XRN2 is required for the degradation of target RNAs by RNase H1-dependent antisense oligonucleotides
XRN2 is required for the degradation of target RNAs by RNase H1-dependent antisense oligonucleotides
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DOI：
10.1016/j.bbrc.2015.06.171
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发表时间：
2015-08-21
影响因子：
3.1
通讯作者：
Obika, Satoshi
中科院分区：
文献类型：
--
作者：
Hori, Shin-Ichiro;Yamamoto, Tsuyoshi;Obika, Satoshi
反义寡核苷酸（ASO）可以通过RNA酶H1切割前mRNA和/或成熟mRNA来抑制靶基因的表达。在RNA酶H1的初始核酸内切切割之后，靶RNA被一种尚不清楚的机制降解。为了更好地理解这种降解途径，我们耗尽了两种主要的5'至3'外切核糖核酸酶（XRN）（命名为XRN 1和XRN 2）的表达，并在体外分析了靶RNA的3'片段的水平。我们发现，由阿索产生的靶前体mRNA的3'片段在RNase H1介导的切割后几乎完全被核XRN 2从其5'末端降解，而成熟mRNA的3'片段被XRN 2部分降解。与阿索相比，小干扰RNA（siRNA）仅能降低成熟mRNA的表达水平，3'片段被胞质XRN 1降解。我们的研究结果表明，RNA酶HI依赖性阿索靶向的RNA在细胞核中迅速降解，与siRNA介导的细胞质降解途径相反。(C)2015作者爱思唯尔公司出版
Antisense oligonucleotides (ASOs) can suppress the expression of a target gene by cleaving pre-mRNA and/or mature mRNA via RNase H1. Following the initial endonucleolytic cleavage by RNase H1, the target RNAs are degraded by a mechanism that is poorly understood. To better understand this degradation pathway, we depleted the expression of two major 5' to 3' exoribonucleases (XRNs), named XRN1 and XRN2, and analyzed the levels of 3' fragments of the target RNAs in vitro. We found that the 3' fragments of target pre-mRNA generated by ASO were almost completely degraded from their 5' ends by nuclear XRN2 after RNase H1-mediated cleavage, whereas the 3' fragments of mature mRNA were partially degraded by XRN2. In contrast to ASO, small interference RNA (siRNA) could reduce the expression level of only mature mRNA, and the 3' fragment was degraded by cytoplasmic XRN1. Our findings indicate that the RNAs targeted by RNase HI-dependent ASO are rapidly degraded in the nucleus, contrary to the cytoplasmic degradation pathway mediated by siRNA. (C) 2015 The Authors. Published by Elsevier Inc.