Stability of DNA origami nanoarrays in cell lysate.
Stability of DNA origami nanoarrays in cell lysate.
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DOI：
10.1021/nl1040836
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发表时间：
2011-04-13
期刊：
影响因子：
10.8
通讯作者：
Meldrum D
中科院分区：
文献类型：
--
作者：
Mei Q;Wei X;Su F;Liu Y;Youngbull C;Johnson R;Lindsay S;Yan H;Meldrum D
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、确定性蛋白质定位平台和单分子反应观察站。这些应用突出了在活的细胞系统中利用DNA纳米结构的独特性质和生物兼容性的可能性。在这里，我们组装了几种不同形状、大小和探针的DNA折纸纳米结构，并研究了它们与从各种正常和肿瘤细胞系获得的裂解液的相互作用。我们用琼脂糖凝胶电泳法分离和分析了折纸裂解物混合物，并恢复了DNA结构，用于功能分析和后续的显微镜检查。我们的结果表明，DNA折纸纳米结构在细胞裂解液中是稳定的，并且可以很容易地从裂解液混合物中分离出来，而不是天然的单链和双链DNA。原子力显微镜(AFM)和透射电子显微镜(TEM)图像显示，DNA折纸结构在从细胞裂解物中分离并与其目标杂交后完全完整，验证了自组装DNA折纸纳米结构相对于传统寡核苷酸具有更好的结构完整性和功能。细胞裂解液中DNA折纸结构的稳定性和功能性验证了它们在生物学应用中的用途，例如，作为可编程的分子筏或疾病检测平台。
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.
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影响因子：
29.4
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