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From the Cover: CRE recombinase-inducible RNA interference mediated by lentiviral vectors

机译:从封面:慢病毒载体介导的CRE重组酶诱导的RNA干扰

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摘要

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 loxP 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-κB 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.
机译:近来,已经描述了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干扰的几种系统。在这里,我们报告了慢病毒介导的小干扰RNA传递系统,可以由CRE重组酶诱导。该系统由携带小鼠U6启动子的慢病毒载体组成,该启动子通过修饰的loxP位点侧翼的随机DNA填充序列与小发夹RNA隔开。沉默盒直到被添加由慢病毒载体递送的CRE重组酶激活才表达。我们已经使用该系统显示了GFP和两个内源基因(肿瘤抑制因子p53和NF-κB转录因子亚基p65)的特异性下调。此外,p53和p65的下调导致对下游基因和细胞表型的预期影响。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化的方式下调特异性靶标。

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