Aptamer-Braked Multi-hairpin Cascade Circuits for Logic-Controlled Label-Free In Situ Bioimaging.,Analytical Chemistry

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Aptamer-Braked Multi-hairpin Cascade Circuits for Logic-Controlled Label-Free In Situ Bioimaging.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-06-29 , DOI: 10.1021/acs.analchem.0c00583
Zhijin Tian ₁ , Pai Peng ₁ , Huihui Wang ₁ , Jiao Zheng ₁ , Lili Shi ₁ , Tao Li ₁

Affiliation

As a common hairpin-based amplification strategy, catalytic-hairpin assembly (CHA) has been widely used to construct various DNA circuits for biosensing and imaging. However, the hairpin substrates can potentially react without catalysts and result in circuit leakage, which may be quite severe in a CHA reaction consisting of three or four hairpins due to the formation of stable three-/four-way junction product. To circumvent this problem, here we introduce a well-designed ATP aptamer as a DNA brake into a four-hairpin cascade circuit, where the triggering toehold is blocked by the aptamer brake and thus the circuit leakage decreases dramatically. Such an aptamer-braked DNA circuit is then employed to build an AND logic gate in response to multiple external stimuli in acidic cell membrane microenvironments. Induced by a bimolecular i-motif that binds thioflavin T (ThT), the dimerization of a four-way junction in situ assembled on the cell surface is accomplished, enabling the logic-controlled cell membrane imaging in a label-free manner. Our design would be applicable to other hairpin-based amplification strategies and may find more applications in the construction of multiresponsive DNA cascade circuits in complex living systems.

中文翻译：

适用于逻辑控制的无标记原位生物成像的适配臂多发夹级联电路。

作为基于发夹的常见扩增策略，催化发夹装配（CHA）已被广泛用于构建用于生物传感和成像的各种DNA电路。但是，发夹式底物可能会在没有催化剂的情况下发生反应并导致电路泄漏，由于形成稳定的三向/四向结产物，在由三个或四个发夹构成的CHA反应中这可能非常严重。为了解决这个问题，在这里我们将设计良好的ATP适配子作为DNA制动器引入到四发夹级联电路中，在该电路中，触发脚趾被适配子制动器阻挡，因此电路泄漏急剧减少。然后，将这种适配子激活的DNA电路用于响应酸性细胞膜微环境中的多个外部刺激来构建AND逻辑门。原位组装在细胞表面完成，从而实现了无标记方式的逻辑控制细胞膜成像。我们的设计将适用于其他基于发夹的扩增策略，并可能在复杂的生活系统中的多响应DNA级联电路的构建中找到更多的应用。

更新日期：2020-08-04

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