A functional assay for microRNA target identification and validation.
A functional assay for microRNA target identification and validation.
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DOI：
10.1093/nar/gks145
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发表时间：
2012-05
影响因子：
14.9
通讯作者：
Mufti GJ
中科院分区：
文献类型：
--
作者：
Gäken J;Mohamedali AM;Jiang J;Malik F;Stangl D;Smith AE;Chronis C;Kulasekararaj AG;Thomas NS;Farzaneh F;Tavassoli M;Mufti GJ
MicroRNA（miRNA）是一类小RNA分子，其调节许多关键的细胞过程并与部分互补序列结合，从而导致其靶基因的下调。由于miRNA与其靶位点的不完全同源性，miRNA靶基因的鉴定是困难的，并且目前基于预测给定miRNA的大量潜在靶标的计算算法。为了能够鉴定生物学相关的miRNA靶点，我们描述了一种基于3′-UTR富集文库和正/负选择策略的新型功能测定。作为原理证明，我们使用mir-130 a及其经验证的靶MAFB来测试该策略。通过蛋白质印迹分析和敲除实验鉴定MAFB和五个另外的靶标并随后确认其为mir-130 a靶标，验证了用于miRNA靶标的功能鉴定的该策略。
MicroRNAs (miRNA) are a class of small RNA molecules that regulate numerous critical cellular processes and bind to partially complementary sequences resulting in down-regulation of their target genes. Due to the incomplete homology of the miRNA to its target site identification of miRNA target genes is difficult and currently based on computational algorithms predicting large numbers of potential targets for a given miRNA. To enable the identification of biologically relevant miRNA targets, we describe a novel functional assay based on a 3′-UTR-enriched library and a positive/negative selection strategy. As proof of principle we have used mir-130a and its validated target MAFB to test this strategy. Identification of MAFB and five additional targets and their subsequent confirmation as mir-130a targets by western blot analysis and knockdown experiments validates this strategy for the functional identification of miRNA targets.
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影响因子：
4.1
作者：
Beitzinger, Michaela;Peters, Lasse;Meister, Gunter
通讯作者：
Meister, Gunter
影响因子：
4.5
作者：
Easow, George;Teleman, Aurelio A.;Cohen, Stephen M.
通讯作者：
Cohen, Stephen M.
影响因子：
4.7
作者：
Sorrentino, Antonio;Liu, Chang-Gong;Ferlini, Cristiano
通讯作者：
Ferlini, Cristiano
影响因子：
16
作者：
Lal A;Navarro F;Maher CA;Maliszewski LE;Yan N;O'Day E;Chowdhury D;Dykxhoorn DM;Tsai P;Hofmann O;Becker KG;Gorospe M;Hide W;Lieberman J
通讯作者：
Lieberman J
影响因子：
20.3
作者：
Chen, Yun;Gorski, David H.
通讯作者：
Gorski, David H.