Enhancing the stability of DNA origami nanostructures: staple strand redesign versus enzymatic ligation
Enhancing the stability of DNA origami nanostructures: staple strand redesign versus enzymatic ligation
复制标题
DOI：
10.1039/c9nr04460d
复制
发表时间：
2019-09-21
期刊：
影响因子：
6.7
通讯作者：
Krainer, Georg
中科院分区：
文献类型：
--
作者：
Ramakrishnan, Saminathan;Schaerfen, Leonard;Krainer, Georg
DNA 折纸结构已发展成为分子科学和纳米技术领域的多功能工具。然而，目前许多潜在的应用因其稳定性差而受到阻碍，特别是在变性条件下。在这里，我们提出并评估了两种增强 DNA 折纸稳定性的简单方法。在第一种方法中，我们通过将寡核苷酸与相邻序列合并来提高九个关键主链的解链温度。在第二种方法中，我们通过直接酶促连接所有可接近的短链末端来提高整体稳定性。通过使用原子力显微镜监测通过这些方法修改的原型三角形DNA折纸的逐渐尿素诱导变性，我们表明，对一些关键主链进行合理的重新设计可以在高变性剂浓度和高温下显着提高整体稳定性。此外，酶连接产生的 DNA 纳米结构在高达 37 摄氏度和 6 M 尿素存在下具有优异的稳定性，而不会损害其形状。当需要在恶劣条件下保持结构完整性时，这种生物正交方法很容易适用于其他 DNA 折纸结构，无需进行合成核苷酸修饰。
DNA origami structures have developed into versatile tools in molecular sciences and nanotechnology. Currently, however, many potential applications are hindered by their poor stability, especially under denaturing conditions. Here we present and evaluate two simple approaches to enhance DNA origami stability. In the first approach, we elevated the melting temperature of nine critical staple strands by merging the oligonucleotides with adjacent sequences. In the second approach, we increased the global stability by enzymatically ligating all accessible staple strand ends directly. By monitoring the gradual urea-induced denaturation of a prototype triangular DNA origami modified by these approaches using atomic force microscopy, we show that rational redesign of a few, critical staple strands leads to a considerable increase in overall stability at high denaturant concentration and elevated temperatures. In addition, enzymatic ligation yields DNA nanostructures with superior stability at up to 37 degrees C and in the presence of 6 M urea without impairing their shape. This bio-orthogonal approach is readily adaptable to other DNA origami structures without the need for synthetic nucleotide modifications when structural integrity under harsh conditions is required.