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Cryopreservation of DNA Origami Nanostructures
Small ( IF 13.0 ) Pub Date : 2020-03-04 , DOI: 10.1002/smll.201905959
Yang Xin 1 , Charlotte Kielar 1 , Siqi Zhu 1 , Christoph Sikeler 2 , Xiaodan Xu 1 , Christin Möser 3, 4 , Guido Grundmeier 1 , Tim Liedl 2 , Amelie Heuer‐Jungemann 2 , David M. Smith 3, 5 , Adrian Keller 1
Affiliation  

Although DNA origami nanostructures have found their way into numerous fields of fundamental and applied research, they often suffer from rather limited stability when subjected to environments that differ from the employed assembly conditions, that is, suspended in Mg2+‐containing buffer at moderate temperatures. Here, means for efficient cryopreservation of 2D and 3D DNA origami nanostructures and, in particular, the effect of repeated freezing and thawing cycles are investigated. It is found that, while the 2D DNA origami nanostructures maintain their structural integrity over at least 32 freeze–thaw cycles, ice crystal formation makes the DNA origami gradually more sensitive toward harsh sample treatment conditions. Whereas no freeze damage could be detected in 3D DNA origami nanostructures subjected to 32 freeze–thaw cycles, 1000 freeze–thaw cycles result in significant fragmentation. The cryoprotectants glycerol and trehalose are found to efficiently protect the DNA origami nanostructures against freeze damage at concentrations between 0.2 × 10−3 and 200 × 10−3 m and without any negative effects on DNA origami shape. This work thus provides a basis for the long‐term storage of DNA origami nanostructures, which is an important prerequisite for various technological and medical applications.

中文翻译:

DNA折纸纳米结构的冷冻保存

尽管DNA折纸纳米结构已进入基础研究和应用研究的众多领域,但在不同于所用装配条件的环境(即悬浮在Mg 2+中)下,它们经常遭受相当有限的稳定性。含适量温度的缓冲液。在此,研究了有效冷冻保存2D和3D DNA折纸纳米结构的方法,尤其是研究了反复冷冻和解冻循环的效果。发现,虽然二维DNA折纸纳米结构在至少32个冻融循环中保持其结构完整性,但冰晶的形成使DNA折纸对苛刻的样品处理条件逐渐变得更加敏感。尽管经过32次冻融循环的3D DNA折纸纳米结构中没有发现冻损,但1000次冻融循环却导致了明显的碎片。发现冷冻保护剂甘油和海藻糖可以有效保护DNA折纸纳米结构,使其在0.2×10 -3到200×10的浓度下免受冻害。-3 m,对DNA折纸形状没有任何负面影响。因此,这项工作为DNA折纸纳米结构的长期存储提供了基础,这是各种技术和医学应用的重要前提。
更新日期:2020-04-03
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