Generation of a monkey with MECP2 mutations by TALEN-based gene targeting

• Published in: Neuroscience bulletin 2014-06, Vol.30 (3), p.381-386
• Main Authors: Liu, Zhen, Zhou, Xue, Zhu, Ying, Chen, Zhi-Fang, Yu, Bin, Wang, Yan, Zhang, Chen-Chen, Nie, Yan-Hong, Sang, Xiao, Cai, Yi-Jun, Zhang, Yue-Fang, Zhang, Chen, Zhou, Wen-Hao, Sun, Qiang, Qiu, Zilong
• Format: Article
• Language: English
• Subjects: Anatomy; Anesthesiology; Animals; Biomedical and Life Sciences; Biomedicine; Endonucleases - genetics; Female; Fertilization in Vitro; Gene Targeting - methods; HEK293 Cells; Human Physiology; Humans; Macaca fascicularis; Male; Methyl-CpG-Binding Protein 2 - genetics; Methyl-CpG-Binding Protein 2 - metabolism; Microinjections; Mutation - genetics; Neurology; Neurosciences; Oocytes; Pain Medicine; RNA, Messenger - metabolism; Transcription Factors - genetics; Transcription Factors - metabolism; Transfection; Zygote - physiology; 基因打靶; 基因突变; 基础; 大脑发育; 灵长类动物; 特异性基因; 猴子; 编辑方法
• ISSN: 1673-7067; 1995-8218
• DOI: 10.1007/s12264-014-1434-8

Gene editing in model organisms has provided critical insights into brain development and diseases. Here, we report the generation of a cynomolgus monkey （Macaca fascicularis） carrying MECP2 mutations using transcription activator-like effector nucleases （TALENs）-mediated gene targeting. After injecting TALENs mRNA into monkey zygotes achieved by in vitro fertilization and embryo transplantation into surrogate monkeys, we obtained one male newborn monkey with an MECP2 deletion caused by frame- shifting mutation in various tissues. The monkey carrying the MECP2 mutation failed to survive after birth, due to either the toxicity of TALENs or the critical requirement of MECP2 for neural development. The level of MeCP2 protein was essentially depleted in the monkey＇s brain. This study demonstrates the feasibility of introducing genetic mutations in non-human primates by site-specific gene-editing methods.