中国现代医学杂志2017，Vol.27Issue(25)：1-7,7.DOI:10.3969/j.issn.1005-8982.2017.25.001
MicroRNA-29b-3p重组腺相关病毒制备及在大鼠前额叶皮质中的表达研究
Construction of recombinant adeno-associated virus expressing miRNA-29b-3p and its expression in prefrontal cortex of rats
摘要
Abstract
Objective To obtain the recombinant adeno-associated virus expressing miRNA-29b-3p (rAAV-miRNA-29b-3p),and examine the infection effect and expression level of rAAV-miRNA-29b-3p in the prefrontal cortex of rats.Methods The pre-miRNA-29b-3p gene fragment was inserted into AAV expression plasmid to construct a recombinant AAV plasmid.The recombinant AAV plasmid,packaging plasmid and helper plasmid were co-transfected into HEK293 cells by LipofiterTM for rAAV-miRNA-29b-3p packaging.The rAAV-miRNA-29b-3p was harvested from transfected HEK293 cells and purified by a resin column,and the titer of the rAAV-miRNA-29b-3p was tested by qRT-PCR.The rAAV-miRNA-29b-3p was injected to the prefrontal cortex of rats using stereotactic apparatus.The infection effect was observed under immunofluorescence microscope,the expression level of miRNA-29b-3p was detected by qRT-PCR.Results High titer of rAAV-miRNA-29b-3p was obtained after successful packaging.The rAAV-miRNA-29b-3p was purified,the virus titer reached 1.0×1012 vg/ml.Strong green fluorescence and high level of miRNA-29b-3p were detected in the infected prefrontal cortex after 21 days of infection.Conclusions The constructed rAAV-miRNA-29b-3p can greatly increase the expression level of miRNA-29b-3p in the prefrontal cortex of rats,which provides a useful tool for further study of the biological function of miRNA-29b-3p.关键词
腺相关病毒/miRNA-29b-3p/前额叶皮质/大鼠Key words
adeno-associated virus/miRNA-29b-3p/prefrontal cortex/rat分类
医药卫生引用本文复制引用
李茂,冯建国,王晓斌,陈前修,万运强..MicroRNA-29b-3p重组腺相关病毒制备及在大鼠前额叶皮质中的表达研究[J].中国现代医学杂志,2017,27(25):1-7,7.基金项目
国家自然科学基金(No:81271478) （No:81271478）
