multistep cyclic actions by a DNA polymerase (Pol) and a DNAzyme (DZ) that can turn a limited number of molecular recognition events into large amounts of DNA amplicon for biosensing applications. Blue=circular DNA template, red=DNA primer, green=RNA-cleaving DNAzyme.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/anie.201700054

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