High-efficiency gene transfer into skeletal muscle mediated by electric pulses
High-efficiency gene transfer into skeletal muscle mediated by electric pulses
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DOI：
10.1073/pnas.96.8.4262
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发表时间：
1999-04-13
影响因子：
11.1
通讯作者：
Scherman, D
中科院分区：
文献类型：
--
作者：
Mir, LM;Bureau, MF;Scherman, D
将基因递送至骨骼肌是用于治疗肌肉疾病和用于治疗性蛋白质的全身分泌的有前景的策略。然而，目前的DNA递送技术必须在表达水平和个体间变异性方面进行改进。我们报告非常有效的质粒DNA转移肌纤维中使用方波电脉冲的低场强（小于300 V/cm）和长时间（超过1毫秒）。相反的电渗透诱导的摄取小分子进入肌纤维，质粒DNA必须存在于组织中的电脉冲期间，这表明在电转移过程中的电场对DNA的直接影响。这个I. M。电转移方法在小鼠、大鼠、兔和猴的各种肌肉中将报告基因和治疗基因的表达增加了几个数量级。此外，I.M.电转移强烈地降低了可变性。观察表达的稳定性至少9个月。用编码成纤维细胞生长因子1的pCMV-FGF 1质粒，在经受电脉冲的大多数肌纤维中免疫检测到该蛋白质。肌肉中的DNA电转移可能在基因治疗以及生理学、药理学和发育研究中具有广泛的应用。
Gene delivery to skeletal muscle is a promising strategy for the treatment of muscle disorders and for the systemic secretion of therapeutic proteins. However, present DNA delivery technologies have to be improved with regard to both the level of expression and interindividual variability. We report very efficient plasmid DNA transfer in muscle fibers by using square-wave electric pulses of low field strength (less than 300 V/cm) and of long duration (more than 1 ms). Contrary to the electropermeabilization-induced uptake of small molecules into muscle fibers, plasmid DNA has to be present in the tissue during the electric pulses, suggesting a direct effect of the electric field on DNA during electrotransfer. This i.m. electrotransfer method increases reporter and therapeutic gene expression by several orders of magnitude in various muscles in mouse, rat, rabbit, and monkey. Moreover, i.m. electrotransfer strongly decreases variability. Stability of expression was observed for at least 9 months. With a pCMV-FGF1 plasmid coding for fibroblast growth factor 1, this protein was immunodetected in the majority of muscle fibers subjected to the electric pulses. DNA electrotransfer in muscle may have broad applications in gene therapy and in physiological, pharmacological, and developmental studies.