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水解性内切核酸酶核糖酶(HYER):核酸操作中的系统鉴定、表征及潜在应用

Hydrolytic endonucleolytic ribozyme (HYER): Systematic identification, characterization and potential application in nucleic acid manipulation.

作者信息

Liu Zi-Xian, Liu Jun-Jie Gogo

机构信息

Beijing Frontier Research Center for Biological Structure, Center of Synthetic and Systems Biology, State Key Laboratory of Membrane Biology, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing, P.R. China.

出版信息

Methods Enzymol. 2025;712:197-223. doi: 10.1016/bs.mie.2025.01.033. Epub 2025 Mar 6.

Abstract

Group II introns are transposable elements that can propagate in host genomes through the "copy and paste" mechanism. They usually comprise RNA and protein components for effective propagation. Recently, we found that some bacterial GII-C introns without protein components had multiple copies in their resident genomes, implicating their potential transposition activity. We demonstrated that some of these systems are active for hydrolytic DNA cleavage and proved their DNA manipulation capability in bacterial or mammalian cells. These introns are therefore named HYdrolytic Endonucleolytic Ribozymes (HYERs). Here, we provide a detailed protocol for the systematic identification and characterization of HYERs and present our perspectives on its potential application in nucleic acid manipulation.

摘要

II 类内含子是可通过“复制粘贴”机制在宿主基因组中传播的转座元件。它们通常包含 RNA 和蛋白质成分以实现有效传播。最近,我们发现一些不含蛋白质成分的细菌 GII-C 内含子在其宿主基因组中有多个拷贝,这暗示了它们潜在的转座活性。我们证明其中一些系统具有水解 DNA 切割活性,并证实了它们在细菌或哺乳动物细胞中的 DNA 操作能力。因此,这些内含子被命名为水解内切核酸酶核酶(HYERs)。在此,我们提供了一份用于系统鉴定和表征 HYERs 的详细方案,并阐述了我们对其在核酸操作中潜在应用的观点。

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