Ahmadi Yasaman, Barisic Ivan
Molecular Diagnostics, Centre for Health and Bioresources, AIT Austrian Institute of Technology GmbH;
J Vis Exp. 2019 Jan 19(143). doi: 10.3791/58771.
DNA origami nanostructures hold an immense potential to be used for biological and medical applications. However, low-salt conditions and nucleases in physiological fluids induce denaturation and degradation of self-assembled DNA nanostructures. In non-viral gene delivery, enzymatic degradation of DNA is overcome by the encapsulation of the negatively charged DNA in a cationic shell. Herein, inspired by gene delivery advancements, a simple, one-step and robust methodology is presented for the stabilization of DNA origami nanostructures by coating them with chitosan and linear polyethyleneimine. The polycation coating efficiently protects DNA origami nanostructures in Mg-depleted and nuclease-rich media. This method also preserves the full addressability of enzyme- and aptamer-based functionalization of DNA nanostructures.
DNA折纸纳米结构在生物和医学应用方面具有巨大的应用潜力。然而,生理流体中的低盐条件和核酸酶会导致自组装DNA纳米结构变性和降解。在非病毒基因递送中,通过将带负电荷的DNA包裹在阳离子壳中来克服DNA的酶促降解。在此,受基因递送进展的启发,提出了一种简单、一步且稳健的方法,通过用壳聚糖和线性聚乙烯亚胺包被来稳定DNA折纸纳米结构。聚阳离子涂层在缺镁和富含核酸酶的介质中能有效保护DNA折纸纳米结构。该方法还保留了基于酶和适体的DNA纳米结构功能化的完全可寻址性。