实验动物科学 ›› 2022, Vol. 39 ›› Issue (2): 11-16.DOI: 10. 3969 / j. issn. 1006-6179. 2022. 02. 002

• 论著 • 上一篇    下一篇

心肌细胞特异性 Snhg5 过表达转基因小鼠的建立和应用

  

  1. (军事科学院军事医学研究院生命组学研究所,北京 102206) 
  • 收稿日期:2021-06-01 出版日期:2022-04-28 发布日期:2022-05-16
  • 通讯作者: 王剑( 1977—) ,女,研究员,研究方向:心脏稳态和疾病发生机制. E-mail:wangjian7773@ 126. com
  • 作者简介:孔静祎( 1996—) ,女,硕士研究生,研究方向:心脏稳态和疾病发生机制. E-mail:1069022963@ qq. com
  • 基金资助:
    国家自然科学基金( 81970134,91740116) ;国家重点研发计划( 2019YFA0801601)

Establishment and Application of Cardiomyocyte-specific Snhg5 Transgenic Mice

  1. ( State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences ( Beijing) , Beijing Institute of Lifeomics, Beijing 102206, China)
  • Received:2021-06-01 Online:2022-04-28 Published:2022-05-16

摘要: 目的 建立心肌细胞特异性过表达 Snhg5 的转基因小鼠,用于研究 Snhg5 在心脏稳态维持中发挥的功能。方法 通过构建心肌细胞特异性过表达 Snhg5 的转基因载体,利用显微注射导入小鼠的受精卵中,经过胚胎移植获得转基因首建者小鼠。利用实时荧光定量 PCR( Real-time PCR,qRT-PCR) 检测子代小鼠 Snhg5 基因整合情况,并检测 Snhg5 在各个组织中的表达情况,进一步通过形态观察、组织学和分子水平的检测分析转基因小鼠的心脏表型。 结果 成功构建心肌细胞特异性过表达 Snhg5 的转基因载体,利用受精卵显微注射技术获得 Snhg5 转基因首建者小鼠 4 只。通过对子代小鼠各组织中 Snhg5 的表达进行检测,发现 Snhg5 只在心脏组织中特异性地过表达。进一步的表型分析发现,与对照小鼠相比,转基因小鼠在异丙肾上腺素刺激下更容易 发生病理性心脏重塑。结论 成功建立了心肌细胞特异性 Snhg5 过表达的转基因小鼠,为研究 Snhg5 在心脏组织中的功能及病理性心脏重塑的发生机制提供了良好的动物模型。

关键词:  , LncRNA, Snhg5, 心肌细胞, 转基因小鼠, 心脏重塑 

Abstract: Objective To establish a transgenic mouse with cardiomyocyte-specific overexpression of Snhg5.Method A construct that directed the expression of Snhg5 to cardiomyocytes was made. The linearized constructs were introduced into the fertilized eggs by microinjection to obtain the transgenic mice. The integration of Snhg5 gene was detected by PCR. The tissue-specific expression of Snhg5 was confirmed by Real-time PCR. Cardiac phenotypic analysis in control and transgenic mice were performed at the gross morphological, histological and molecular levels. Result A transgenic vector that specifically overexpressing Snhg5 in cardiomyocytes was successfully constructed, and 4 Snhg5 transgenic founder mice were successfully obtained by microinjection of fertilized eggs. Detecting the expression level of Snhg5 in different tissues showed that Snhg5 was upregulated specifically in cardiac tissue. Further phenotypic analysis demonstrated that cardiomyocyte-specific Snhg5 transgenic mice were more prone to pathological cardiac remodeling under isoproterenol ( ISO ) stimulation compared to control mice. Conclusion A cardiomyocyte-specific Snhg5 transgenic mouse was generated successfully, providing an ideal mouse model for studying the function of Snhg5 in cardiac tissue and the mechanism underlying the pathological cardiac remodeling.

Key words:

LncRNA, Snhg5, cardiomyocyte, transgenic mice, cardiac remodeling 

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