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利用类似夹钳的三链体适体结构来切换 Taq 聚合酶的活性。

Switching the activity of Taq polymerase using clamp-like triplex aptamer structure.

机构信息

Key Laboratory of Image Information Processing and Intelligent Control of Education Ministry of China, School of Artificial Intelligence and Automation, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China.

College of Information Science and Technology, Shijiazhuang Tiedao University, Shijiazhuang, Hebei 050043, China.

出版信息

Nucleic Acids Res. 2020 Sep 4;48(15):8591-8600. doi: 10.1093/nar/gkaa581.

Abstract

In nature, allostery is the principal approach for regulating cellular processes and pathways. Inspired by nature, structure-switching aptamer-based nanodevices are widely used in artificial biotechnologies. However, the canonical aptamer structures in the nanodevices usually adopt a duplex form, which limits the flexibility and controllability. Here, a new regulating strategy based on a clamp-like triplex aptamer structure (CLTAS) was proposed for switching DNA polymerase activity via conformational changes. It was demonstrated that the polymerase activity could be regulated by either adjusting structure parameters or dynamic reactions including strand displacement or enzymatic digestion. Compared with the duplex aptamer structure, the CLTAS possesses programmability, excellent affinity and high discrimination efficiency. The CLTAS was successfully applied to distinguish single-base mismatches. The strategy expands the application scope of triplex structures and shows potential in biosensing and programmable nanomachines.

摘要

在自然界中,变构是调节细胞过程和途径的主要方法。受自然启发,基于结构切换的适体纳米器件广泛应用于人工生物技术中。然而,纳米器件中的典型适体结构通常采用双链体形式,这限制了其灵活性和可控性。在这里,提出了一种基于夹式三聚体适体结构 (CLTAS) 的新调控策略,通过构象变化来切换 DNA 聚合酶活性。结果表明,聚合酶活性可以通过调节结构参数或包括链置换或酶消化在内的动态反应来调节。与双链适体结构相比,CLTAS 具有可编程性、优异的亲和力和高的区分效率。CLTAS 成功地应用于区分单碱基错配。该策略扩展了三聚体结构的应用范围,并在生物传感和可编程纳米机器中显示出潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7a66/7470972/b9d320071092/gkaa581fig1.jpg

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