Takezawa Yusuke, Zhang Hanci, Mori Keita, Hu Lingyun, Shionoya Mitsuhiko
Department of Chemistry, Graduate School of Science, The University of Tokyo 7-3-1 Hongo, Bunkyo-ku Tokyo 113-0033 Japan
Chem Sci. 2024 Jan 20;15(7):2365-2370. doi: 10.1039/d3sc05042d. eCollection 2024 Feb 14.
A Cu-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-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-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, but enhanced 21-fold with the addition of Cu. Furthermore, the DNAzyme activity was turned on and off during the reaction by the addition and removal of Cu ions. Both ligase-mediated synthesis and Cu-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.
通过引入双面5-羧基尿嘧啶（caU）核碱基，开发了一种对铜有响应的变构脱氧核酶，该核碱基形成氢键结合的caU-A和金属介导的caU-Cu-caU碱基对。基于已知的RNA切割脱氧核酶对碱基序列进行了合理设计，以便caU修饰的脱氧核酶（caU-脱氧核酶）能够形成包含三个caU-A碱基对的催化无活性结构和具有三个caU-Cu-caU对的活性形式。caU-脱氧核酶是通过用标准DNA连接酶连接含caU的短片段合成的。caU-脱氧核酶在没有铜的情况下活性受到抑制，但添加铜后活性增强了21倍。此外，通过添加和去除铜离子，在反应过程中脱氧核酶的活性可以开启和关闭。连接酶介导的合成和铜依赖性变构调节均通过caU的双面碱基配对特性实现。本研究为设计刺激响应性DNA分子系统提供了一种新策略。
