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

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Powerful Amplification Cascades of FRET-Based Two-Layer Nonenzymatic Nucleic Acid Circuits
Analytical Chemistry ( IF 6.7 ) Pub Date : 2016-05-10 00:00:00 , DOI: 10.1021/acs.analchem.6b00609
Ke Quan ₁ , Jin Huang ₁ , Xiaohai Yang ₁ , Yanjing Yang ₁ , Le Ying ₁ , He Wang ₁ , Nuli Xie ₁ , Min Ou ₁ , Kemin Wang ₁

Affiliation

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.

中文翻译：

基于FRET的两层非酶核酸回路的强大扩增级联

核酸回路在生物工程中起着重要的作用，并越来越引起研究人员的关注。它们主要基于不同长度的核酸探针之间的核酸杂交和链置换反应。不依赖于蛋白酶的信号放大方案在分析应用中显示出巨大的潜力。虽然单个放大电路通常会实现线性放大，可能无法满足少量检测目标的需要，但非常有必要构建允许较大输入放大的级联电路。在这里，我们已经成功地设计了基于FRET的两层非酶核酸电路的强大扩增级联，其中催化发夹装配（CHA）的输出激活杂交链反应（HCR）电路以诱导重复杂交，从而允许通过FRET信号实时监控自组装过程。借助精心设计的高质量核酸电路放大器，级联可以产生50000倍的信号放大。随后，结合结构转换适体，在缓冲液以及人血清中检测到低至200 pM的腺苷。据我们所知，我们首次实现了HCR对CHA电路的实时监控适应性，并以相对较高的灵敏度实现了核酸和小分子的扩增检测。借助精心设计的高质量核酸电路放大器，级联可以产生50000倍的信号放大。随后，结合结构转换适体，在缓冲液以及人血清中检测到低至200 pM的腺苷。据我们所知，我们首次实现了HCR对CHA电路的实时监控适应性，并以相对较高的灵敏度实现了核酸和小分子的扩增检测。借助精心设计的高质量核酸电路放大器，级联可以产生50000倍的信号放大。随后，结合结构转换适体，在缓冲液以及人血清中检测到低至200 pM的腺苷。据我们所知，我们首次实现了HCR对CHA电路的实时监控适配，并以相对较高的灵敏度实现了核酸和小分子的放大检测。

更新日期：2016-05-10

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