Self-Assembled Multivalent DNA Nanostructures for Noninvasive Intracellular Delivery of Immunostimulatory CpG Oligonucleotides
Self-Assembled Multivalent DNA Nanostructures for Noninvasive Intracellular Delivery of Immunostimulatory CpG Oligonucleotides
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自组装多价 DNA 纳米结构用于免疫刺激性 CpG 寡核苷酸的无创细胞内递送
DOI：
10.1021/nn202774x
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发表时间：
2011-11-01
期刊：
影响因子：
17.1
通讯作者：
Fan, Chunhai
中科院分区：
文献类型：
--
作者：
Li, Jiang;Pei, Hao;Fan, Chunhai
设计的寡核苷酸可以自组装成具有明确结构和均匀尺寸的DNA纳米结构，这为生物传感，分子成像和药物输送提供了前所未有的机会。在这项工作中，我们已经开发出功能性的，多价的DNA纳米结构，附加非甲基化的CpG基序的三维DNA四面体。这些小尺寸的功能性纳米结构是紧凑的，机械稳定的，并且无细胞毒性。我们已经证明，DNA纳米结构是耐核酸酶降解，并保持基本上完整的胎牛血清和细胞中至少几个小时。值得注意的是，这些功能性纳米结构可以无创和有效地进入巨噬细胞样RAW264.7细胞，而无需转染剂的帮助。在它们被细胞摄取后，CpG基序被Toll样受体9（TLR 9）识别，TLR 9激活下游途径以诱导免疫刺激作用，产生各种促炎细胞因子的高水平分泌，包括肿瘤坏死因子（INF）-α，白细胞介素（IL）-6和IL-12。我们还表明，多价的CpG基序大大增强了纳米结构的免疫刺激作用。鉴于这些功能性纳米结构的高功效和它们的非细胞毒性性质，我们期望DNA纳米结构将成为靶向药物递送的有前途的工具。
Designed oligonucleotides-can self-assemble Into DNA nanostructures with well-defined structures and uniform sizes, which provide unprecedented opportunities for biosensing, molecular imaging, and drug delivery. In this work, we have developed functional, multivalent DNA nanostructures by appending unmethylated CpG motifs to three-dimensional DNA tetrahedra. These small-sized functional nanostructures are compact, mechanically stable, and noncytotoxic. We have demonstrated that DNA nanostructures are resistant to nuclease degradation and remain substantially intact in fetal bovine serum and in cells for at least several hours. Significantly, these functional nanostructures can noninvasively and efficiently enter macrophage-like RAW264.7 cells without the aid of transfection agents. After they are uptaken by cells, CpG motifs are recognized by the Toll-like receptor 9 (TLR9) that activates downstream pathways to induce immunostimulatory effects, producing high-level secretion of various pro-inflammatory cytokines including tumor necrosis factor (INF)-alpha., interleukin (IL)-6, and IL-12. We also show that multivalent CpG motifs greatly enhance the immunostimulatory effect of the nanostructures. Given the high efficacy of these functional nanostructures and their noncytotoxic nature, we expect that DNA nanostructures will become a promising tool for targeted drug delivery.