Unc13基因胰岛β细胞条件性敲除小鼠模型的构建和鉴定

doi:10.3969/j.issn.1006-7795.2020.01.003

首都医科大学学报 ›› 2020, Vol. 41 ›› Issue (1): 14-20.doi: 10.3969/j.issn.1006-7795.2020.01.003

• 糖尿病的临床与基础研究 • 上一篇下一篇

Unc13基因胰岛β细胞条件性敲除小鼠模型的构建和鉴定

李奇, 卢晶, 朱晓蓉, 熊枫然, 杨金奎

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市糖尿病研究所, 北京 100730

收稿日期:2019-12-12 出版日期:2020-02-21 发布日期:2020-02-13
通讯作者: 杨金奎 E-mail:jinkui.yang@gmail.com
基金资助:
国家重点研发计划（2017YFC0909600），国家自然科学基金（81800688），首都医科大学附属北京同仁医院科研基金（TRYY-KYJJ-2016-013），北京市属医院科研培育计划（PX2019006）。

Generation and identification of Unc13 gene pancreatic beta cell knockout mice model

Li Qi, Lu Jing, Zhu Xiaorong, Xiong Fengran, Yang Jinkui

Department of Endocrinology, Beijing Tongren Hospital, Capital Medical University, Beijing Key Laboratory of Diabetes Research and Care, Bejing Diabetes Institute, Beijing 100730, China

Received:2019-12-12 Online:2020-02-21 Published:2020-02-13
Supported by:
This study was supported by National Key P&D Program of China (2017YFC0909600);National Natural Science Foundation of China (81800688); Foundation of Beijing Tongren Hospital,Capital Medical University (TRYY-KYJJ-2016-013);Beijing Municipal Administration of Hospitals Incubating Program (PX2019006).

摘要/Abstract

摘要： 目的构建Cre重组酶系统调控的Unc13基因胰岛β细胞条件性敲除小鼠，为Unc13基因在胰岛素分泌中扮演的角色和作用机制的体内实验研究提供更特异的动物模型。方法运用CRISPR/Cas9技术构建Unc13^flox/flox转基因小鼠，将其与胰岛β细胞特异性表达Cre重组酶（Ins2-cre）工具鼠进行杂交，采用聚合酶链反应（polymerase chain reaction，PCR）和测序等方法对其子代小鼠的基因型进行鉴定，并对该基因敲除（knock out，KO）小鼠及其同窝野生（wild type，WT）小鼠的体质量、糖耐量、胰岛素分泌水平进行测定。结果成功构建胰岛β细胞特异性Unc13基因条件性敲除小鼠（以下简称为Unc13 KO鼠）。进一步研究显示Unc13 KO鼠与WT鼠相比，体质量无明显变化，但糖耐量受损，胰岛素第一时相分泌下降。结论成功构建了Unc13基因胰岛β细胞条件敲除小鼠动物模型，为探讨Unc13基因在糖尿病发生、发展中的作用提供研究平台。

关键词: 基因敲除, Cre-LoxP重组酶系统, 胰岛素分泌, Unc13基因

Abstract: Objective To construct Unc13 gene pancreatic beta cell knockout mice by the Cre recombinase system for the study of the role and mechanism of Unc13 in insulin secretion. Methods Unc13flox/flox transgenic mice were constructed by CRISPR/Cas9 technology and hybridized with pancreatic beta cell specific Cre recombinase (Ins2-cre)tool mice. The progeny genotype was identified with PCR and sequencing technology. The knock out (KO) and their wild type (WT) mice were used to measure body weight, glucose tolerance, and insulin secretion level. Results The pancreatic beta cell specific Unc13 gene conditional knockout mice (hereinafter referred to as Unc13 KO mice) were constructed. Further studies showed that Unc13 KO mice had impaired glucose tolerance and decreased first phase insulin secretion but no significant change in body weight compared with WT mice. Conclusion The Unc13 KO mice model was constructed, which provides a research platform for exploring the role of Unc13 gene in the development of diabetes.

Key words: gene knockout, Cre-LoxP recombinase system, insulin secretion, Unc13 gene

中图分类号:

R587

李奇, 卢晶, 朱晓蓉, 熊枫然, 杨金奎. Unc13基因胰岛β细胞条件性敲除小鼠模型的构建和鉴定[J]. 首都医科大学学报, 2020, 41(1): 14-20.

Li Qi, Lu Jing, Zhu Xiaorong, Xiong Fengran, Yang Jinkui. Generation and identification of Unc13 gene pancreatic beta cell knockout mice model[J]. Journal of Capital Medical University, 2020, 41(1): 14-20.

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Unc13基因胰岛β细胞条件性敲除小鼠模型的构建和鉴定

Generation and identification of Unc13 gene pancreatic beta cell knockout mice model

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