首都医科大学学报 ›› 2015, Vol. 36 ›› Issue (6): 844-847.doi: 10.3969/j.issn.1006-7795.2015.06.002

• 糖尿病基础与临床研究 • 上一篇    下一篇

基于CRISPR/Cas9研究RFX6对斑马鱼胰腺发育的影响

程呈1,2, 吴谦3, 卢晶1,2, 仇海燕1,2, 李倩1,2, 杨金奎1,2   

  1. 1. 首都医科大学附属北京同仁医院内分泌科, 北京 100730;
    2. 糖尿病防治研究北京市重点实验室, 北京 100730;
    3. 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)。

Effect of RFX6 on pancreas development in zebrafish using CRISPR/Cas9

Cheng Cheng1,2, Wu Qian3, Lu Jing1,2, Qiu Haiyan1,2, Li Qian1,2, Yang Jinkui1,2   

  1. 1. Department of Endocrinology, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China;
    2. Key Laboratory of Diabetes Prevention and Control, Beijing 100730, China;
    3. 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: 目的 构建RFX6基因敲除斑马鱼动物模型及初步探究RFX6对斑马鱼胰腺发育的影响。方法 应用CRISPR/Cas9技术,设计针对靶点的gRNA,诱导靶点突变,经测序鉴定得到突变体。结果 成功构建了RFX6基因敲除斑马鱼动物模型,初步研究发现RFX6纯合突变体生长发育迟滞、体型瘦小、呈现糖尿病消瘦体型并且不能产生后代。结论 RFX6在斑马鱼胰腺的发育中占据重要地位,是控制斑马鱼胰腺发育的关键因子。

关键词: 基因敲除, CRISPR/Cas9, 斑马鱼, 胰腺发育, RFX6

Abstract: Objective To construct the RFX6 knockout zebrafish animal model and preliminarily study the effects of RFX6 on the pancreatic development in zebrafish. Methods CRISPR/Cas9 technology was used to design gRNA against the target and induce mutation, thus the mutants can be obtained by sequencing. Results The RFX6 knockout zebrafish animal model was successfully constructed. Preliminary studies showed that RFX6 homozygous mutants suffered from growth retardation, lean and thin body with diabetes, and could not produce offsprings. Conclusion RFX6 plays an important role and is the key factor in the pancreatic development of zebrafish.

Key words: gene knockout, CRISPR/Cas9, zebrafish, pancreas development, RFX6

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