Stable knockdown of microRNA in vivo by lentiviral vectors
Stable knockdown of microRNA in vivo by lentiviral vectors
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DOI：
10.1038/nmeth.1277
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发表时间：
2009-01-01
期刊：
影响因子：
48
通讯作者：
Naldini, Luigi
中科院分区：
文献类型：
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作者：
Gentner, Bernhard;Schira, Giulia;Naldini, Luigi
在体内研究微小RNA（microRNA）的功能需要遗传学策略来产生功能缺失表型。我们利用慢病毒载体，通过从聚合酶II启动子过表达微小RNA的靶序列，稳定且特异性地敲低微小RNA。这些载体有效地抑制了报告基因构建体以及天然微小RNA靶标的调控。我们利用稳定过表达miR - 223靶序列的造血干细胞进行骨髓重建，以模拟miR - 223基因敲除小鼠的表型，这表明在体内对微小RNA功能产生了强烈的干扰。
Studying microRNA function in vivo requires genetic strategies to generate loss-of-function phenotypes. We used lentiviral vectors to stably and specifically knock down microRNA by overexpressing microRNA target sequences from polymerase II promoters. These vectors effectively inhibited regulation of reporter constructs and natural microRNA targets. We used bone marrow reconstitution with hematopoietic stem cells stably overexpressing miR-223 target sequence to phenocopy the genetic miR-223 knockout mouse, indicating robust interference of microRNA function in vivo.