使用2'-甲氧基乙基进行的位点特异性修饰可提高siRNA的特异性和活性。,Molecular Therapy - Nucleic Acids

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使用2'-甲氧基乙基进行的位点特异性修饰可提高siRNA的特异性和活性。
Molecular Therapy - Nucleic Acids ( IF 6.5 ) Pub Date : 2017-12-17 , DOI: 10.1016/j.omtn.2017.10.003
Xinyun Song ₁ , Xiaoxia Wang ₁ , Yuan Ma ₂ , Zicai Liang ₁ , Zhenjun Yang ₂ , Huiqing Cao ₁

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在针对人类疾病的基于小干扰RNA（siRNA）的疗法方面已取得了快速进展，但是合理优化siRNA的高特异性和有效沉默仍然是一个挑战。在这项研究中，我们探讨了在siRNA切割位点进行化学修饰的作用。我们发现在位置9和10的修饰显着降低了未修饰的siRNA链的沉默能力，但被修饰的链具有很好的耐受性。有趣的是，在裂解位点添加2'-甲氧基乙基（MOE）基团可通过促进修饰链的定向RNA诱导的沉默复合物（RISC）加载，提高siRNA的特异性和沉默活性。此外，我们将一条链的9和10位的MOE修饰与2' -O-甲基化（OMe）在另一条链的14位，并发现了一种协同效应，可提高siRNA的特异性。使用靶向siRNA的内源基因细胞间粘附分子1（ICAM1）验证了组合修饰的令人惊讶的有益效果。我们发现组合的修改消除了其脱靶效应。总之，我们建立了使用位点特异性MOE修饰优化siRNA的有效策略。该发现可以允许就活性和特异性而言创建用于治疗的优良siRNA。我们发现组合的修改消除了其脱靶效应。总之，我们建立了使用位点特异性MOE修饰优化siRNA的有效策略。该发现可以允许就活性和特异性而言创建用于治疗的优良siRNA。我们发现组合的修改消除了其脱靶效应。总之，我们建立了使用位点特异性MOE修饰优化siRNA的有效策略。该发现可以允许就活性和特异性而言创建用于治疗的优良siRNA。

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Site-Specific Modification Using the 2'-Methoxyethyl Group Improves the Specificity and Activity of siRNAs.

Rapid progress has been made toward small interfering RNA (siRNA)-based therapy for human disorders, but rationally optimizing siRNAs for high specificity and potent silencing remains a challenge. In this study, we explored the effect of chemical modification at the cleavage site of siRNAs. We found that modifications at positions 9 and 10 markedly reduced the silencing potency of the unmodified strand of siRNAs but were well tolerated by the modified strand. Intriguingly, addition of the 2'-methoxyethyl (MOE) group at the cleavage site improved both the specificity and silencing activity of siRNAs by facilitating the oriented RNA-induced silencing complex (RISC) loading of the modified strand. Furthermore, we combined MOE modifications at positions 9 and 10 of one strand together with 2'-O-methylation (OMe) at position 14 of the other strand and found a synergistic effect that improved the specificity of siRNAs. The surprisingly beneficial effect of the combined modification was validated using siRNA-targeting endogenous gene intercellular adhesion molecule 1 (ICAM1). We found that the combined modifications eliminated its off-target effects. In conclusion, we established effective strategies to optimize siRNAs using site-specific MOE modifications. The findings may allow the creation of superior siRNAs for therapy in terms of activity and specificity.

更新日期：2019-11-01

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