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β位淀粉样前体蛋白裂解酶2基因敲除斑马鱼动物模型的构建及其初步应用
引用本文:β位淀粉样前体蛋白裂解酶基因敲除斑马鱼动物模型的构建及其初步应用.β位淀粉样前体蛋白裂解酶2基因敲除斑马鱼动物模型的构建及其初步应用[J].首都医学院学报,2015,36(6):839-843.
作者姓名:β位淀粉样前体蛋白裂解酶基因敲除斑马鱼动物模型的构建及其初步应用
作者单位:1. 首都医科大学附属北京同仁医院内分泌科 糖尿病防治研究北京市重点实验室, 北京 100730;2. 北京大学生命科学院, 北京 100871
基金项目:国家自然科学基金项目(81270918,81471014),北京市自然科学基金重点项目(7131005)。
摘    要:目的 构建β位淀粉样前体蛋白裂解酶2(β-site APP-cleaving enzyme 2,BACE2)基因敲除斑马鱼动物模型并及其初步探索其应用。方法 运用CRISPR/Cas9技术,在斑马鱼中设计针对靶点的gRNA,使之诱导靶点突变,经测序鉴定得到突变体。结果 BACE2纯合突变体在早期部分无法存活,RT-PCR结果显示BACE2基因在斑马鱼的早期发育中持续表达。结论 BACE2基因敲除的斑马鱼动物模型将为进一步研究BACE2基因在斑马鱼胰腺发育中的作用和针对糖尿病的药物靶点筛选提供有效工具。

关 键 词:基因敲除  CRISPR/Cas9  斑马鱼  β位淀粉样前体蛋白裂解酶2  
收稿时间:2015-10-21

Construction and preliminary application of BACE2 gene knockout in zebrafish using CRISPR / Cas9
Lu Jing,Wu Qian,Xie Rongrong,Qiu Haiyan,Cheng Cheng,Yang Jinkui.Construction and preliminary application of BACE2 gene knockout in zebrafish using CRISPR / Cas9[J].Journal of Capital University of Medical Sciences,2015,36(6):839-843.
Authors:Lu Jing  Wu Qian  Xie Rongrong  Qiu Haiyan  Cheng Cheng  Yang Jinkui
Institution:1. Key Laboratory of Diabetes Prevention and Control, Department of Endocrinology, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China;2. Life Sciences, Peking University, Beijing 100871, China
Abstract:Objective To construct β-site APP-cleaving enzyme 2 knockout zebrafish animal model and preliminarily explore its application. Methods By using the CRISPR/Cas9 technology, the gRNA against target in zebrafish was designed to induce mutations;afterwards the mutant was obtained by DNA sequencing. Results BACE2 homozygous mutants were unable to survive in the early part. Results of RT-PCR showed continued BACE2 gene expression in zebrafish early development. Conclusion BACE2 knockout zebrafish animal model may provide an effective tool for further study of the mechanism of pancreas development and drug screening.
Keywords:gene knockout  CRISPR/Cas9  zebrafish  β-site APP-cleaving enzyme 2  
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