A regulated, NFκB-assisted import of plasmid DNA into mammalian cell nuclei
A regulated, NFκB-assisted import of plasmid DNA into mammalian cell nuclei
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DOI：
10.1006/mthe.2001.0312
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发表时间：
2001-05-01
影响因子：
12.4
通讯作者：
Reich, Z
中科院分区：
文献类型：
--
作者：
Mesika, A;Grigoreva, I;Reich, Z
通过提高转染效率和对DNA靶向细胞核提供更严格的控制，将增强合成DNA递送系统在人类基因治疗中的成功。在这里，我们使用的DNA载体含有可诱导转录因子NF κ B的重复结合位点，NF κ B通过核输入机制转运到细胞核中。修饰载体的核进入增加了12倍，并与相应的基因表达增加有关。根据它们的位置，结合位点也可以作为转录增强子，使基因表达水平增加19倍。值得注意的是，DNA的核靶向和转基因转录都可以通过调节NF κ B的细胞内分布的外源刺激物来调节。该方法提供了一个框架，用于控制靶向组成型或转录调节的合成载体进入哺乳动物细胞核。
The success of synthetic DNA delivery systems in human gene therapy will be enhanced by increasing transfection efficiencies and by providing tighter control over targeting of the DNA into the nucleus. Here, we used DNA vectors that contain repetitive binding sites for the inducible transcription factor NF kappaB, which is transported into the nucleus by the nuclear import machinery. Nuclear entry of the modified vectors was augmented 12-fold and was associated with corresponding increase in gene expression. Depending on their position, the binding sites could also function as transcriptional enhancers, increasing gene expression levels up to an additional 19-fold. Notably, nuclear targeting of the DNA and transgene transcription could both be regulated by exogenous stimulators which modulate the intracellular distribution of NF kappaB. The approach provides a framework for the controlled targeting of constitutive or transcriptionally regulated synthetic vectors into mammalian cell nuclei.