In Vivo Modeling of Skeletal Muscle Diseases Using the CRISPR/Cas9 System in Rats
In Vivo Modeling of Skeletal Muscle Diseases Using the CRISPR/Cas9 System in Rats
复制标题
使用 CRISPR/Cas9 系统在大鼠体内建立骨骼肌疾病模型
DOI：
10.1007/978-1-0716-3036-5_20
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Yamanouchi Keitaro
中科院分区：
文献类型：
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作者：
Nakamura Katsuyuki;Tanaka Takao;Yamanouchi Keitaro
CRISPR/Cas9系统是一种强大的基因编辑工具，可用于修饰几乎所有物种的靶基因。它开启了在除小鼠以外的实验室动物中产生敲除或敲入基因的可能性。Dystrophin基因与人杜氏肌营养不良症有关;然而，与人类相比，Dystrophin基因突变小鼠不显示严重的肌肉退化表型。另一方面，用CRISPR/Cas9系统制造的Dystrophin基因突变大鼠表现出比小鼠更严重的表型。在肌营养不良蛋白突变大鼠中观察到的表型更能代表人DMD的特征。这意味着大鼠是比小鼠更好的人类骨骼肌疾病模型。在本章中，我们提出了一个详细的方案，通过使用CRISPR/Cas9系统显微注射到胚胎中来产生基因修饰的大鼠。
The CRISPR/Cas9 system is a powerful gene editing tool that can be used to modify a target gene in almost all species. It unlocks the possibility of generating knockout or knock-in genes in laboratory animals other than mice. The Dystrophin gene is implicated in human Duchenne muscular dystrophy; however, Dystrophin gene mutant mice do not show severe muscle degenerating phenotypes when compared to humans. On the other hand, Dystrophin gene mutant rats made with the CRISPR/Cas9 system show more severe phenotypes than those seen in mice. The phenotypes seen in dystrophin mutant rats are more representative of the features of human DMD. This implies that rats are better models of human skeletal muscle diseases than mice. In this chapter, we present a detailed protocol for the generation of gene-modified rats by microinjection into embryos using the CRISPR/Cas9 system.