Squaramides and Ureas: A Flexible Approach to Polymerase-Compatible Nucleic Acid Assembly.
Squaramides and Ureas: A Flexible Approach to Polymerase-Compatible Nucleic Acid Assembly.
复制标题
方酰胺和脲：聚合酶兼容核酸组装的灵活方法。
DOI：
10.1002/anie.202000209
复制
发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Shivalingam A
中科院分区：
文献类型：
--
作者：
Shivalingam A
将寡核苷酸连接在一起(连接)是从纳米级检索信息的一种强大手段。为了恢复这一信息，产生的连接必须与聚合酶相容。然而，酶结扎是限制性的，目前的化学结扎方法缺乏灵活性。在此，描述了一种基于最小修饰的3‘-和5’-氨基寡核苷酸形成尿素和方酰胺人工骨架的通用连接平台。一锅法连接提供了具有极佳通读速度的尿素连接，或在选择性条件下通读的方酰胺连接。方酰胺键可以根据需要断开和重整，而稳定的预活化前体寡核苷酸将连接反应的范围扩大到无试剂、温和的条件。我们的系统的实用性是通过用一种快速的核酸模板依赖的DNA化学连接系统取代RT-qPCR中酶偏向的RNA到DNA的反转录步骤来证明的，该系统允许直接检测RNA。
Joining oligonucleotides together (ligation) is a powerful means of retrieving information from the nanoscale. To recover this information, the linkages created must be compatible with polymerases. However, enzymatic ligation is restrictive and current chemical ligation methods lack flexibility. Herein, a versatile ligation platform based on the formation of urea and squaramide artificial backbones from minimally modified 3′‐ and 5′‐amino oligonucleotides is described. One‐pot ligation gives a urea linkage with excellent read‐through speed, or a squaramide linkage that is read‐through under selective conditions. The squaramide linkage can be broken and reformed on demand, while stable pre‐activated precursor oligonucleotides expand the scope of the ligation reaction to reagent‐free, mild conditions. The utility of our system is demonstrated by replacing the enzymatically biased RNA‐to‐DNA reverse transcription step of RT‐qPCR with a rapid nucleic‐acid‐template‐dependent DNA chemical ligation system, that allows direct RNA detection.