欢迎访问《中国临床药理学与治疗学》杂志官方网站,今天是

中国临床药理学与治疗学 ›› 2021, Vol. 26 ›› Issue (4): 376-381.doi: 10.12092/j.issn.1009-2501.2021.04.003

• 基础研究 • 上一篇    下一篇

利用Cre/loxP系统构建乳腺细胞特异性敲除SENP7基因的小鼠模型

孙红1,侯佳林2,蔡加琴1,庄 捷1,魏晓霞1   

  1. 1福建医科大学省立临床医学院,福建省立医院药学部,福州 350001,福建;2福建医科大学附属协和医院乳腺外科,福州 350001,福建

  • 收稿日期:2020-10-19 修回日期:2021-01-06 出版日期:2021-04-26 发布日期:2021-05-11
  • 通讯作者: 魏晓霞,女,副主任药师,研究方向:临床药理学。 Tel: 0591-88216352 E-mail: xxwei0321@163.com
  • 作者简介:孙红,女,博士,副主任药师,研究方向:表观遗传药理学。 E-mail: sunhong7777@fjmu.edu.cn
  • 基金资助:
    福建省自然科学基金面上项目(2019J01177);福建省科技联合基金(2017Y9067);福建省卫健委-中青年骨干人才项目(2019ZQN-35);北京白求恩公益基金项目(BJBQEKYJJ-B19001CS);福建省立医院创双高项目(2019HSJJ06)

Construction of mammary gland cell-specific SENP7 knockout mice by Cre-loxP system

SUN Hong 1, HOU Jialin 2, CAI Jiaqin 1, ZHUANG Jie 1, WEI Xiaoxia 1   

  1. 1 Department of Pharmacy, Shengli Clinical Medical College of Fujian Medical University, Fujian Provincial Hospital, Fuzhou 350001, Fujian, China; 2 Department of Breast Surgery, Fujian Medical University Union Hospital, Fuzhou 350001, Fujian, China
  • Received:2020-10-19 Revised:2021-01-06 Online:2021-04-26 Published:2021-05-11

摘要: 目的:应用Cre-loxP系统构建乳腺细胞SENP7基因条件性敲除的小鼠模型并进行鉴定。方法:将SENP7flox/+杂合子小鼠进行杂交,PCR法鉴定为SENP7flox/flox纯合子小鼠,再与MMTV-Cre杂合子小鼠进行数代杂交,基因型鉴定并筛选获得MMTV-Cre×SENP7flox/flox小鼠,即实验所需的SENP7基因特异性敲除小鼠。采用Real-Time PCR、Western blot和HE染色鉴定SENP7敲除效果,观察乳腺组织形态。结果:PCR基因扩增筛选出基因型为MMTV-Cre×SENP7flox/flox小鼠。与MMTV-Cre×SENP7+/+小鼠相比,MMTV-Cre×SENP7flox/flox小鼠乳腺SENP7基因的mRNA、蛋白表达水平显著降低,乳腺腺体数量明显减少。 结论:本研究利用Cre-loxP技术成功构建了乳腺特异性敲除SENP7基因的纯合子小鼠,为进一步研究SENP7基因在乳腺肿瘤发生发展中的作用提供了优良的工具。

关键词: SENP7, Cre-loxP系统, MMTV-Cre, 乳腺

Abstract: AIM: To construct and identify the mammary gland cell-specific conditional knockout of SENP7 by the Cre-loxP system.  METHODS: The homozygous SENP7flox/flox mice were generated by crossing SENP7flox/+ mice. Genotypic identification was performed by PCR. SENP7flox/flox mice and MMTV-Cre mice had reproduced and genotyped respectively, then the two strains of mice were crossed. The MMTV-Cre×SENP7flox/flox mice were the required mouse model. Real-Time PCR, Western blot and HE staining were applied to identify the knockout effect of SENP7 and to observe the morphology of mammary gland tissue. RESULTS: PCR analysis selected MMTV-Cre×SENP7flox/flox mice. Compared to the MMTV-Cre×SENP7+/+ mice, expression levels of SENP7 mRNA and protein of mammary glands were significantly lower in MMTV-Cre×SENP7flox/flox mice, and the number of mammary glands was significantly reduced, suggesting the SENP7 knockout effect was obvious. CONCLUSION: The mammary cell-specific deletion of the SENP7 gene is successfully constructed by Cre-loxP system, which provides a novel target for studying the pathogenesis of breast tumors at animal level.

Key words: SENP7, Cre-loxP system, MMTV-Cre, mammary gland

中图分类号: