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。
Objective To construct Unc 13 gene pancreatic beta cell knockout mice by the Cre recombinase system for the study of the role and mechanism of Unc 13 in insulin secretion.Methods Unc 13flox/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 Unc 13 gene conditional knockout mice(hereinafter referred to as Unc 13 KO mice)were constructed.Further studies showed that Unc 13 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 Unc 13 KO mice model was constructed,which provides a research platform for exploring the role of Unc 13 gene in the development of diabetes.
作者
李奇
卢晶
朱晓蓉
熊枫然
杨金奎
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)
出处
《首都医科大学学报》
CAS
北大核心
2020年第1期14-20,共7页
Journal of Capital Medical University
基金
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)~~
