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Isolation of a site-specifically modified RNA from an unmodified transcript - art. no. e21

机译:从未修饰的转录本中分离位点特异性修饰的RNA-技术没有。 e21

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摘要

Natural RNAs contain many base modifications that have specific biological functions. The ability to functionally dissect individual modifications is facilitated by the identification and cloning of enzymes responsible for these modifications, but is hindered by the difficulty of isolating site-specifically modified RNAs away from unmodified transcripts. Using the m1G37 and m1A58 methyl modifications of tRNA as two examples, we demonstrate that non-pairing base modifications protect RNAs against the DNA-directed RNase H cleavage. This provide a new approach to obtain homogeneous RNAs with site-specific base modifications that are suitable for biochemical and functional studies.
机译:天然RNA包含许多具有特定生物学功能的碱基修饰。鉴定和克隆负责这些修饰的酶有助于在功能上分解单个修饰的能力,但由于难以将位点特异性修饰的RNA与未修饰的转录本分开而受到阻碍。使用tRNA的m1G37和m1A58甲基修饰作为两个例子,我们证明了非配对碱基修饰可保护RNA免受DNA定向的RNase H切割。这提供了一种获得具有适用于生化和功能研究的位点特异性碱基修饰的均质RNA的新方法。

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