Drude Irene, Vauléon Stéphanie, Müller Sabine
Ernst Moritz Arndt Universität Greifswald, Institut für Biochemie, Bioorganische Chemie, Felix Hausdorff Str. 4, 17487 Greifswald, Germany.
Biochem Biophys Res Commun. 2007 Nov 9;363(1):24-9. doi: 10.1016/j.bbrc.2007.08.135. Epub 2007 Aug 31.
Over the past two decades, the structure and mechanism of catalytic RNA have been extensively studied; now ribozymes are understood well enough to turn them into useful tools. After we have demonstrated the twin ribozyme mediated insertion of additional nucleotides into a predefined position of a suitable substrate RNA, we here show that a similar type of twin ribozyme is also capable of mediating the opposite reaction: the site-specific removal of nucleotides. In particular, we have designed a twin ribozyme that supports the deletion of four uridine residues from a given RNA substrate. This reaction is a kind of RNA recombination that in the specific context of gene therapy mimics, at the level of RNA, the correction of insertion mutations. As a result of the twin ribozyme driven reaction, 17% of substrate are converted into the four nucleotides shorter product RNA.
在过去二十年中，催化性RNA的结构和机制得到了广泛研究；如今对核酶的了解已足够深入，足以将它们转化为有用的工具。在我们证明了双体核酶介导将额外核苷酸插入合适底物RNA的预定位置后，我们在此表明，类似类型的双体核酶也能够介导相反的反应：核苷酸的位点特异性去除。具体而言，我们设计了一种双体核酶，它能支持从给定的RNA底物中删除四个尿苷残基。这种反应是一种RNA重组，在基因治疗模拟的特定背景下，在RNA水平上模拟插入突变的校正。由于双体核酶驱动的反应，17%的底物被转化为短四个核苷酸的产物RNA。
