State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, No. 38 Xueyuan Road, Beijing 100191, China.
ACS Appl Mater Interfaces. 2020 Mar 18;12(11):12584-12590. doi: 10.1021/acsami.0c00923. Epub 2020 Mar 9.
Circular oligodeoxynucleotides (c-ODNs) have their particular characteristics in topological properties. However, different from oligoribonucleotides, enzymatic synthesis of small c-ODNs is still challenging using conventional methods. Herein, we successfully achieved highly efficient cyclization of linear single-stranded ODNs using T4 DNA ligase simply through the frozen/lyophilization/cyclization (FLC) method. We successfully shortened the cyclization length of linear ODNs to 20 nt (l-ODN 20) with up to 63% yield, which was never achieved before through normal enzymatic methods. With the efficient FLC method, we further developed "DNA interlocks" which were intercross-linked with multiple c-ODNs using the one-pot FLC method. This FLC strategy provides a powerful, cheap, and convenient method to synthesize small c-ODNs for studying DNA nanotechnology and paves the way to achieve future deciphering of c-ODN functions.
环状寡脱氧核苷酸（c-ODNs）在拓扑性质上具有其独特的特征。然而，与寡核糖核苷酸不同，使用常规方法对小的 c-ODNs 进行酶促合成仍然具有挑战性。在此，我们通过简单的 T4 DNA 连接酶冷冻/冻干/环化（FLC）方法，成功地实现了线性单链 ODN 的高效环化。我们成功地将线性 ODN 的环化长度缩短至 20 个核苷酸（l-ODN 20），产率高达 63%，这是以前通过常规酶促方法从未实现的。通过高效的 FLC 方法，我们进一步开发了“DNA 互锁”，它可以通过一锅 FLC 方法与多个 c-ODNs 交叉连接。这种 FLC 策略为研究 DNA 纳米技术提供了一种强大、廉价和方便的合成小 c-ODNs 的方法，并为实现未来对 c-ODN 功能的破译铺平了道路。
