Post-synthesis DNA modifications using a trans-cyclooctene click handle.
Post-synthesis DNA modifications using a trans-cyclooctene click handle.
复制标题
使用反式环辛烯点击手柄进行合成后 DNA 修饰。
DOI：
10.1039/c4ob02031f
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发表时间：
2015
影响因子：
3.2
通讯作者：
Wang,Binghe
中科院分区：
文献类型：
--
作者：
Wang,Ke;Wang,Danzhu;Ji,Kaili;Chen,Weixuan;Zheng,Yueqin;Dai,Chaofeng;Wang,Binghe
DNA合成后修饰是一种非常有用的DNA功能化方法。这是通过使用修饰的NTP来实现的，该修饰的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，手柄可用于PCR后的进一步官能化，优选通过非常快的反应。在这里，我们描述了聚合酶介导的反式环辛烯修饰的胸苷三磷酸（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官能团的四嗪反应。该DNA功能化方法的实用性被证明与硼酸基团和荧光团的掺入。同样的方法也成功地用于修饰用于荧光标记应用的已知适体。
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.