Kiliszek Agnieszka, Blaszczyk Leszek, Kierzek Ryszard, Rypniewski Wojciech
Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznan, Poland.
Nucleic Acids Res. 2017 Jun 2;45(10):e92. doi: 10.1093/nar/gkx122.
An RNA hairpin is an essential structural element of RNA. Hairpins play crucial roles in gene expression and intermolecular recognition but are also involved in the pathogenesis of some congenital diseases. Structural studies of the hairpin motifs are impeded by their thermodynamic instability, as they tend to unfold to form duplexes, especially at high concentrations required for crystallography or nuclear magnetic resonance spectroscopy. We have elaborated techniques to stabilize the RNA hairpins by linking the free ends of the RNA strand at the base of the hairpin stem. One method involves stilbene diether or hexaethylene glycol linkers and circularization by T4 RNA ligase. Another method uses click chemistry to stitch the RNA ends with a triazole linker. Both techniques are efficient and easy to perform. They should be useful in making stable, biologically relevant RNA constructs for structural studies.
RNA发夹是RNA的一种重要结构元件。发夹在基因表达和分子间识别中发挥着关键作用，但也参与了一些先天性疾病的发病机制。由于其热力学不稳定性，发夹基序的结构研究受到阻碍，因为它们倾向于展开形成双链体，尤其是在晶体学或核磁共振光谱所需的高浓度下。我们已经开发出通过连接发夹茎底部RNA链的自由末端来稳定RNA发夹的技术。一种方法涉及芪二醚或六甘醇连接体，并通过T4 RNA连接酶进行环化。另一种方法使用点击化学用三唑连接体连接RNA末端。这两种技术都高效且易于操作。它们在制备用于结构研究的稳定的、具有生物学相关性的RNA构建体方面应该是有用的。
