Squaramides and Ureas: A Flexible Approach to Polymerase-Compatible Nucleic Acid Assembly
Squaramides and Ureas: A Flexible Approach to Polymerase-Compatible Nucleic Acid Assembly
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方酰胺和脲：聚合酶兼容核酸组装的灵活方法
DOI：
10.1002/ange.202000209
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发表时间：
2020
影响因子：
--
通讯作者：
Shivalingam A
中科院分区：
文献类型：
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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有力手段。为了恢复这些信息，所产生的连接必须与聚合酶相容。然而，酶促连接是限制性的，并且目前的化学连接方法缺乏灵活性。本文中，描述了基于从最小修饰的3′-和5′-氨基寡核苷酸形成脲和方酰胺人工主链的通用连接平台。一锅连接得到具有优异通读速度的脲键，或在选择性条件下通读的方酰胺键。方酰胺键可以根据需要断裂和重新形成，而稳定的预活化前体寡核苷酸将连接反应的范围扩展到无试剂的温和条件。我们的系统的实用性通过用快速核酸模板依赖性DNA化学连接系统替代RT-qPCR的酶促偏倚RNA至DNA逆转录步骤来证明，该系统允许直接RNA检测。
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.