Ligase-mediated synthesis of CuII-responsive allosteric DNAzyme with bifacial 5-carboxyuracil nucleobases,Chemical Science

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Ligase-mediated synthesis of CuII-responsive allosteric DNAzyme with bifacial 5-carboxyuracil nucleobases
Chemical Science ( IF 7.6 ) Pub Date : 2024-01-20 , DOI: 10.1039/d3sc05042d
Yusuke Takezawa ₁ , Hanci Zhang ₁ , Keita Mori ₁ , Lingyun Hu ₁ , Mitsuhiko Shionoya ₁

Affiliation

A Cu^II-responsive allosteric DNAzyme has been developed by introducing bifacial 5-carboxyuracil (caU) nucleobases, which form both hydrogen-bonded caU–A and metal-mediated caU–Cu^II–caU base pairs. The base sequence was logically designed based on a known RNA-cleaving DNAzyme so that the caU-modified DNAzyme (caU-DNAzyme) can form a catalytically inactive structure containing three caU–A base pairs and an active form with three caU–Cu^II–caU pairs. The caU-DNAzyme was synthesized by joining short caU-containing fragments with a standard DNA ligase. The activity of caU-DNAzyme was suppressed without Cu^II, but enhanced 21-fold with the addition of Cu^II. Furthermore, the DNAzyme activity was turned on and off during the reaction by the addition and removal of Cu^II ions. Both ligase-mediated synthesis and Cu^II-dependent allosteric regulation were achieved by the bifacial base pairing properties of caU. This study provides a new strategy for designing stimuli-responsive DNA molecular systems.

中文翻译：

连接酶介导的 CuII 反应性变构 DNA 酶与双面 5-羧基尿嘧啶核碱基的合成

通过引入双面 5-羧基尿嘧啶（caU）核碱基开发了一种 Cu^II 反应性变构 DNA 酶，这些核碱基形成氢键 caU-A 和金属介导的 caU-Cu^II-caU 碱基对。碱基序列是基于已知的 RNA 裂解 DNA 酶进行逻辑设计的，因此 caU 修饰的 DNA 酶（caU-DNAzyme）可以形成包含三个 caU-A 碱基对和一个具有三个 caU-Cu^II-caU 对的活性形式的催化失活结构。通过将含有 caU 的短片段与标准 DNA 连接酶连接来合成 caU-DNA酶。caU-DNAzyme 的活性在没有 Cu^II 的情况下受到抑制，但随着 Cu^II 的加入而增强 21 倍。此外，在反应过程中，通过添加和去除 Cu^II 离子来打开和关闭 DNAzyme 活性。连接酶介导的合成和 Cu^II 依赖性变构调节都是通过 caU 的双面碱基配对特性实现的。本研究为设计刺激响应型 DNA 分子系统提供了一种新策略。

更新日期：2024-01-20

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