CRE recombinase-inducible RNA interference mediated by lentiviral vectors
CRE recombinase-inducible RNA interference mediated by lentiviral vectors
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DOI：
10.1073/pnas.0402107101
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发表时间：
2004-05-11
影响因子：
11.1
通讯作者：
Verma, IM
中科院分区：
文献类型：
--
作者：
Tiscornia, G;Tergaonkar, V;Verma, IM
最近，已经描述了几种设计用于通过使用由聚合酶 III 启动子驱动的小发夹 RNA 来触发 RNA 干扰的系统。在这里，我们报道了一种可由 CRE 重组酶诱导的慢病毒介导的小干扰 RNA 递送系统。该系统由携带小鼠 U6 启动子的慢病毒载体组成，该启动子通过侧翼为修饰的 IoxP 位点的随机 DNA 填充序列与小发夹 RNA 分开。沉默盒只有在通过添加慢病毒载体递送的 CRE 重组酶激活后才会表达。我们使用该系统在体外显示了 GFP 和两个内源基因（肿瘤抑制因子 p53 和 NF-kappaB 转录因子亚基 p65）的特异性下调。此外，p53 和 p65 的下调对下游基因和细胞表型产生了预期的影响。我们预见该系统在体外和体内的多种应用，以组织特异性和局部方式下调特定靶标。
Recently, several systems designed to trigger RNA interference by using small hairpin RNA driven by polymerase III promoters have been described. Here, we report a lentiviral-mediated small interfering RNA delivery system that can be induced by CRE recombinase. The system consists of a lentiviral vector carrying a mouse U6 promoter that is separated from a small hairpin RNA by a random DNA stuffer sequence flanked by modified IoxP sites. The silencing cassette is not expressed until activated by addition of CRE recombinase delivered by a lentiviral vector. We have used this system to show specific down-regulation of GFP and two endogenous genes (the tumor suppressor p53 and the NF-kappaB transcription factor subunit p65) in vitro. Furthermore, down-regulation of both p53 and p65 resulted in the expected effect on downstream genes and cellular phenotype. We foresee multiple applications of this system both in vitro and in vivo to down-regulate specific targets in a tissue-specific and localized manner.