西安交通大学学报（医学版）2017，Vol.38Issue(5)：669-673,5.DOI:10.7652/jdyxb201705009
microRNA-181b调控精神分裂症易感基因EGR3的分子机制
The molecular mechanism of microRNA-181b in regulatingthe schizophrenia susceptibility gene EGR3
摘要
Abstract
Objective To verify whether early growth response-3(EGR3) gene is targeted by microRNA-181b using molecular biology methods so as to provide guidance for the subsequent study on microRNA-181b`s role in the molecular mechanisms of schizophrenia.Methods Bioinformatic methods predicted that EGR3 gene is targeted by microRNA-181b.PCR methods amplified the fragment in EGR3 gene 3`UTR including the putative microRNA-181b binding site.Then the sequence was cloned into the pmirGLO luciferase vector.The DNA sequences of the amplified fragments were identified by restriction enzyme digestion and sequencing, and were consistent with the reference sequence from UCSC.This constructed vector was marked as pmirGLO-EGR3 vector.Finally, the pmirGLO vector, the pmirGLO-EGR3 vector, microRNA-181b mimics and negative control (NC) were divided into 5 groups and transfected into HEK393T cells;the luciferase activity was tested by dual luciferase reporter gene assay.Results The results of restriction enzyme digestion and sequencing demonstrated that the PCR fragmentwas successfully cloned into pmirGLO vector.The transfection results showed that the recombinant plasmid was successful transfected into HEK293T under the fluorescence microscope, with transfection efficiency being about 90％.The results of dual luciferase activity assay demonstrated that microRNA-181b significantly decreased the reporter gene`s activity compared with the NC.Conclusion At the cellular level, the schizophrenia susceptibility gene EGR3 was verified to be targeted by micorRNA-181b, which provides a new clue for the subsequent study on microRNA-181b`s role in the molecular mechanisms of schizophrenia.关键词
精神分裂症/microRNA-181b/EGR3/3`UTR/调控Key words
schizophrenia/microRNA-181b/EGR3/3`UTR/regulation分类
医药卫生引用本文复制引用
张蕊,张天布,范怡伦,张钰洋,杨雪文,马捷..microRNA-181b调控精神分裂症易感基因EGR3的分子机制[J].西安交通大学学报（医学版）,2017,38(5):669-673,5.基金项目
国家自然科学基金项目 (No.31371298 （）
81301151) （）
国家级大学生创新性实验项目(No.GJ201610698087) （No.GJ201610698087）
教育部新世纪优秀人才支持计划(No.NCET-13-0452) Supported by the National Natural Science Foundation of China(No.31371298 and 81301151),the National Innovation Experiment Program for University Students (No.GJ201610698087),and the Program for New Century Excellent Talentsin University (No. NCET-13-0452) （No.NCET-13-0452）
