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

doi:10.3969/j.issn.1006-7795.2015.06.001

首都医科大学学报 ›› 2015, Vol. 36 ›› Issue (6): 839-843.doi: 10.3969/j.issn.1006-7795.2015.06.001

• 糖尿病基础与临床研究 • 上一篇下一篇

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

卢晶¹, 吴谦², 谢荣荣¹, 仇海燕¹, 程呈¹, 杨金奎¹

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

收稿日期:2015-10-21 出版日期:2015-12-21 发布日期:2015-12-18
通讯作者: 杨金奎 E-mail:jinkui.yang@gmail.com
基金资助:
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)。

Construction and preliminary application of BACE2 gene knockout in zebrafish using CRISPR/Cas9

Lu Jing¹, Wu Qian², Xie Rongrong¹, Qiu Haiyan¹, Cheng Cheng¹, Yang Jinkui¹

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

Received:2015-10-21 Online:2015-12-21 Published:2015-12-18
Supported by:
This study was supported by National Natural Science Foundation of China(81270918, 81471014), Natural Science Foundation of Beijing(7131005).

摘要/Abstract

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

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.

Key words: gene knockout, CRISPR/Cas9, zebrafish, β-site APP-cleaving enzyme 2

中图分类号:

卢晶, 吴谦, 谢荣荣, 仇海燕, 程呈, 杨金奎. 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]. 首都医科大学学报, 2015, 36(6): 839-843.

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 Medical University, 2015, 36(6): 839-843.

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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

Construction and preliminary application of BACE2 gene knockout in zebrafish using CRISPR/Cas9

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