Off-target effect of endogenous siRNA derived from RMRP in human cells.
Off-target effect of endogenous siRNA derived from RMRP in human cells.
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DOI：
10.3390/ijms14059305
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发表时间：
2013-04-29
影响因子：
5.6
通讯作者：
Masutomi K
中科院分区：
文献类型：
--
作者：
Maida Y;Kyo S;Lassmann T;Hayashizaki Y;Masutomi K
内源性siRNA（endo-siRNA）是植物和蠕虫中RNA沉默的关键调节因子;然而，内源性siRNA在哺乳动物中的生物起源和功能在很大程度上仍然未知。我们以前证明，人端粒酶逆转录酶产生自靶向内源性siRNA从非编码RMRP RNA通过RNA依赖的RNA聚合酶（RdRP）的活性。在这里，我们研究了来自RMRP的endo-siRNA是否靶向RMRP以外的其他基因。四种microRNA靶点预测算法被用于识别endo-siRNA的可能靶点，并且植烷酰辅酶A羟化酶相互作用蛋白样基因（PHYHIPL）被确定为最有希望的候选者。发现PHYHIPL的3′ UTR含有三个可能的靶位点，具有完美的种子配对;这些位点中的每一个的缺失导致上游荧光素酶表达的恢复。此外，序列特异性抑制RMRP衍生的内siRNA增加PHYHIPL mRNA的表达。本文描述的结果表明，endo-siRNA使用的沉默机制与microRNA用于基因沉默的机制相似。据我们所知，该研究首次证实了RdRP活性产生的人内源性siRNA的脱靶效应。
Endogenous siRNAs (endo-siRNAs) are key regulators of RNA silencing in plants and worms; however, the biogenesis and function of endogenous siRNAs in mammals remain largely unknown. We previously demonstrated that human telomerase reverse transcriptase produces a self-targeting endogenous siRNA from non-coding RMRP RNA via RNA-dependent RNA polymerase (RdRP) activity. Here, we investigated whether the endo-siRNA derived from RMRP targets other genes in addition to RMRP. Four algorithms for microRNA target prediction were used to identify possible targets of the endo-siRNA, and the phytanoyl-CoA hydroxylase-interacting protein-like gene (PHYHIPL) was identified as the most promising candidate. The 3′ UTR of PHYHIPL was found to contain three possible target sites with perfect seed pairing; deletion of each of these sites resulted in recovery of upstream luciferase expression. In addition, sequence-specific inhibition of the RMRP-derived endo-siRNA increased expression of PHYHIPL mRNA. The results described here suggest that the endo-siRNA uses silencing mechanisms that are similar to those used by microRNAs for gene silencing. To our knowledge, this study is the first confirmation of the off-target effect of human endogenous siRNA produced by RdRP activity.
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影响因子：
46.9
作者：
Jackson, AL;Bartz, SR;Linsley, PS
通讯作者：
Linsley, PS
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