Minicircle DNA vectors devoid of bacterial DNA result in persistent and high-level transgene expression in vivo
Minicircle DNA vectors devoid of bacterial DNA result in persistent and high-level transgene expression in vivo
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DOI：
10.1016/s1525-0016(03)00168-0
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发表时间：
2003-09-01
影响因子：
12.4
通讯作者：
Kay, MA
中科院分区：
文献类型：
--
作者：
Chen, ZY;He, CY;Kay, MA
转基因表达的缺失一直是开发用于治疗人类疾病的非病毒载体的主要障碍。我们以前证明，细菌DNA连接到哺乳动物表达盒导致转基因在体内的转录沉默。为了证实这些研究和开发一种方法来产生一个强大的DNA载体，在体内不沉默，我们开发了一种噬菌体phiC 31整合酶介导的分子内重组技术，以制备小环载体DNA缺乏细菌骨架，然后比较转基因表达谱的小环与不同分子形式的质粒DNA在小鼠中。我们证明，微环DNA缺乏细菌序列表达45倍和560倍以上的血清人因子IX和α 1-抗胰蛋白酶，分别与标准质粒DNA转染到小鼠肝脏。我们的数据表明，微环能够在体内表达高水平和持续水平的治疗产品，并有很大的潜力，作为附加型载体用于治疗各种疾病。
The loss of transgene expression has been a major obstacle to the development of nonviral vectors for the treatment of human diseases. We previously demonstrated that bacterial DNA linked to a mammalian expression cassette resulted in transcriptional silencing of the transgene in vivo. To confirm these studies and develop a means to produce a robust DNA vector that is not silenced in vivo, we developed a phage phiC31 integrase-mediated intramolecular recombination technology to prepare minicircle vector DNA devoid of the bacterial backbone and then compared the transgene expression profile of the minicircle with different molecular forms of plasmid DNAs in mice. We demonstrate that minicircular DNAs devoid of bacterial sequences expressed 45- and 560-fold more serum human factor IX and alpha1-antitrypsin, respectively, compared to standard plasmid DNAs transfected into mouse liver. Our data suggest that minicircles are capable of expressing high and persistent levels of therapeutic products in vivo and have a great potential to serve as episomal vectors for the treatment of a wide variety of diseases.