The synthesis and evaluation of spherical nucleic acids (SNAs) incorporating two physically and chemically distinct classes of oligonucleotides (ODNs) at programmed ratios are described. These SNAs are single entity agents that enter the same target cell at defined stoichiometries, and as such allow one to control important cell signaling and regulatory processes. To study the effect of sequence multiplicity within such structures, we synthesized SNAs consisting of a mixture of class A CpG and class B CpG, immunostimulatory ODNs that activate two different toll-like receptor 9 signaling pathways, each in a sequence-specific fashion. These dual-CpG SNAs exhibit high cellular uptake and codelivery of the two ODNs, relative to mixtures of the linear ODN counterparts, and remain highly associated inside the cell over time. Furthermore, the dual-CpG SNAs augment dendritic cell maturation, compared to the same amounts of oligonucleotides delivered in linear or SNA form but not conjugated to one another. Consequently, these structures constitute a platform for designing oligonucleotide-based combination therapeutics with highly tailorable activities.

中文翻译：

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球形核酸内的序列多重性。

描述了以编程的比例结合了两种物理和化学上不同的寡核苷酸（ODN）的球形核酸（SNA）的合成和评估。这些SNA是单实体代理，它们以定义的化学计量比进入相同的靶细胞，因此可以控制重要的细胞信号传导和调节过程。为了研究此类结构中序列多重性的影响，我们合成了由A类CpG和B类CpG混合物组成的SNA，免疫刺激性ODN，它们激活了两种不同的Toll-like受体9信号通路，各自以序列特异性的方式激活。相对于线性ODN对应物的混合物，这些双CpG SNA表现出较高的细胞摄取和两个ODN的代码传递，并且随着时间的推移在细胞内保持高度关联。此外，与以线性或SNA形式递送但彼此不缀合的相同数量的寡核苷酸相比，双CpG SNA增强了树突状细胞的成熟。因此，这些结构构成了用于设计具有高度可调节活性的基于寡核苷酸的联合治疗药物的平台。