生物技术通报 ›› 2021, Vol. 37 ›› Issue (2): 80-87.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0848

• 研究报告 • 上一篇    下一篇

miR-378促进脂质生成相关靶基因鉴定

张廷焕(), 龙熙, 郭宗义, 柴捷()   

  1. 1.重庆市畜牧科学院,重庆 402460
    2.农业部养猪科学重点实验室,重庆 402460
  • 收稿日期:2020-07-09 出版日期:2021-02-26 发布日期:2021-02-26
  • 作者简介:张廷焕,男,硕士研究生,研究方向:猪功能基因组;E-mail: ztinghuan@163.com
  • 基金资助:
    重庆市科研院所绩效激励引导专项(19536);国家自然科学基金面上项目(31771376);重庆市畜牧科学院基本科研业务费专项项目(15428)

miR-378 Promoting Lipogenesis and Identification of Target Genes

ZHANG Ting-huan(), LONG Xi, GUO Zong-yi, CHAI Jie()   

  1. 1. Chongqing Academy of Animal Science,Chongqing 402460
    2. Key Laboratory of Pig Industry Sciences of Ministry of Agriculture,Chongqing 402460
  • Received:2020-07-09 Published:2021-02-26 Online:2021-02-26

摘要:

旨为验证miR-378在脂肪细胞中的功能,及其脂质相关靶基因的筛选和鉴定。利用miR-378类似物转染3T3-L1细胞,验证miR-378在脂肪细胞中的功能;根据靶标位点的保守性以及促脂功能确定miR-378潜在靶基因;采用microRNA pulldown技术验证miR-378与靶基因的靶标关系;运用双荧光素报告基因实验确定miR-378与靶基因的结合位点。结果发现,miR-378可以通过增加脂质合成和减少脂质分解两条途经来促进脂肪细胞中脂质生成,确定了miR-378与Runx1t1、Galnt3、和RAB10的靶标关系以及结合的靶位点。

关键词: miR-378, 脂质生成, microRNA pulldown, 靶基因, 靶位点

Abstract:

The purpose of this study was to investigate the function of miR-378 in adipocytes and to filter and verify its target genes. 3T3-L1 cells were transfected with miR-378 mimic to validate the function of mir-378 in adipocytes. The potential target genes of miR-378 were identified according to the conservation of target sites and lipid promoting function. The relationship between miR-378 and its targets was verified by microRNA pulldown technology,and then the binding sites of miR-378 and its targets were determined by Dual-Luciferase Reporter Assay System. Some important results were found that miR-378 promoted lipogenesis in adipocytes by increasing lipid synthesis and decreasing lipid decomposition,and the relationship between miR-378 and Runx1t1,Galnt3,RAB10 and the binding sites were determined.

Key words: miR-378, lipogenesis, microRNA pulldown, target genes, target sites