P19 of tomato bushy stunt virus suppresses RNA silencing induced by short hairpin RNA in mammal cells
P19 of tomato bushy stunt virus suppresses RNA silencing induced by short hairpin RNA in mammal cells
复制标题
番茄丛矮病毒P19抑制哺乳动物细胞中短发夹RNA诱导的RNA沉默
DOI：
10.1007/s12250-007-0022-3
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发表时间：
2008-01-13
影响因子：
5.5
通讯作者：
Ai-long H
中科院分区：
文献类型：
--
作者：
Wei-xian C;Juan C;Zhen-zhen Z;Ai-long H
为了对抗寄生宿主的免疫系统，一些病毒编码蛋白质来抑制RNA干扰(RNAi)效应。在本报告中，我们建立了两个易于观察的RNAi系统，效果明显。然后利用这两个系统研究了番茄丛状矮化病毒P19在哺乳动物细胞中抑制RNAi的功能。设计针对绿色荧光蛋白(pshRNA-GFP)和萤火虫荧光素酶(pshRNA-Luc)的短发夹状RNA，分别插入真核转录载体pTZU6+1。将shRNA表达载体与含有P19或不含P19目的基因的载体共转染。荧光显微镜、Western blotting和RT-PCR检测GFP的表达水平。用双荧光素酶检测系统分析荧光素酶的表达水平。本研究设计的pshRNA特异性有效地下调了靶基因的表达，两个基因的表达分别下降了约70%。当P19被导入RNAi系统后，与对照组相比，GFP和荧光素酶的表达基本恢复。成功构建了绿色荧光蛋白和荧光素酶的RNAi系统，证明番茄丛矮病毒P19在哺乳动物细胞中具有对抗shRNA诱导的RNAi效应的能力。
To counteract the immune system in parasitic hosts, some viruses encode proteins to suppress the RNA interference (RNAi) effect. In this report, we established two RNAi systems to be easily observed with strong and obvious effect. The function of the P19 of tomato bushy stunt virus, which suppresses RNAi in mammal cells, was then studied using these two systems. Short hairpin RNAs targeting green fluorescence protein (pshRNA-GFP) and firefly luciferase (pshRNA-luc) were designed and inserted into a eukaryotic transcriptional vector pTZU6+1, respectively. The shRNA expressing vectors were co-transfected with plasmids containing the target gene with or without P19. The GFP expression level was assayed by fluorescence microscopy, Western blotting and RT-PCR. The luciferase expression level was analyzed by the dual-luciferase assay system. pshRNA designed in this study down-regulated the target gene specifically and efficiently, with a decrease of expression of both genes of about 70%, respectively. When P19 was introduced into the RNAi systems, the expression of both GFP and the luciferase were mostly recovered compared with the control groups. The RNAi systems of GFP and luciferase were constructed successfully, demonstrating that P19 of tomato bushy stunt virus has the ability to counteract the RNAi effect induced by shRNA in mammal cells.
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影响因子：
56.9
作者：
Li, HW;Li, WX;Ding, SW
通讯作者：
Ding, SW
DOI：
10.1073/pnas.232434999
发表时间：
2002-11-12
影响因子：
11.1
作者：
Mallory, AC;Reinhart, BJ;Bowman, LH
通讯作者：
Bowman, LH
影响因子：
32.4
作者：
Bennasser, Y;Le, SY;Jeang, KT
通讯作者：
Jeang, KT
影响因子：
64.8
作者：
Elbashir, SM;Harborth, J;Tuschl, T
通讯作者：
Tuschl, T
影响因子：
11.8
作者：
Kasschau, KD;Xie, ZX;Carrington, JC
通讯作者：
Carrington, JC