mRNA levels can be reduced by antisense oligonucleotides via no-go decay pathway
mRNA levels can be reduced by antisense oligonucleotides via no-go decay pathway
复制标题
DOI：
10.1093/nar/gkz500
复制
发表时间：
2019-07-26
影响因子：
14.9
通讯作者：
Crooke, Stanley T.
中科院分区：
文献类型：
--
作者：
Liang, Xue-hai;Nichols, Joshua G.;Crooke, Stanley T.
反义技术可以通过RNaseH1或RISC途径降低基因表达，通过调控剪接或翻译提高基因表达。在这里，我们证明了反义寡核苷酸(ASO)可以通过作用于NO-GO衰变途径来降低mRNA水平。硫代硫酸盐ASO被2‘-O-甲氧基乙基完全修饰后，以翻译依赖、RNaseH1不依赖的方式靶向mRNAs编码区时，mRNA水平降低。激活这一衰变途径的ASO在编码区的3‘端附近杂交。虽然一些ASO诱导了无意义介导的衰变，但另一些ASO通过NO-GO衰变途径降低了mRNA水平，因为NO-GO衰变途径所需的蛋白质Pelo/HBS1L的缺失降低了这些ASO的活性。ASO长度和化学修饰影响这些试剂的效果。在不同的转录本和不同的细胞系中观察到了这种非缺口ASO诱导的mRNA减少。因此，我们的研究确定了一种使用ASOS降解mRNAs的新机制，增加了一种新的反义方法来调节基因表达。这也有助于解释为什么一些完全修饰的ASO导致RNA靶标减少，尽管它们不能作为RNaseH1的底物。
Antisense technology can reduce gene expression via the RNase H1 or RISC pathways and can increase gene expression through modulation of splicing or translation. Here, we demonstrate that antisense oligonucleotides (ASOs) can reduce mRNA levels by acting through the no-go decay pathway. Phosphorothioate ASOs fully modified with 2'-O-methoxyethyl decreased mRNA levels when targeted to coding regions of mRNAs in a translation-dependent, RNase H1-independent manner. The ASOs that activated this decay pathway hybridized near the 3' end of the coding regions. Although some ASOs induced nonsense-mediated decay, others reduced mRNA levels through the no-go decay pathway, since depletion of PELO/HBS1L, proteins required for no-go decay pathway activity, decreased the activities of these ASOs. ASO length and chemical modification influenced the efficacy of these reagents. This non-gapmer ASO-induced mRNA reduction was observed for different transcripts and in different cell lines. Thus, our study identifies a new mechanism by which mRNAs can be degraded using ASOs, adding a new antisense approach to modulation of gene expression. It also helps explain why some fully modified ASOs cause RNA target to be reduced despite being unable to serve as substrates for RNase H1.