Bailey Jeffrey K, Shen Wen, Liang Xue-Hai, Crooke Stanley T
Department of Core Antisense Research, Ionis Pharmaceuticals, Inc. 2855 Gazelle Court, Carlsbad, CA 92010, USA.
Nucleic Acids Res. 2017 Oct 13;45(18):10649-10671. doi: 10.1093/nar/gkx709.
Antisense oligonucleotides (ASOs) are versatile tools that can regulate multiple steps of RNA biogenesis in cells and living organisms. Significant improvements in delivery, potency, and stability have been achieved through modifications within the oligonucleotide backbone, sugar and heterocycles. However, these modifications can profoundly affect interactions between ASOs and intracellular proteins in ways that are only beginning to be understood. Here, we report that ASOs with specific backbone and sugar modifications can become localized to cytoplasmic ribonucleoprotein granules such as stress granules and those seeded by the aggregation of specific ASO-binding proteins such as FUS/TLS (FUS) and PSF/SFPQ (PSF). Further investigation into the basis for ASO-FUS binding illustrated the importance of ASO backbone and hydrophobic 2' sugar modifications and revealed that the C-terminal region of FUS is sufficient to retain ASOs in cellular foci. Taken together, the results of this study demonstrate that affinities of various nucleic acid binding domains for ASO depend on chemical modifications and further demonstrate how ASO-protein interactions influence the localization of ASOs.
反义寡核苷酸（ASO）是一种多功能工具，可调节细胞和生物体中RNA生物合成的多个步骤。通过对寡核苷酸骨架、糖和杂环进行修饰，在递送、效力和稳定性方面取得了显著改进。然而，这些修饰会以刚刚开始被理解的方式深刻影响ASO与细胞内蛋白质之间的相互作用。在这里，我们报告说，具有特定骨架和糖修饰的ASO可以定位于细胞质核糖核蛋白颗粒，如应激颗粒以及由特定ASO结合蛋白如FUS/TLS（FUS）和PSF/SFPQ（PSF）聚集形成的颗粒。对ASO-FUS结合基础的进一步研究表明了ASO骨架和疏水性2'糖修饰的重要性，并揭示了FUS的C末端区域足以将ASO保留在细胞病灶中。综上所述，本研究结果表明，各种核酸结合结构域对ASO的亲和力取决于化学修饰，并进一步证明了ASO-蛋白质相互作用如何影响ASO的定位。
