In vivo non-invasive monitoring of dystrophin correction in a new Duchenne muscular dystrophy reporter mouse
In vivo non-invasive monitoring of dystrophin correction in a new Duchenne muscular dystrophy reporter mouse
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DOI：
10.1038/s41467-019-12335-x
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发表时间：
2019-10-04
影响因子：
16.6
通讯作者：
Olson, Eric N.
中科院分区：
文献类型：
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作者：
Amoasii, Leonela;Li, Hui;Olson, Eric N.
杜氏肌营养不良症（DMD）是一种由肌营养不良蛋白基因突变引起的致命性遗传病。为了能够在体内无创分析DMD基因校正策略，我们在小鼠体内引入了含有抗肌营养不良蛋白基因c端的荧光素酶报告基因。该报告基因的表达模仿内源性肌营养不良蛋白的表达，DMD突变破坏肌营养不良蛋白的开放阅读框，从而抑制荧光素酶的表达。我们评估了缺失外显子50（一个常见的突变热点）的小鼠在用腺相关病毒传递CRISPR/Cas9基因编辑机制后，对与荧光素酶偶联的肌营养不良蛋白阅读框的纠正。生物发光监测显示，在受影响的骨骼肌和心脏中，肌营养不良蛋白表达的有效和快速恢复。我们的研究结果为监测DMD小鼠模型中的肌营养不良蛋白纠正提供了一种敏感的非侵入性手段，并为测试改善DMD发病机制的不同策略提供了一个平台。
Duchenne muscular dystrophy (DMD) is a fatal genetic disorder caused by mutations in the dystrophin gene. To enable the non-invasive analysis of DMD gene correction strategies in vivo, we introduced a luciferase reporter in-frame with the C-terminus of the dystrophin gene in mice. Expression of this reporter mimics endogenous dystrophin expression and DMD mutations that disrupt the dystrophin open reading frame extinguish luciferase expression. We evaluated the correction of the dystrophin reading frame coupled to luciferase in mice lacking exon 50, a common mutational hotspot, after delivery of CRISPR/Cas9 gene editing machinery with adeno-associated virus. Bioluminescence monitoring revealed efficient and rapid restoration of dystrophin protein expression in affected skeletal muscles and the heart. Our results provide a sensitive non-invasive means of monitoring dystrophin correction in mouse models of DMD and offer a platform for testing different strategies for amelioration of DMD pathogenesis.