应用基因编辑技术探索建立中枢神经系统动静脉畸形小鼠模型  

Explore the Establishment of Central Nervous System Arteriovenous Malformation Model in Mice by Gene Editing Technology

作　　者：徐宏治[1] 徐铭[2] 陈峻叡 秦智勇[1] 林小凤[2] 陈衔城[1] 

机构地区：[1]复旦大学附属华山医院神经外科,200040 [2]复旦大学附属华山医院麻醉科,200040

出　　处：《中国临床神经科学》2016年第2期194-198,共5页Chinese Journal of Clinical Neurosciences

基　　金：上海市科学技术委员会实验动物研究领域项目(编号:13140903300);上海市科学技术委员会医学重点项目(编号:15411952200)

摘　　要：目的采用CRISPR/Cas9技术编辑小鼠Alk1基因,探讨条件性敲除Alk1建立中枢神经系统动静脉畸形动物模型的可行性。方法利用CRISPR/Cas9技术在小鼠Alk1基因的特定位点插入两个Lox P序列;经传代并与SM22-Cre小鼠交配后培育出Alk1^(2Lox P/2Lox P)/Cre目标小鼠。该目标小鼠8周龄时通过他莫昔芬腹腔注射激活Cre重组酶的表达,从而条件性敲除Alk1基因诱导脑动静脉畸形的生成。2周后取小鼠脑组织标本,进行苏木精-伊红染色和病理分析。结果 7只目标小鼠中,2只脑组织病理切片发现管腔大小不一的异常血管病灶。结论条件性敲除Alk1基因可初步建立中枢神经系统动静脉畸形动物模型。Aim To create the brain arteriovenous malformation（AVM） models in mice with conditional knock-out of Alk1 gene, facilitated by CRISPR/Cas9 gene editing technology. Methods The CRISPR/Cas9 technology was used to insert two LoxP sequences into the target sites of Alk1 in mice, which were intercrossed with SM22-Cre mice to ultimately generate Alk12 LoxP/2LoxP/Cre target mice. The target mice at the age of 8 weeks were given tamoxifen intraperitoneally to induce Cre expression, so as to conditionally knock out Alk1 flanked by LoxP. Two weeks later, the brain specimens were taken and the AVM lesions were pathologically analyzed. Results Among the 7 target mice obtained, there were 2 mice in which the brain tissue slides showed typical AVM-like abnormal vessels in various sizes. Conclusion Conditional knock-out of the Alk1 gene in postnatal mice is a feasible way to create brain AVM mice models.

关 键 词：CRISPR/Cas9基因编辑技术 Alk1 脑动静脉畸形 模型 小鼠 

分 类 号：Q189[生物学—神经生物学] R743[生物学—普通生物学]