Plasmid DNA electrotransfer for intracellular and secreted proteins expression: new methodological developments and applications
Plasmid DNA electrotransfer for intracellular and secreted proteins expression: new methodological developments and applications
复制标题
DOI：
10.1002/jgm.508
复制
发表时间：
2004-02-01
影响因子：
3.5
通讯作者：
Scherman, D
中科院分区：
文献类型：
--
作者：
Bloquel, C;Fabre, E;Scherman, D
体内电转移是一种在各种组织和器官中传递基因的物理方法，它依靠注射质粒DNA然后电脉冲传递。强调了细胞渗透和DNA电泳之间的关系对电传递效率的重要性。体内电转移是最有效的非病毒基因传递策略，并且具有成本低、易于实现和安全等特点，因此备受关注。通过优化质粒在靶器官中的生物分布、质粒结构和编码蛋白的设计，可以进一步提高该技术的潜力。特别是，我们发现较小尺寸的质粒比较大的质粒电转移效率更高。研究和了解转基因基因的动态表达也很重要，这取决于许多因素，包括蛋白质的细胞定位，生理活性和调控。最广泛的目标组织是骨骼肌，因为这种策略不仅有希望治疗肌肉疾病，而且还可以全身分泌治疗性蛋白质。疫苗接种和肿瘤基因治疗也是电转移的主要应用领域，而在肝脏、大脑和角膜等其他器官的应用正在扩大。许多已发表的研究表明，质粒电转移可以对各种疾病如癌症、血液疾病、类风湿关节炎或肌肉缺血产生持久的治疗效果。DNA电转移也是研究特定组织中基因功能的有力实验室工具。版权所有：John Wiley Sons， Ltd. 2004
In vivo electrotransfer is a physical method of gene delivery in various tissues and organs, relying on the injection of a plasmid DNA followed by electric pulse delivery. The importance of the association between cell permeabilization and DNA electrophoresis for electrotransfer efficiency has been highlighted. In vivo electrotransfer is of special interest since it is the most efficient non-viral strategy of gene delivery and also because of its low cost, easiness of realization and safety. The potentiality of this technique can be further improved by optimizing plasmid biodistribution in the targeted organ, plasmid structure, and the design of the encoded protein. In particular, we found that plasmids of smaller size were electrotransferred more efficiently than large plasmids. It is also of importance to study and understand kinetic expression of the transgene, which can be very variable, depending on many factors including cellular localization of the protein, physiological activity and regulation. The most widely targeted tissue is skeletal muscle, because this strategy is not only promising for the treatment of muscle disorders, but also for the systemic secretion of therapeutic proteins. Vaccination and oncology gene therapy are also major fields of application of electrotransfer, whereas application to other organs such as liver, brain and cornea are expanding. Many published studies have shown that plasmid electrotransfer can lead to long-lasting therapeutic effects in various pathologies such as cancer, blood disorders, rheumatoid arthritis or muscle ischemia. DNA electrotransfer is also a powerful laboratory tool to study gene function in a given tissue. Copyright (C) 2004 John Wiley Sons, Ltd.