Stability of DNA origami nanoarrays in cell lysate.

Scaffolded DNA origami, a method to create self-assembled nanostructures with spatially addressable features, has recently been used to develop water-soluble molecular chips for label-free RNA detection, platforms for deterministic protein positioning, and single molecule reaction observatories. These applications highlight the possibility of exploiting the unique properties and biocompatibility of DNA nanostructures in live, cellular systems. Herein, we assembled several DNA origami nanostructures of differing shape, size and probes, and investigated their interaction with lysate obtained from various normal and cancerous cell lines. We separated and analyzed the origami–lysate mixtures using agarose gel electrophoresis and recovered the DNA structures for functional assay and subsequent microscopic examination. Our results demonstrate that DNA origami nanostructures are stable in cell lysate and can be easily separated from lysate mixtures, in contrast to natural, single- and double-stranded DNA. Atomic force microscope (AFM) and transmission electron microscope (TEM) images show that the DNA origami structures are fully intact after separation from cell lysates and hybridize to their targets, verifying the superior structural integrity and functionality of self-assembled DNA origami nanostructures relative to conventional oligonucleotides. The stability and functionality of DNA origami structures in cell lysate validate their use for biological applications, for example, as programmable molecular rafts or disease detection platforms.

支架DNA折纸是一种创建具有空间可寻址特征的自组装纳米结构的方法，最近已被用于开发用于无标记RNA检测的水溶性分子芯片、用于确定性蛋白质定位的平台以及单分子反应观测站。这些应用凸显了在活细胞系统中利用DNA纳米结构的独特性质和生物相容性的可能性。在此，我们组装了几种形状、大小和探针不同的DNA折纸纳米结构，并研究了它们与从各种正常和癌细胞系中获得的裂解物的相互作用。我们使用琼脂糖凝胶电泳分离和分析了折纸 - 裂解物混合物，并回收了DNA结构用于功能测定和随后的显微镜检查。我们的结果表明，与天然的单链和双链DNA相比，DNA折纸纳米结构在细胞裂解物中是稳定的，并且可以很容易地从裂解物混合物中分离出来。原子力显微镜（AFM）和透射电子显微镜（TEM）图像显示，从细胞裂解物中分离后，DNA折纸结构完全完整，并与它们的靶标杂交，验证了自组装DNA折纸纳米结构相对于传统寡核苷酸具有优越的结构完整性和功能性。DNA折纸结构在细胞裂解物中的稳定性和功能性验证了它们在生物应用中的用途，例如，作为可编程分子筏或疾病检测平台。