In vivo Electroporation of Skeletal Muscle Fibers in Mice.

Steven J Foltz, Criss H Hartzell, Hyojung J Choo
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Abstract

In vitro models are essential for investigating the molecular, biochemical, and cell-biological aspects of skeletal muscle. Still, models that utilize cell lines or embryonic cells do not fully recapitulate mature muscle fibers in vivo. Protein function is best studied in mature differentiated tissue, where biological context is maintained, but this is often difficult when reliable detection reagents, such as antibodies, are not commercially available. Exogenous expression of tagged proteins in vivo solves some of these problems, but this approach can be technically challenging because either a mouse must be engineered for each protein of interest or viral vectors are required for adequate levels of expression. While viral vectors can infect target cells following local administration, they carry the risk of genome integration that may interfere with downstream analyses. Plasmids are another accessible expression system, but they require ancillary means of cell penetration; electroporation is a simple physical method for this purpose that requires minimal training or specialized equipment. Here, we describe a method for in vivo plasmid expression in a foot muscle following electroporation.

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小鼠骨骼肌纤维的体内电穿孔。
体外模型对于研究骨骼肌的分子、生化和细胞生物学方面至关重要。尽管如此,利用细胞系或胚胎细胞的模型并不能完全再现体内成熟的肌肉纤维。蛋白质功能最好在成熟分化的组织中进行研究,在那里保持生物学背景,但当可靠的检测试剂(如抗体)无法商业化时,这通常很困难。标记蛋白在体内的外源表达解决了其中的一些问题,但这种方法在技术上可能具有挑战性,因为必须针对每种感兴趣的蛋白对小鼠进行工程改造,或者需要病毒载体来获得足够的表达水平。虽然病毒载体可以在局部给药后感染靶细胞,但它们具有基因组整合的风险,可能会干扰下游分析。质粒是另一种可获得的表达系统,但它们需要辅助的细胞渗透手段;电穿孔是一种简单的物理方法,只需要最少的训练或专业设备。在这里,我们描述了一种电穿孔后在足部肌肉中体内表达质粒的方法。
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