Powerful Amplification Cascades of FRET-Based Two-Layer Nonenzymatic Nucleic Acid Circuits

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• 作者：Ke Quan ; Jin Huang ; Xiaohai Yang ; Yanjing Yang ; Le Ying ; He Wang ; Nuli Xie ; Min Ou ; Kemin Wang
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：June 7, 2016
• 年：2016
• 卷：88
• 期：11
• 页码：5857-5864
• 全文大小：488K
• 年卷期：0
• ISSN：1520-6882

文摘

Nucleic acid circuits have played important roles in biological engineering and have increasingly attracted researchers’ attention. They are primarily based on nucleic acid hybridizations and strand displacement reactions between nucleic acid probes of different lengths. Signal amplification schemes that do not rely on protein enzyme show great potential in analytical applications. While the single amplification circuit often achieves linear amplification that may not meet the need for detection of target in a very small amount, it is very necessary to construct cascade circuits that allow for larger amplification of inputs. Herein, we have successfully engineered powerful amplification cascades of FRET-based two-layer nonenzymatic nucleic acid circuits, in which the outputs of catalyzed hairpin assembly (CHA) activate hybridization chain reactions (HCR) circuits to induce repeated hybridization, allowing real-time monitoring of self-assembly process by FRET signal. The cascades can yield 50000-fold signal amplification with the help of the well-designed and high-quality nucleic acid circuit amplifiers. Subsequently, with coupling of structure-switching aptamer, as low as 200 pM adenosine is detected in buffer, as well as in human serum. To our knowledge, we have for the first time realized real-time monitoring adaptation of HCR to CHA circuits and achieved amplified detection of nucleic acids and small molecules with relatively high sensitivity.