The SP-C promoter facilitates alveolar type II epithelial cell-specific plasmid nuclear import and gene expression.
The SP-C promoter facilitates alveolar type II epithelial cell-specific plasmid nuclear import and gene expression.
复制标题
作者：
虽然非病毒基因治疗在肺部有很大的应用潜力，但相对缺乏细胞特异性靶向限制了其应用。我们已经开发了一种新的细胞特异性靶向的基础上选择性核输入质粒在非分裂细胞。使用显微注射和原位杂交的方法，我们测试了几个潜在的DNA序列的能力，介导的肺泡II型上皮细胞（ATII）细胞的质粒核输入。其中，只有一个序列内的人表面活性蛋白C（SP-C）启动子能够介导的质粒DNA的核定位，特别是在ATII细胞，但在其他类型的细胞。我们已经将最小输入序列映射到启动子的近端318个核苷酸，并证明NFI、TTF-1和加塔-6的结合位点以及蛋白质本身是输入活性所需的。使用经电穿孔的DNA的气管内递送，我们证明SP-C启动子序列将特异性地增强小鼠肺中ATII细胞中的基因表达。这代表了SP-C启动子的新活性，因此ATII细胞特异性DNA核输入可以证明是ATII细胞中靶向和增强基因表达的安全有效的方法。
Although nonviral gene therapy has great potential for use in the lung, the relative lack of cell-specific targeting has limited its applications. We have developed a new approach for cell-specific targeting based on selective nuclear import of plasmids in non-dividing cells. Using a microinjection and in situ hybridization approach, we tested several potential DNA sequences for the ability to mediate plasmid nuclear import in alveolar type II epithelial (ATII) cells. Of these, only a sequence within the human surfactant protein C (SP-C) promoter was able to mediate nuclear localization of plasmid DNA specifically in ATII cells but not in other cell types. We have mapped the minimal import sequence to the proximal 318 nucleotides of the promoter, and demonstrate that binding sites for NFI, TTF-1, and GATA-6 and the proteins themselves are required for import activity. Using intratracheal delivery of DNA followed by electroporation, we demonstrate that the SP-C promoter sequence will enhance gene expression specifically in ATII cells in mouse lung. This represents a novel activity for the SP-C promoter and thus ATII cell-specific nuclear import of DNA may prove to be a safe and effective method for targeted and enhanced gene expression in ATII cells.
登录
查看更多内容
影响因子：
12.4
作者：
Mesika, A;Grigoreva, I;Reich, Z
通讯作者：
Reich, Z
DOI：
10.1046/j.1432-1327.1999.00796.x
发表时间：
1999-10-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Christophe-Hobertus, C;Duquesne, V;Christophe, D
通讯作者：
Christophe, D
影响因子：
5.1
作者：
Miller, A. M.;Dean, D. A.
通讯作者：
Dean, D. A.
影响因子：
12.4
作者：
Miller, Aaron M.;Munkonge, Felix M.;Dean, David A.
通讯作者：
Dean, David A.
影响因子：
12.4
作者：
Gasiorowski, JZ;Dean, DA
通讯作者：
Dean, DA