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

作　　者：王芳宇[1,2] 何丽芳[1] 孙奉玉[1,2] 滕涛[1] 张志坚[1] 焦京[2] 

机构地区：[1]衡阳师范学院生命科学系,功能金属有机材料湖南省重点实验室,湖南衡阳421008 [2]南华大学药学与生命科学学院,湖南衡阳421001

出　　处：《湖南科技大学学报（自然科学版）》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)

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.

关 键 词：HIV-1 pol基因 RNA干扰 人工miRNA 

分 类 号：R373.9[医药卫生—病原生物学]