Chemistry Department, Carnegie Mellon University, 4400 Fifth Avenue Pittsburgh, PA 15213, USA.
Chembiochem. 2011 Jan 3;12(1):125-31. doi: 10.1002/cbic.201000466.
The copper(I)-promoted azide-alkyne cycloaddition reaction (click chemistry) is shown to be compatible with RNA (with free 2'-hydroxyl groups) in spite of the intrinsic lability of RNA. RNA degradation is minimized through stabilization of the Cu(I) in aqueous buffer with acetonitrile as cosolvent and no other ligand; this suggests the general possibility of "ligandless" click chemistry. With the viability of click chemistry validated on synthetic RNA bearing "click"-reactive alkynes, the scope of the reaction is extended to in-vitro-transcribed or, indeed, any RNA, as a click-reactive azide is incorporated enzymatically. Once clickable groups are installed on RNA, they can be rapidly click labeled or conjugated together in click ligations, which may be either templated or nontemplated. In click ligations the resultant unnatural triazole-linked RNA backbone is not detrimental to RNA function, thus suggesting a broad applicability of click chemistry in RNA biological studies.
铜（I）促进的叠氮化物-炔烃环加成反应（点击化学）被证明与 RNA（具有游离的 2'-羟基）兼容，尽管 RNA 具有内在的不稳定性。通过在含有乙腈作为共溶剂的水性缓冲液中稳定 Cu（I）并使用其他配体，可以最小化 RNA 降解；这表明“无配体”点击化学的普遍可能性。通过在带有“点击”反应性炔烃的合成 RNA 上验证点击化学的可行性，该反应的范围扩展到了体外转录的 RNA，或者实际上是任何 RNA，因为可以通过酶促反应引入点击反应性叠氮化物。一旦在 RNA 上安装了可点击的基团，它们可以在点击连接中快速点击标记或连接在一起，这些连接可以是模板化的或非模板化的。在点击连接中，所得的非天然三唑连接的 RNA 骨架对 RNA 功能没有不利影响，因此表明点击化学在 RNA 生物学研究中有广泛的适用性。
