Jørgensen Louise H, Larochelle Nancy, Orlopp Kristian, Dunant Patrick, Dudley Roy W R, Stucka Rolf, Thirion Christian, Walter Maggie C, Laval Steven H, Lochmüller Hanns
Institute of Human Genetics, University of Newcastle, Newcastle upon Tyne NE13BZ, United Kingdom.
Hum Gene Ther. 2009 Jun;20(6):641-50. doi: 10.1089/hum.2008.162.
Duchenne muscular dystrophy (DMD) is an X-linked, lethal genetic disorder affecting the skeletal muscle compartment, and is caused by mutation(s) in the dystrophin gene. Gene delivery of microdystrophin constructs using adeno-associated virus (AAV) and antisense-mediated exon skipping restoring the genetic reading frame are two of the most promising therapeutic strategies for DMD. Both approaches use microdystrophin proteins either directly as a desired construct for gene delivery, using the capacity-limited AAV vectors, or as the therapeutic outcome of gene splicing. Although functionality of the resulting artificial dystrophin proteins can be predicted in silico, experimental evidence usually obtained in transgenic mice is required before human trials. However, the enormous number of potential constructs makes screening assays for dystrophin protein function in vitro and in vivo highly desirable. Here we present data showing that functionality of microdystrophins can be assessed using relatively simple and fast techniques.
杜氏肌营养不良症（DMD）是一种X连锁的致死性遗传疾病，影响骨骼肌，由肌营养不良蛋白基因的突变引起。使用腺相关病毒（AAV）递送微肌营养不良蛋白构建体以及通过反义介导的外显子跳跃恢复基因阅读框是DMD最有前景的两种治疗策略。这两种方法都直接将微肌营养不良蛋白作为使用容量有限的AAV载体进行基因递送的所需构建体，或者作为基因剪接的治疗结果。尽管可以通过计算机模拟预测所得人工肌营养不良蛋白的功能，但在进行人体试验之前，通常需要在转基因小鼠中获得实验证据。然而，潜在构建体数量众多，因此非常需要在体外和体内对肌营养不良蛋白功能进行筛选测定。在此，我们展示的数据表明，可以使用相对简单和快速的技术评估微肌营养不良蛋白的功能。
