The RNA Institute, University at Albany, State University of New York, Albany, NY, 12222, USA.
Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
Small. 2023 Sep;19(39):e2300040. doi: 10.1002/smll.202300040. Epub 2023 Jun 1.
The programmable nature of DNA allows the construction of custom-designed static and dynamic nanostructures, and assembly conditions typically require high concentrations of magnesium ions that restricts their applications. In other solution conditions tested for DNA nanostructure assembly, only a limited set of divalent and monovalent ions are used so far (typically Mg and Na ). Here, we investigate the assembly of DNA nanostructures in a wide variety of ions using nanostructures of different sizes: a double-crossover motif (76 bp), a three-point-star motif (~134 bp), a DNA tetrahedron (534 bp) and a DNA origami triangle (7221 bp). We show successful assembly of a majority of these structures in Ca , Ba , Na , K and Li and provide quantified assembly yields using gel electrophoresis and visual confirmation of a DNA origami triangle using atomic force microscopy. We further show that structures assembled in monovalent ions (Na , K and Li ) exhibit up to a 10-fold higher nuclease resistance compared to those assembled in divalent ions (Mg , Ca and Ba ). Our work presents new assembly conditions for a wide range of DNA nanostructures with enhanced biostability.
DNA 的可编程性允许构建定制设计的静态和动态纳米结构,而组装条件通常需要高浓度的镁离子,这限制了它们的应用。在迄今为止测试的用于 DNA 纳米结构组装的其他溶液条件下,只使用了有限的一组二价和一价离子(通常是 Mg 和 Na)。在这里,我们使用不同尺寸的纳米结构研究了在各种离子中组装 DNA 纳米结构的情况:双交叉基元(76 bp)、三点星基元(~134 bp)、DNA 四面体(534 bp)和 DNA 折纸三角形(7221 bp)。我们展示了这些结构中的大多数在 Ca、Ba、Na、K 和 Li 中成功组装,并使用凝胶电泳进行了定量组装产率的测量,并用原子力显微镜对 DNA 折纸三角形进行了可视化确认。我们进一步表明,与在二价离子(Mg、Ca 和 Ba)中组装的结构相比,在单价离子(Na、K 和 Li)中组装的结构具有高达 10 倍的更高的核酸酶抗性。我们的工作为具有增强的生物稳定性的广泛的 DNA 纳米结构提供了新的组装条件。