Cholesterol Anchors Enable Efficient Binding and Intracellular Uptake of DNA Nanostructures.
Cholesterol Anchors Enable Efficient Binding and Intracellular Uptake of DNA Nanostructures.
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DOI：
10.1021/acs.bioconjchem.9b00036
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发表时间：
2019-03
影响因子：
4.7
通讯作者：
William L Whitehouse;J. Noble;M. Ryadnov;S. Howorka
中科院分区：
文献类型：
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作者：
William L Whitehouse;J. Noble;M. Ryadnov;S. Howorka
DNA纳米结构构成了一个快速发展的工具集，用于探索细胞膜功能和细胞内传感或推进生物分子货物进入细胞的递送。与脂质锚定物的化学缀合可以介导DNA纳米结构与合成脂质双层的结合，但这些结构如何与生物膜相互作用并内化细胞尚未显示。在这里，使用原型6-双链体纳米束来研究脂质缀合如何影响DNA细胞结合和内化动力学。使用流式细胞术和共聚焦显微镜评估用一个和三个胆固醇锚修饰的DNA纳米束的细胞相互作用。使用核酸酶消化来区分表面结合的DNA（其是核酸酶可接近的）与内化的DNA。与未修饰的DNA相比，发现三种胆固醇锚定物可增强细胞缔合高达10倍。24 h内细胞被有效内吞。这些结果可以帮助设计受控的DNA结合和运输到细胞中。
DNA nanostructures constitute a rapidly advancing tool-set for exploring cell-membrane functions and intracellular sensing or advancing delivery of biomolecular cargo into cells. Chemical conjugation with lipid anchors can mediate binding of DNA nanostructures to synthetic lipid bilayers, yet how such structures interact with biological membranes and internalize cells has not been shown. Here, an archetypal 6-duplex nanobundle is used to investigate how lipid conjugation influences DNA cell binding and internalization kinetics. Cellular interactions of DNA nanobundles modified with one and three cholesterol anchors were assessed using flow cytometry and confocal microscopy. Nuclease digestion was used to distinguish surface-bound DNA, which is nuclease accessible, from internalized DNA. Three cholesterol anchors were found to enhance cellular association by up to 10-fold when compared with unmodified DNA. The bundles were endocytosed efficiently within 24 h. The results can help design controlled DNA binding and trafficking into cells.