Department of Chemistry, Stanford University, Stanford, CA, 94305, USA.
Angew Chem Int Ed Engl. 2018 Mar 12;57(12):3059-3063. doi: 10.1002/anie.201708696. Epub 2018 Feb 22.
We describe a selective and mild chemical approach for controlling RNA hybridization, folding, and enzyme interactions. Reaction of RNAs in aqueous buffer with an azide-substituted acylating agent (100-200 mm) yields several 2'-OH acylations per RNA strand in as little as 10 min. This poly-acylated ("cloaked") RNA is strongly blocked from hybridization with complementary nucleic acids, from cleavage by RNA-processing enzymes, and from folding into active aptamer structures. Importantly, treatment with a water-soluble phosphine triggers a Staudinger reduction of the azide groups, resulting in spontaneous loss of acyl groups ("uncloaking"). This fully restores RNA folding and biochemical activity.
我们描述了一种选择性的和温和的化学方法来控制 RNA 的杂交、折叠和酶相互作用。在水溶液缓冲液中，用取代的叠氮酰化试剂反应（100-200 mM），在短短 10 分钟内每个 RNA 链上可产生几个 2'-OH 酰化反应。这种多酰化的（“伪装”）RNA 强烈地阻止与互补核酸杂交，阻止 RNA 加工酶切割，并阻止折叠成有活性的适体结构。重要的是，用水溶性膦处理会引发叠氮化物的斯塔蒂格还原，导致酰基的自发丢失（“解伪装”）。这完全恢复了 RNA 的折叠和生化活性。
