RNase H1-dependent antisense oligonucleotides (ASOs) can degrade complementary RNAs in both the nucleus and the cytoplasm. Since cytoplasmic mRNAs are actively engaged in translation, ASO activity may thus be affected by translating ribosomes that scan the mRNAs. Here we show that mRNAs associated with ribosomes can be cleaved using ASOs and that translation can alter ASO activity. Translation inhibition tends to increase ASO activity when targeting the coding regions of efficiently translated mRNAs, but not nuclear non-coding RNAs or less efficiently translated mRNAs. Increasing the level of RNase H1 protein eliminated the enhancing effects of translation inhibition on ASO activity, suggesting that RNase H1 recruitment to ASO/mRNA heteroduplexes is a rate limiting step and that translating ribosomes can inhibit RNase H1 recruitment. Consistently, ASO activity was not increased by translation inhibition when targeting the 3′ UTRs, independent of the translation efficiency of the mRNAs. Contrarily, the activity of 3′ UTR-targeting ASOs tended to be reduced upon translation inhibition, likely due to decreased accessibility. These results indicate that ASO activity can be affected by the translation process, and the findings also provide important information toward helping better ASO drug design.

中文翻译：

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翻译可影响靶向mRNA的RNase H1依赖性寡核苷酸的反义活性

依赖于RNase H1的反义寡核苷酸（ASO）可以降解细胞核和细胞质中的互补RNA。由于细胞质的mRNA活跃地参与翻译，因此ASO活性可能会受到翻译mRNA的核糖体的翻译的影响。在这里，我们显示可以使用ASO裂解与核糖体相关的mRNA，并且翻译可以改变ASO活性。当靶向有效翻译的mRNA的编码区而不是核非编码RNA或效率较低的翻译的mRNA时，翻译抑制往往会增加ASO活性。RNase H1蛋白水平的增加消除了翻译抑制对ASO活性的增强作用，表明RNase H1募集到ASO / mRNA异源双链体是一个限速步骤，而翻译核糖体可以抑制RNase H1募集。始终如一 靶向3'UTR时，翻译抑制不会提高ASO活性，而与mRNA的翻译效率无关。相反，靶向3′UTR的ASO的活性在翻译抑制时倾向于降低，这可能是由于可及性降低。这些结果表明，ASO活性可能会受到翻译过程的影响，这些发现也为帮助更好地设计ASO药物提供了重要信息。