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HIV-1 pol基因人工miRNA的构建和功能分析

Construction and function analysis of artificial miRNA targeting to HIV-1 pol gene
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摘要 以miR-155为基础骨架,根据HIV-1 pol基因序列设计并合成4对miRNA寡聚单链DNA,构建4个人工miR-pol,并对其进行功能分析.qPCR结果显示,4个人工miR-pol对pol基因都具有一定的沉默效果,其中miR-pol-4的沉默效率最高,达76%.进一步的实验证实miR-pol-4不会影响被转染细胞的活性,也不会对细胞干扰素的表达产生影响.此外,HIV-1假病毒实验显示,miR-pol-4对HIV-1的复制具有良好的抑制作用.因此,所获得的miR-pol-4将可为进一步的抗HIV-1感染研究提供基础. The stem sequence in the stem- loop structure of the well- characterized native miR- 155 was replaced with siRNA sequences targeting to HIV - 1 pol gene, and four artificial miR - pol were constructed and analyzed. The qPCR results showed that four miR - pol downregulate pol gene expression, and miR - pol - 4 presente the highest gene silence efficiency, reached 76%. The further experiments confirmed that miR -pol -4 has neither effect on the viability of the transfected cells nor interference the expression of IFN - β gene. In addition, it is also verified that miR - pol - 4 could efficiently suppress HIV - 1 replication by using HIV - 1 pseudotyped virus experiment. Therefore, the miR - pol - 4 is a useful candidate for further study on anti - HIV - 1 infection.
作者 王芳宇 何丽芳 孙奉玉 滕涛 张志坚 焦京
机构地区 衡阳师范学院生命科学系 南华大学药学与生命科学学院
出处 《湖南科技大学学报（自然科学版）》 CAS 北大核心 2012年第3期111-116,共6页 Journal of Hunan University of Science And Technology：Natural Science Edition
基金 湖南省教育厅重点项目(07A012) 湖南省自然科学基金重点项目(09JJ3034)
关键词 HIV-1 pol基因 RNA干扰 人工miRNA HIV- 1 pol gene RNAi artificial miRNA
分类号 R373.9 [医药卫生—病原生物学]
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参考文献26

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湖南科技大学学报（自然科学版）
2012年 第3期

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