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In vitro Selection of Chemically Modified DNAzymes
ChemistryOpen ( IF 2.3 ) Pub Date : 2020-10-19 , DOI: 10.1002/open.202000134
Po-Jung Jimmy Huang 1 , Juewen Liu 1
Affiliation  

DNAzymes are in vitro selected DNA oligonucleotides with catalytic activities. RNA cleavage is one of the most extensively studied DNAzyme reactions. To expand the chemical functionality of DNA, various chemical modifications have been made during and after selection. In this review, we summarize examples of RNA‐cleaving DNAzymes and focus on those modifications introduced during in vitro selection. By incorporating various modified nucleotides via polymerase chain reaction (PCR) or primer extension, a few DNAzymes were obtained that can be specifically activated by metal ions such as Zn2+ and Hg2+. In addition, some modifications were introduced to mimic RNase A that can cleave RNA substrates in the absence of divalent metal ions. In addition, single modifications at the fixed regions of DNA libraries, especially at the cleavage junctions, have been tested, and examples of DNAzymes with phosphorothioate and histidine‐glycine modified tertiary amine were successfully obtained specific for Cu2+, Cd2+, Zn2+, and Ni2+. Labeling fluorophore/quencher pair right next to the cleavage junction was also used to obtain signaling DNAzymes for detecting various metal ions and cells. Furthermore, we reviewed work on the cleavage of 2′‐5′ linked RNA and L‐RNA substrates. Finally, applications of these modified DNAzymes as biosensors, RNases, and biochemical probes are briefly described with a few future research opportunities outlined at the end.

中文翻译:

化学修饰 DNAzyme 的体外选择

DNA酶是体外选择的具有催化活性的DNA寡核苷酸。RNA 切割是研究最广泛的 DNAzyme 反应之一。为了扩展 DNA 的化学功能,在选择期间和之后进行了各种化学修饰。在这篇综述中,我们总结了 RNA 切割 DNAzyme 的例子,并重点关注体外选择过程中引入的修饰。通过聚合酶链式反应(PCR)或引物延伸掺入各种修饰的核苷酸,获得了一些可以被Zn 2+和Hg 2+等金属离子特异性激活的DNA酶。此外,还引入了一些修饰来模拟 RNase A,可以在不存在二价金属离子的情况下切割 RNA 底物。此外,还测试了 DNA 文库固定区域,特别是裂解连接处的单一修饰,并成功获得了具有硫代磷酸酯和组氨酸-甘氨酸修饰叔胺的 DNAzyme 的实例,对 Cu 2+、Cd 2+、Zn具有特异性。 2+和 Ni 2+。紧邻裂解连接处的标记荧光团/猝灭剂对也用于获得用于检测各种金属离子和细胞的信号脱氧核糖核酸酶。此外,我们回顾了 2'‐5' 连接的 RNA 和 L-RNA 底物裂解的工作。最后,简要描述了这些修饰的 DNAzyme 作为生物传感器、RNA 酶和生化探针的应用,并在最后概述了一些未来的研究机会。
更新日期:2020-10-19
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