Yamada Ken, Abe Yusuke, Nagatsugi Fumi
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai-shi, Japan.
Curr Protoc Nucleic Acid Chem. 2019 Jun;77(1):e79. doi: 10.1002/cpnc.79. Epub 2019 Mar 12.
This article describes procedures to synthesize 2'-OMe-RNA modified with cross-linkable 2-amino-7-deaza-7-propynyl-6-vinylpurine (ADpVP) and preparation of the RNA-crosslinking experiment in vitro. All synthesis steps yield the desired compound in moderate or high yield without expensive chemical reagents or specific devices. The crosslink-active form of modified RNA can also be purified by commonly used reversed-phase HPLC, can be stored at -80°C after lyophilization for a few days, and is ready to use for crosslinking experiments. This crosslink-active RNA can efficiently form covalent bonds with complementary RNA in a sequence-specific manner. © 2019 by John Wiley & Sons, Inc.
本文描述了合成用可交联的2-氨基-7-脱氮-7-丙炔基-6-乙烯基嘌呤（ADpVP）修饰的2'-O-甲基核糖核酸（2'-OMe-RNA）的方法以及体外核糖核酸交联实验的准备工作。所有合成步骤均能以中等或高产率得到所需化合物，无需昂贵的化学试剂或特殊设备。修饰核糖核酸的交联活性形式也可通过常用的反相高效液相色谱法纯化，冻干后可在-80°C储存数天，并可随时用于交联实验。这种具有交联活性的核糖核酸能够以序列特异性方式与互补核糖核酸高效形成共价键。© 2019约翰威立父子公司版权所有
