Author:
Piao Xuejiao,Meng Dawei,Zhang Xue,Song Qiang,Lv Hailong,Jia Yichang
Abstract
AbstractC9ORF72 GGGGCC repeat expansion is the most common genetic cause for amyotrophic lateral sclerosis and frontotemporal dementia, which generates abnormal DNA and RNA structures and produces toxic proteins. Recently, efficacy of CRISPR/Cas9-mediated editing has been proven in treatment of disease. However, DNA low complexity surrounding C9ORF72 expansion increases the off-target risks. Here we provide a dual-gRNA design outside of the low complexity region which enables us to remove the repeat DNA in a ‘cutting-deletion-fusion’ manner with a high fusion efficiency (50%). Our dual-gRNA design limits off-target effect and does not significantly affect C9ORF72 expression. In neurons carrying patient C9ORF72 expansion, our approach removes the repeat DNA and corrects the RNA foci in vitro and in vivo. Therefore, we conclude that our proof-of-concept design correct C9ORF72 repeat expansion, which may have potential therapeutic value for the patients.
Funder
National Natural Science Foundation of China
Key Project from Chinese Ministry of Science and Technology
National Science Foundation for Young Scholars of China
China Postdoctoral Science Foundation
Publisher
Springer Science and Business Media LLC
Cited by
13 articles.
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