CRISPR/Cas9-mediated mutation of asparagine-linked glycosylation 13 transcript variant <em>1</em> causes epilepsy in mice

• 作者: Chen Xuejin Fang Qian Li Shangang Li Shengtian Liu Yixun Wu Ji Yu Baoli Zhang Yan Zhu Geng
• 来源期刊: 生物组学研究杂志（英文）
• 关键词: Alg13 CRISPR/Cas9 epilepsy mouse mutant
• 年，卷（期）: 2020,（1）
• 所属期刊栏目: Research Article
• 页码范围: 6-12
• 页数: 7页
• 语种: 中文
• DOI: 10.1097/JBR.0000000000000059

摘要

Objective::The asparagine-linked glycosylation 13 homolog ( Alg13) has been identified as causative for congenital disorders of glycosylation type I with epilepsy. The aim of this study was to determine whether mice carrying a mutated version of Alg13 could be used as a model for epileptic encephalopathies or congenital disorders of glycosylation type I. Methods::A model of epileptic encephalopathy was established in C57BL/6 mice by introducing mutations in Alg13 via the clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) system. All surgical procedures were approved by the Institutional Animal Care and Use Committee of Shanghai Jiao Tong University (A2016084) on October 8, 2016. Results::Mice with 3 different mutations, Alg13 -54nt, Alg13 -5nt and Alg13 -4nt, all of which are located in Alg13 transcript variant 1, were created. The Alg13 -5nt mice exhibited spontaneous seizures similar to patients with Alg13 mutations, suggesting that they could be used as a model for epilepsy. Western blot analysis demonstrated that Alg13 -5nt mice had lower levels of Alg13 expression than wild-type mice. Video observations showed that two of the 17 Alg13 -5nt mice had stage 5 seizures involving jumping and falling, while 12 had stage 3 seizures with head nodding. Conclusion::The Alg13 mouse model provides an outstanding tool for studying epileptic encephalopathies and investigating different aspects of defects in glycosylation or other post-translational modification that cannot be assessed in patients or cell culture systems.