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Chemistry. 2018 Mar 26;24(18):4473-4479. doi: 10.1002/chem.201705338. Epub 2018 Feb 1.

DNAzyme Feedback Amplification: Relaying Molecular Recognition to Exponential DNA Amplification.

Author information

1
School of Environmental Science and Technology, Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education), Dalian University of Technology, Dalian, 116024, P. R. China.
2
Department of Biochemistry and Biomedical Sciences and Chemistry & Chemical Biology, McMaster University, 1280 Main Street West, Hamilton, ON, L8S 4K1, Canada.
3
Biointerfaces Institute, McMaster University, 1280 Main Street West, Hamilton, ON, L8S 4L8, Canada.

Abstract

Technologies capable of linking DNA amplification to molecular recognition are very desirable for ultrasensitive biosensing applications. We have developed a simple but powerful isothermal DNA amplification method, termed DNAzyme feedback amplification (DFA), that is capable of relaying molecular recognition to exponential DNA amplification. The method incorporates both an RNA-cleaving DNAzyme (RCD) and rolling circle amplification (RCA) carried out by a special DNA polymerase using a circular DNA template. DFA begins with a stimulus-dependent RCA reaction, producing tandemly linked RCDs in long-chain DNA products. These RCDs cleave an RNA-containing DNA sequence to form additional primers that hybridize to the circular DNA molecule, giving rise to DNA assemblies that act as the new inputs for RCA. The RCA reaction and the cleavage event keep on feeding each other autonomously, resulting in exponential growth of repetitive DNA sequences that can be easily detected. This method can be used for the detection of both nucleic acid based targets and non-nucleic acid analytes. In this article, we discuss the conceptual framework of the feedback amplification approach, the essential features of this method as well as remaining challenges and possible solutions.

KEYWORDS:

DNA; RNA cleavage; biosensors; exponential amplification; rolling circle amplification

PMID:
29240289
DOI:
10.1002/chem.201705338
[Indexed for MEDLINE]

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