使用反式环辛烯点击手柄进行合成后DNA修饰。
Post-synthesis DNA modifications using a trans-cyclooctene click handle.
作者信息
Wang Ke, Wang Danzhu, Ji Kaili, Chen Weixuan, Zheng Yueqin, Dai Chaofeng, Wang Binghe
机构信息
Department of Chemistry, Center for Diagnostics and Therapeutics, Georgia State University, Atlanta, GA 30302-3965, USA.
出版信息
Org Biomol Chem. 2015 Jan 21;13(3):909-15. doi: 10.1039/c4ob02031f.
Abstract
Post-synthesis DNA modification is a very useful method for DNA functionalization. This is achieved by using a modified NTP, which has a handle for further modifications, replacing the corresponding natural NTP in polymerase-catalyzed DNA synthesis. Subsequently, the handle can be used for further functionalization after PCR, preferably through a very fast reaction. Herein we describe polymerase-mediated incorporation of trans-cyclooctene modified thymidine triphosphate (TCO-TTP). Subsequently, the trans-cyclooctene group was reacted with a tetrazine tethered to other functional groups through a very fast click reaction. The utility of this DNA functionalization method was demonstrated with the incorporation of a boronic acid group and a fluorophore. The same approach was also successfully used in modifying a known aptamer for fluorescent labelling applications.
摘要
合成后DNA修饰是一种非常有用的DNA功能化方法。这是通过使用一种修饰的NTP来实现的，该NTP具有用于进一步修饰的手柄，在聚合酶催化的DNA合成中取代相应的天然NTP。随后，该手柄可在PCR后用于进一步功能化，最好通过非常快速的反应。在此，我们描述了聚合酶介导的反式环辛烯修饰的三磷酸胸苷（TCO-TTP）的掺入。随后，反式环辛烯基团通过非常快速的点击反应与连接到其他功能基团的四嗪反应。通过硼酸基团和荧光团的掺入证明了这种DNA功能化方法的实用性。相同的方法也成功用于修饰已知适体以用于荧光标记应用。
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