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Transfection of small RNAs globally perturbs gene regulation by endogenous microRNAs

热3 gexiaoying 添加于 2010-4-16 20:56 | 1799 次阅读 | 1 个评论

作者
Khan AA, Betel D, Miller ML, Sander C, Leslie CS, Marks DS
摘要
Transfection of small RNAs (such as small interfering RNAs (siRNAs) and microRNAs (miRNAs)) into cells typically lowers expression of many genes. Unexpectedly, increased expression of genes also occurs. We investigated whether this upregulation results from a saturation effect-that is, competition among the transfected small RNAs and the endogenous pool of miRNAs for the intracellular machinery that processes small RNAs. To test this hypothesis, we analyzed genome-wide transcript responses from 151 published transfection experiments in seven different human cell types. We show that targets of endogenous miRNAs are expressed at significantly higher levels after transfection, consistent with impaired effectiveness of endogenous miRNA repression. This effect exhibited concentration and temporal dependence. Notably, the profile of endogenous miRNAs can be largely inferred by correlating miRNA sites with gene expression changes after transfections. The competition and saturation effects have practical implications for miRNA target prediction, the design of siRNA and short hairpin RNA (shRNA) genomic screens and siRNA therapeutics.
详细资料
- 文献种类:期刊
- 期刊名称: Nature Biotechnology
- 期刊缩写: Nat Biotechnol
- 期卷页: 2009年
- ISBN: 1087-0156
标签

miRNA
相关链接 DOI URL

gexiaoying 的文献笔记订阅

转自http://www.xinkexue.com/bbs/viewthread.php?tid=4251 dr.qinan 的笔记

很多做过siRNA/miRNA的人，也许在实验中发现，siRNA/miRNA转染细胞后，目的基因表达不仅没有下调，反而上升！很多人都认为不可思议，更多的时候把这种现象归咎于自己实验操作时的失误，或者甚至放弃进一步探讨。最近的nature biotechnology的一篇paper指出，这种现象是存在的，其原因可能在于细胞内外源性的miRNA与小RNA分子竞争性结合某些细胞内装置，从而引起一种“饱和效应”。这篇文章解决了许多人的困惑，同时也对合理设计siRNA/miRNA不无启示。

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