Although several DNAzymes are known, their utility is limited by a narrow range of substrate specificity. Here, we report the isolation of two zinc-dependent DNAzymes, ZincDz1 and ZincDz2, which exhibit compact catalytic core sequences with highly versatile hydrolysis activity. They were selected through in vitro selection followed by deep sequencing analysis. Despite their sequence similarity, each DNAzyme showed different Zn²⁺-concentration and pH-dependent reaction profiles, and cleaved the target RNA sequences at different sites. Using various substrate RNA sequences, we found that the cleavage sequence specificity of ZincDz2 and its highly active mutant ZincDz2-v2 to be 5′-rN↓rNrPu-3′. Furthermore, we demonstrated that the designed ZincDz2 could cut microRNA miR-155 at three different sites. These DNAzymes could be useful in a broad range of applications in the fields of medicine and biotechnology.

尽管已知几种 DNA 酶，但它们的效用受到底物特异性范围狭窄的限制。在这里，我们报告了两种依赖锌的 DNAzymes ZincDz1 和 ZincDz2 的分离，它们表现出具有高度通用水解活性的紧凑催化核心序列。它们是通过体外选择和深度测序分析来选择的。尽管它们的序列相似，但每个 DNAzyme 表现出不同的 Zn ²⁺浓度和 pH 依赖性反应曲线，并在不同位点切割目标 RNA 序列。使用各种底物RNA序列，我们发现ZincDz2及其高活性突变体ZincDz2-v2的切割序列特异性为5'-rN↓rNrPu-3'。此外，我们证明了设计的 ZincDz2 可以在三个不同的位点切割 microRNA miR-155。这些 DNAzymes 可用于医学和生物技术领域的广泛应用。