条件性敲除软骨组织中FGF9基因小鼠模型的建立

doi:10.19438/j.cjoms.2019.05.005

中国口腔颌面外科杂志 ›› 2019, Vol. 17 ›› Issue (5): 412-417.doi: 10.19438/j.cjoms.2019.05.005

条件性敲除软骨组织中FGF9基因小鼠模型的建立

翁梦佳¹, 章筱悦^2,*, 陈振琦^1,*

1.上海交通大学医学院附属第九人民医院·口腔医学院正畸科,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海 200011;
2.同济大学口腔医学院·附属口腔医院正畸科教研室,上海牙组织修复与再生工程技术研究中心,上海 200072

收稿日期:2019-02-13 出版日期:2019-09-20 发布日期:2020-03-11
通讯作者: 陈振琦,E-mail：orthochen@yeah.net;章筱悦,E-mail：ortho_zhangxy@126.com。*共同通信作者
作者简介:翁梦佳（1992-）,女,在读硕士研究生,E-mail：wengmengjia@sjtu.edu.cn

Generation of Fgf9 conditional knockout mice in cartilage

WENG Meng-jia¹, ZHANG Xiao-yue², CHEN Zhen-qi¹

1. Department of Orthodontics, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology & Shanghai Research of Stomatology. Shanghai 200011;
2. Department of Orthodontics, School &Hospital of Stomatology, Tongji University, Shanghai Engineering Research Center of Tooth Restoration and Regeneration. Shanghai 200072, China

Received:2019-02-13 Online:2019-09-20 Published:2020-03-11

摘要/Abstract

摘要： 目的：利用Cre-LoxP系统建立可调控的软骨组织条件性敲除成纤维细胞生长因子9（fibroblast growth factor,FGF9）小鼠模型。方法：构建针对小鼠Fgf9基因2号外显子的重组载体,电穿孔转染胚胎干细胞（ES细胞）。选择药物G418和Ganc,筛选阳性ES细胞并进行PCR鉴定。利用显微注射将ES细胞注入C57BL/6J小鼠囊胚,移入假孕小鼠子宫,获得含Neo的flox杂合小鼠。该小鼠去Neo后,与ColⅡ-Cre转基因小鼠杂交,并以他莫昔芬诱导,其后代软骨细胞内flox纯合、Cre阳性小鼠的flox区域被敲除。结果：Fgf9基因flox杂合子小鼠无明显异常,可稳定繁殖。该flox小鼠与ColⅡ-Cre转基因小鼠交配后,成功建立条件性敲除软骨组织中Fgf9基因小鼠模型。结论：利用Cre-LoxP系统,可成功建立Fgf9基因条件性敲除小鼠模型,为后续研究FGF9在骨软骨发育及骨关节稳态维持中的功能奠定了基础。

关键词: FGF9, 条件性基因敲除, Col II-Cre转基因小鼠

Abstract: PURPOSE: To establish a conditional knockout mouse model of fibroblast growth factor 9 (FGF9) in cartilage with Cre-LoxP system. METHODS: The recombinant vector of exon 2 of mouse Fgf9 was constructed and transfected into ES cells by electroporation. The positive ES cells were screened by G418 and Ganciclovoir and identified by PCR. ES cells were microinjected into the blastocysts of C57BL/6J mice and then transferred into the uterus of pseudopregnant mice to obtain chimeric mice. Flox region in chondrocytes of flox homozygous and Cre-positive fetuse was knocked out by hybridizing the Neo-free mice with Col II-Cre mice and induced with tamoxifen. RESULTS: Fgf9 Flox heterozygous mice reproduced steadily without obvious abnormality. After mating with Col II-Cre transgenic mice, the Flox mice successfully produced a conditional knockout mouse model of Fgf9 in cartilage tissue. CONCLUSIONS: Conditional knockout mice model of FGF9 was successfully established with Cre-LoxP system, which lays a foundation for further study of the function of FGF9 in osteochondral development and bone and joint homeostasis.

Key words: FGF9, Conditional knockout, Col II-Cre transgenic mice

中图分类号:

R783.5

翁梦佳, 章筱悦, 陈振琦. 条件性敲除软骨组织中FGF9基因小鼠模型的建立[J]. 中国口腔颌面外科杂志, 2019, 17(5): 412-417.

WENG Meng-jia, ZHANG Xiao-yue, CHEN Zhen-qi. Generation of Fgf9 conditional knockout mice in cartilage[J]. China J Oral Maxillofac Surg, 2019, 17(5): 412-417.

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条件性敲除软骨组织中FGF9基因小鼠模型的建立

Generation of Fgf9 conditional knockout mice in cartilage

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