期刊论文

Construction and Function Analysis of Artificial miRNA Targeting to HIV-1 CCR5 Gene

中文

中国皮肤性病学杂志

1001-7089

2017

31

4

359-364

10.13735/j.cjdv.1001-7089.201608034

湖南省自然科学基金重点项目（09JJ3034）；

本校为第一机构

目的构建人工miR-CCR5,并研究其对HIV-1感染宿主细胞的影响。方法以miR-155为基础骨架,根据HIV-1 CCR5基因序列设计并合成4对miRNA寡聚单链DNA,构建4个人工miR-CCR5。采用qPCR技术检测其对CCR5基因的沉默效率和对干扰素表达的影响;MTT法测定其对被转染细胞的毒性;抗HIV-1_(BaL)感染实验检测其对HIV-1感染的抑制作用。结果qPCR结果显示,4个人工miR-CCR5对CCR5基因都具有一定的沉默效果,其中miR-CCR54的沉默效率最高,达72% ;且不会对细胞干扰素的表达产生影响。MTT实验证实miR-CCR5-4不会影响被转染细胞的活性,此外,病毒抑制实验显示,miR-CCR5-4对HIV-1_(BaL) p24蛋白的抑制作用达70.48%,效果明显...

Objective To construct artificial miR-CCR5 and study its effects on HIV-1 infected host cells. Methods Four pairs of miRNA oligonucleotides were designed and synthesized based on HIV-1 CCR5 gene sequences to construct four artificial miR-CCR5,which derived from the native miR-CCR5 gene sequence. Then qPCR was used to investigate the gene silence efficiency and the expression of IFN-β gene; the cell toxicity was tested by MTT assays and HIV-1 infection was analyzed by detecting p24 protein content. Results The qPCR results showed that the four ...