Translation efficiency of mRNAs is increased by antisense oligonucleotides targeting upstream open reading frames
Translation efficiency of mRNAs is increased by antisense oligonucleotides targeting upstream open reading frames
复制标题
DOI：
10.1038/nbt.3589
复制
发表时间：
2016-08-01
影响因子：
46.9
通讯作者：
Crooke, Stanley T.
中科院分区：
文献类型：
--
作者：
Liang, Xue-hai;Shen, Wen;Crooke, Stanley T.
增加体内治疗性蛋白质的水平仍然具有挑战性。反义寡核苷酸（ASO）通常用于下调基因表达（1）或修饰RNA剪接（2，3），但反义技术以前尚未用于直接增加选定蛋白质的产量。在这里，我们使用了一类修饰的ASO，其结合上游开放阅读框（uORF）中的mRNA序列，以特异性地增加从下游初级ORF（pORF）翻译的蛋白质的量。使用阿索处理，我们在人和小鼠细胞中以剂量依赖性方式将从四个基因表达的蛋白质的量增加了30-150%。值得注意的是，用阿索全身治疗小鼠导致LRPPRC的蛋白质增加接近80%。ASO介导的蛋白质表达增加是序列特异性的，发生在翻译水平，并依赖于解旋酶活性。我们还发现RNA修饰的类型和修饰核苷酸在ASO中的位置影响pORF的翻译。ASO是一类具有广泛用途的有用治疗剂。
Increasing the levels of therapeutic proteins in vivo remains challenging. Antisense oligonucleotides (ASOs) are often used to downregulate gene expression(1) or to modify RNA splicing(2,3,) but antisense technology has not previously been used to directly increase the production of selected proteins. Here we used a class of modified ASOs that bind to mRNA sequences in upstream open reading frames (uORFs) to specifically increase the amounts of protein translated from a downstream primary ORF (pORF). Using ASO treatment, we increased the amount of proteins expressed from four genes by 30-150% in a dose-dependent manner in both human and mouse cells. Notably, systemic treatment of mice with ASO resulted in an similar to 80% protein increase of LRPPRC. The ASO-mediated increase in protein expression was sequence-specific, occurred at the level of translation and was dependent on helicase activity. We also found that the type of RNA modification and the position of modified nucleotides in ASOs affected translation of a pORF. ASOs are a useful class of therapeutic agents with broad utility.