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  • 李昊,师建辉,魏纯纯,麻献华,陈玉霞,章卫平*.碳水化合物反应元件结合蛋白条件性基因敲除小鼠模型的建立及鉴定[J].第二军医大学学报,2019,40(2):142-148    [点击复制]
  • LI Hao,SHI Jian-hui,WEI Chun-chun,MA Xian-hua,CHEN Yu-xia,ZHANG Wei-ping*.Generation and identification of carbohydrate response element-binding protein conditional gene knockout mouse model[J].Acad J Sec Mil Med Univ,2019,40(2):142-148   [点击复制]
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碳水化合物反应元件结合蛋白条件性基因敲除小鼠模型的建立及鉴定
李昊,师建辉,魏纯纯,麻献华,陈玉霞,章卫平*
0
(海军军医大学(第二军医大学)基础医学院病理生理学教研室, 上海 200433
*通信作者)
摘要:
目的 建立转录因子碳水化合物反应元件结合蛋白(ChREBP)的条件性基因敲除小鼠模型,为研究ChREBP的体内生物学功能提供技术手段。方法和结果 利用Cre/loxP基因打靶策略,通过构建基因打靶载体和基于胚胎干细胞(ES细胞)的基因同源重组,在ChREBP基因第8外显子的两侧分别引入loxP位点;将打靶成功的ES细胞显微注射至小鼠囊胚,然后植入假孕雌鼠子宫获得嵌合鼠,进一步获得可种系传代的ChREBPflox/+小鼠;将ChREBPflox/+小鼠与Alb-Cre小鼠交配,获得ChREBP的肝脏特异性基因敲除小鼠。结论 建立了ChREBP条件性基因敲除小鼠模型,为揭示不同组织中ChREBP的生理功能和病理学意义提供了重要手段。
关键词:  碳水化合物反应元件结合蛋白  果糖代谢  脂代谢  基因敲除小鼠  基因打靶
DOI:10.16781/j.0258-879x.2019.02.0142
投稿时间:2018-10-11修订日期:2018-12-25
基金项目:国家自然科学基金重点项目(31730042).
Generation and identification of carbohydrate response element-binding protein conditional gene knockout mouse model
LI Hao,SHI Jian-hui,WEI Chun-chun,MA Xian-hua,CHEN Yu-xia,ZHANG Wei-ping*
(Department of Pathophysiology, College of Basic Medical Sciences, Naval Medical University(Second Military Medical University), Shanghai 200433, China
*Corresponding author)
Abstract:
Objective To establish a conditional knockout mouse model of transcription factor carbohydrate response element-binding protein (ChREBP), so as to provide a technical approach for exploring the biological function of ChREBP in vivo. Methods and results Using Cre/loxP gene targeting strategy, we constructed the gene targeting vector and franked the 8th exon of ChREBP gene with loxP site in embryonic stem cells (ES cells) via homologous recombination. We microinjected the recombined ES clones into blastocysts and implanted into the uterus of pseudopregnant mice to obtain the chimeric mice and ChREBPflox/+ mice. By crossbreeding ChREBPflox/+ mice with Alb-Cre mice, we generated liver-specific ChREBP knockout mice. Conclusion The generation of ChREBP conditional knockout mouse model provides an important means to reveal the physiological function and pathological significance of ChREBP in different tissues.
Key words:  carbohydrate response element-binding protein  fructose metabolism  lipid metabolism  knockout mice  gene targeting