Minicircle DNA electrotransfer for efficient tissue-targeted gene delivery
Minicircle DNA electrotransfer for efficient tissue-targeted gene delivery
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DOI：
10.1038/gt.2011.215
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发表时间：
2013-01-01
期刊：
影响因子：
5.1
通讯作者：
Teissie, J.
中科院分区：
文献类型：
--
作者：
Chabot, S.;Orio, J.;Teissie, J.
成功的人类基因治疗或基因疫苗的一个主要问题是安全的高转基因表达水平。基于质粒（非病毒）的物理基因转移方法提供了吸引人的方法，但它们的低效率限制了它们在人体临床前试验中的应用。其中一个限制似乎是质粒的大小，质粒必须通过细胞膜转移到细胞核进行加工。在目前的工作中，我们评估了新一代的DNA载体，以增强基因的转移和表达；微循环，结合电脉冲技术。微环是一种双链环状DNA，由于缺乏细菌序列而缩小了尺寸。结果表明，与全长质粒相比，编码绿色荧光蛋白的电转移小环具有更高的体外转染水平。我们证明了微环高效不是因为细胞毒性降低，而是与细胞更有效的载体摄取有关。载体电转染在体内进行，荧光成像显示，微环电转染可以提高组织靶向基因传递和表达的效率和持续时间。通过结合强大的表达和传递系统，我们为基因治疗和基因疫苗接种的新途径提供了有价值的方法。基因治疗（2013）20,62 -68；doi: 10.1038 / gt.2011.215;2012年1月19日在线发布
A major issue for successful human gene therapy or genetic vaccination is a safe high-transgene expression level. Plasmid-based (non-viral) physical methods of gene transfer offered attracting approaches but their low efficiencies have limited their use in human pre-clinical trials. One of the limits appears to be the size of the plasmid that must be transferred across the cell membrane to the nucleus for its processing. In the present work to enhance gene transfer and expression, we evaluated a new generation of DNA vector; the minicircle, combined with the electropulsation technique. Minicircle is a doubled-stranded circular DNA with reduced size as it is devoid of bacterial sequences. We showed that electrotransferred minicircle encoding green fluorescent protein had higher in vitro transfection level compared with full-length plasmid. We demonstrated that minicircle great efficiency was not because of cellular toxicity decrease but was correlated to more efficient vector uptake by cells. Vector electrotransfection was operated in vivo and, using fluorescence imaging, minicircle electrotransfer was shown to enhance the efficiency and duration of tissue-targeted gene delivery and expression. By combining powerful expression and delivery systems, we have provided a valuable method for new approaches in gene therapy and genetic vaccination. Gene Therapy (2013) 20, 62-68; doi:10.1038/gt.2011.215; published online 19 January 2012