Antisense oligonucleotides (ASOs) regulate gene expression by binding to complementary target RNA, and ASOs can be designed to take advantage of a growing array of post RNA binding molecular mechanisms. Intracellular trafficking of ASOs influences their efficacy. We have identified a number of membrane-less structures in the nucleus, nucleolus, and cytoplasm where phosphorothioate-modified ASOs (PS-ASOs) accumulate and have shown that PS-ASOs can induce the formation of new nuclear structures such as PS-bodies and paraspeckle-like structures. In this study, we report that PS-ASOs can localize to cytoplasmic processing bodies (P-bodies) and increase the number of P-bodies in cells. The antisense activity of PS-ASOs was not affected by the absence of essential P-body assembly proteins DDX6 and LSm14A. Moreover, the effects of PS-ASOs on P-body assembly were independent of their antisense activities. The phosphorothioate modification stabilizes the association between ASOs and cellular proteins and is essential for the P-body localization of ASOs. Since PS-ASOs bind to major P-body components, PS-ASOs may serve as scaffolds for P-body formation. Taken together, these results indicate that interactions of PS-ASO with proteins, rather than antisense activities, are essential for the dynamic interplay between PS-ASOs and P-bodies.

中文翻译：

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硫代磷酸反义寡核苷酸结合P体蛋白并介导P体组装。

反义寡核苷酸（ASO）通过与互补靶RNA结合来调节基因表达，并且可以将ASO设计为利用不断增长的后RNA结合分子机制。细胞内ASO的运输会影响其功效。我们已经确定了硫代磷酸酯修饰的ASO（PS-ASO）积累的核，核仁和细胞质中的许多无膜结构，并表明PS-ASO可以诱导新的核结构形成，例如PS体和散斑状结构。在这项研究中，我们报道PS-ASOs可以定位于细胞质加工体（P体）并增加细胞中P体的数量。PS-ASO的反义活性不受必需的P体装配蛋白DDX6和LSm14A的影响。此外，PS-ASO对P体组装的影响与其反义活性无关。硫代磷酸酯修饰可稳定ASO与细胞蛋白之间的缔合，对于ASO的P体定位至关重要。由于PS-ASO与主要的P体成分结合，因此PS-ASO可以充当P体形成的支架。两者合计，这些结果表明PS-ASO与蛋白质的相互作用，而不是反义活性，对于PS-ASO与P体之间的动态相互作用至关重要。