Gene silencing by the tRNA maturase tRNase ZL under the direction of small-guide RNA
Gene silencing by the tRNA maturase tRNase ZL under the direction of small-guide RNA
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DOI：
10.1038/sj.gt.3302841
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发表时间：
2007-01-01
期刊：
影响因子：
5.1
通讯作者：
Nashimoto, M.
中科院分区：
文献类型：
--
作者：
Nakashima, A.;Takaku, H.;Nashimoto, M.
我们一直在开发一种独特的系统，用于通过在小向导RNA（sgRNA）的指导下由长形式的tRNA 3 '加工核糖核酸内切酶（tRNase Z（L））切割特定mRNA来下调基因表达。然而，该系统的功效和tRNase Z（L）在活细胞中的参与尚不清楚。在这里，我们表明，通过靶向外源荧光素酶基因，sgRNA/tRNase Z（L）方法的功效可以变得与RNA干扰技术的功效相当，并且基因沉默是由于由sgRNA指导的tRNase Z（L）而不是由于简单的反义效应。我们还表明，tRNase Z（L）与sgRNA一起可以通过降解细胞培养物中的内源性人类基因Bcl-2和糖原合成酶激酶-3 β的mRNA来下调它们的表达。此外，我们证明了出生后小鼠肝脏中的基因表达可以被仅7个核苷酸的sgRNA抑制。这些数据表明，sgRNA可能被用作治疗剂来治疗疾病，如癌症和艾滋病。
We have been developing a unique system for the down-regulation of a gene expression through cutting a specific mRNA by the long form of tRNA 3'-processing endoribonuclease (tRNase Z(L)) under the direction of small-guide RNA (sgRNA). However, the efficacy of this system and the involvement of tRNase Z(L) in the living cells were not clear. Here we show, by targeting the exogenous luciferase gene, that the efficacy of the sgRNA/tRNase Z(L) method can become comparable to that of the RNA interference technology and that the gene silencing is owing to tRNase Z(L) directed by sgRNA not owing to a simple antisense effect. We also show that tRNase Z(L) together with sgRNA can downregulate expression of the endogenous human genes Bcl-2 and glycogen synthase kinase-3 beta by degrading their mRNAs in cell culture. Furthermore, we demonstrate that a gene expression in the livers of postnatal mice can be inhibited by an only seven-nucleotide sgRNA. These data suggest that sgRNA might be utilized as therapeutic agents to treat diseases such as cancers and AIDS.