生物学杂志

生物学杂志 ›› 2025, Vol. 42 ›› Issue (3): 74-.doi: 10.3969/j.issn.2095-1736.2025.03.074

• 研究报告 • 上一篇    下一篇

斑马鱼id4基因在心脏发育功能及其转基因模型构建中的作用

芮宇梦1,2,3， 王宏杰1,2， 何修宴1,2， 吕文超1,2， 祖　尧1,2,3   

  1. 1. 上海海洋大学 国家海洋生物科学国际联合研究中心， 上海 201306；
    2. 上海海洋大学 水产种质资源发掘与利用教育部重点实验室， 上海 201306；
    3. 临港新片区海洋生物医药科技创新型平台， 上海 201306
  • 出版日期:2025-06-18 发布日期:2025-06-17
  • 通讯作者: 祖尧，教授，研究方向为遗传与发育生物学，E-mail:yzu@shou.edu.cn
  • 作者简介:芮宇梦，硕士研究生，研究方向为遗传与发育生物学，E-mail:ym139232@163.com
  • 基金资助:
    国家自然科学基金项目（32170423， 31501166）； 临港海洋生物医药平台项目（ZY-2023-0001）

The role of zebrafish id4 gene in cardiac development and its construction of transgenic model

RUI Yumeng1,2,3, WANG Hongjie1,2, HE Xiuyan1,2, LYU Wenchao1,2, ZU Yao1,2,3   

  1. 1. International Research Center for Marine Bioscience, Ministry of Science and Technology, Shanghai Ocean
    University, Shanghai 201306, China; 2. Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources,
    Ministry of Education, Shanghai Ocean University, Shanghai 201306, China; 3. Marine Biomedical Science and
    Technology Innovation Platform of Lin-gang Special Area, Shanghai 201306, China
  • Online:2025-06-18 Published:2025-06-17

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摘要： 为探究id4基因在斑马鱼早期心脏发育中的作用，挖掘id4基因与心脏发育关键基因之间潜在的调控关系，研究首先预测分析得到id4基因的启动子序列，构建id4启动子载体并进行双萤光素酶报告实验（Luciferase）。结合Tol2转座酶系统构建其转基因斑马鱼鱼系为后续的深入研究提供实验材料。启动子分析结果显示，在启动子克隆区域无甲基化CpG岛，并且有多个转录因子结合位点。双萤光素酶报告基因实验显示，转染id4启动子的293T细胞比空载对照组的萤光素酶活性有7.3倍的激活作用（P<0.0001），表明克隆的id4启动子具有很好的表达活性，并且发现心脏发育关键基因nkx2.5和gata5分别对id4启动子有显著激活和抑制作用。对注射Tol2-id4-EGFP与Tol2转座酶的斑马鱼胚胎进行观察，发现2 dpf和3 dpf时荧光表达在斑马鱼心脏区域。实验结果表明，id4基因在早期心脏发育中存在调控作用，为进一步揭示其分子网络调控机制奠定基础，同时为先天性心脏病的致病机理提供斑马鱼模型。

关键词: id4, 斑马鱼, 心脏发育, 启动子, Tol2转座子

Abstract: To investigate the role of theid4gene in the early heart development in zebrafish, and to explore the potential regulatory relationship between theid4gene and key genes of heart development, this study firstly predicted and analyzed the promoter sequences of theid4gene, constructed theid4promoter vector and carried out the dual luciferase reporter assay (Luciferase). Subsequently, using the Tol2 transposase system, a transgenic zebrafish line was constructed to provide experimental material for further in-depth research. Promoter analysis results showed no methylated CpG islands in the promoter cloning region and multiple transcription factor binding sites were identified. The dual-luciferase reporter gene assay demonstrated that 293T cells transfected with theid4promoter exhibited a 7.3-fold increase in luciferase activity compared with the null control (P<0.0001), suggesting that the clonedid4promoter had a strong expression activity. Additionally, it was found that the key cardiac development genesnkx2.5andgata5significantly activated and inhibited theid4promoter, respectively. Observation of zebrafish embryos injected with Tol2-id4-EGFP and Tol2 transposase revealed fluorescent expression in the zebrafish heart region at 2 and 3 days. Results suggested thatid4gene played a role in early heart development, laying a foundation for further elucidation of its molecular regulatory network and providing a zebrafish model for studying the pathogenesis mechanisms of congenital heart disease.

Key words: id4, zebrafish, heart development, promoter, Tol2 transposon

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