Cre-lox-regulated conditional RNA interference from transgenes
Cre-lox-regulated conditional RNA interference from transgenes
复制标题
DOI：
10.1073/pnas.0403954101
复制
发表时间：
2004-07-13
影响因子：
11.1
通讯作者：
Jacks, T
中科院分区：
文献类型：
--
作者：
Ventura, A;Meissner, A;Jacks, T
我们构建了两种用于条件性、Cre - lox调控的RNA干扰的慢病毒载体。一种载体可实现条件性激活，而另一种允许短发夹RNA（shRNA）表达的条件性失活。前者基于一种策略，即小鼠U6启动子通过包含一个LoxP位点和一个TATA盒之间的杂合体而被修饰。通过在细胞实验中敲低p53、核磷蛋白和DNA甲基转移酶1，证明了使用这些载体有效控制shRNA表达的能力。我们还证明了这种方法在表达Cre的转基因小鼠中实现条件性、组织特异性RNA干扰的有效性。结合不断增加的Cre表达策略，这些载体允许在体内对shRNA表达进行时空控制，并应有助于哺乳动物的功能基因分析。
We have generated two lentiviral vectors for conditional, Cre-loxregulated, RNA interference. One vector allows for conditional activation, whereas the other permits conditional inactivation of short hairpin RNA (shRNA) expression. The former is based on a strategy in which the mouse U6 promoter has been modified by including a hybrid between a LoxP site and a TATA box. The ability to efficiently control shRNA expression by using these vectors was shown in cell-based experiments by knocking down p53, nucleophosmin and DNA methyltransferase 1. We also demonstrate the usefulness of this approach to achieve conditional, tissue-specific RNA interference in Cre-expressing transgenic mice. Combined with the growing array of Cre expression strategies, these vectors allow spatial and temporal control of shRNA expression in vivo and should facilitate functional genetic analysis in mammals.