The deletion of mutant SOD1 via CRISPR/Cas9/sgRNA prolongs survival in an amyotrophic lateral sclerosis mouse model.,Gene Therapy

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The deletion of mutant SOD1 via CRISPR/Cas9/sgRNA prolongs survival in an amyotrophic lateral sclerosis mouse model.
Gene Therapy ( IF 4.6 ) Pub Date : 2019-12-09 , DOI: 10.1038/s41434-019-0116-1
Weisong Duan _{1,

2,

3} , Moran Guo _{1,

2} , Le Yi _{1,

2} , Yakun Liu _{1,

2} , Zhongyao Li _{1,

2,

3} , Yanqin Ma ₄ , Guisen Zhang ₄ , Yaling Liu _{1,

2,

3} , Hui Bu _{1,

2,

3} , Xueqin Song _{1,

2,

3} , Chunyan Li _{1,

2,

3}

Affiliation

The superoxide dismutase 1 (SOD1) mutation is one of the most notable causes of amyotrophic lateral sclerosis (ALS), and modifying the mutant SOD1 gene is the best approach for the treatment of patients with ALS linked to the mutations in this gene. Clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas9)/sgRNA delivered by the adeno-associated virus (AAV) system is a powerful tool for genome editing in the central nervous system (CNS). Here, we tested the capacity of the AAV-SaCas9-sgRNA system to modify mutant SOD1 in SOD1G93A transgenic mice and found that AAV9-SaCas9-sgRNA5 deleted the SOD1 gene, improved the lifespan of SOD1G93A mice by 54.6%, and notably ameliorated the performance of ALS transgenic mice. An immunochemical analysis showed that the expression of mutant SOD1 was very weak in motor neurons expressing SaCas9-sgRNA5. Consequently, the area showing muscle atrophy was more notably restored in the group treated with SaCas9-sgRNA5 compared with the group treated with SaCas9-sgLacZ. In addition, deep sequencing did not show the indel mutation in the gene highly matched to sgRNA5. Hence, AAV9-SaCas9-sgRNA-based gene editing is a feasible potential treatment for patients with ALS linked to SOD1 mutations.

中文翻译：

通过 CRISPR/Cas9/sgRNA 删除突变型 SOD1 可延长肌萎缩侧索硬化症小鼠模型的存活时间。

超氧化物歧化酶 1 (SOD1) 突变是肌萎缩侧索硬化症 (ALS) 的最显着原因之一，修饰突变的 SOD1 基因是治疗与该基因突变相关的 ALS 患者的最佳方法。由腺相关病毒 (AAV) 系统传递的成簇规则间隔短回文重复 (CRISPR)/CRISPR 相关 (Cas9)/sgRNA 是中枢神经系统 (CNS) 基因组编辑的强大工具。在这里，我们测试了AAV-SaCas9-sgRNA系统在SOD1G93A转基因小鼠中修饰突变SOD1的能力，发现AAV9-SaCas9-sgRNA5删除了SOD1基因，使SOD1G93A小鼠的寿命提高了54.6%，并且显着改善了性能ALS 转基因小鼠。免疫化学分析表明，表达SaCas9-sgRNA5的运动神经元中突变体SOD1的表达非常弱。因此，与用SaCas9-sgLacZ治疗的组相比，用SaCas9-sgRNA5治疗的组中显示肌肉萎缩的区域更显着地恢复。此外，深度测序并未显示与sgRNA5高度匹配的基因存在indel突变。因此，基于 AAV9-SaCas9-sgRNA 的基因编辑对于与 SOD1 突变相关的 ALS 患者来说是一种可行的潜在治疗方法。

更新日期：2019-12-09

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