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Abstract：
A variety of enzymes have been found to interact with double-stranded RNA (dsRNA) in order to carry out its functions. We have endeavored to prepare the covalently crosslinked native-like duplex RNA, which could be useful for biochemical studies and RNA nanotechnology. In this study, the interstrand covalently linked duplex RNA was formed by a crosslinking reaction between vinylpurine (VP) and the target cytosine or uracil in RNA. We measured melting temperatures and CD spectra to identify the properties of the VP crosslinked duplex RNA. The crosslinking formation increased the thermodynamic stability without disturbing the natural conformation of dsRNA. In addition, a competitive binding experiment with the duplex RNA binding enzyme, ADAR2, showed the crosslinked dsRNA bound the protein with nearly the same binding affinity as the natural dsRNA, confirming that it has finely preserved the natural traits of duplex RNA.
摘要：
已经发现多种酶与双链RNA(dsRNA)相互作用以执行其功能。我们努力制备共价交联的天然样双链体RNA，这可能对生化研究和纳米技术有用。在这项研究中,通过乙烯基嘌呤（VP）与RNA中的靶胞嘧啶或尿嘧啶之间的交联反应形成链间共价连接的双链体RNA。我们测量了解链温度和CD光谱以鉴定VP交联双链体RNA的性质。交联形成增加了热力学稳定性而不干扰dsRNA的天然构象。此外,双链体RNA结合酶的竞争性结合实验，ADAR2显示交联的dsRNA以与天然dsRNA几乎相同的结合亲和力结合蛋白质，证实它很好地保留了双链体RNA的自然特征。
