Ago-2-Mediated Slicer Activity Is Essential for Anti-Flaviviral Efficacy of RNAi

被引：10

作者：

Chen, Shuiping ^{[1
]}

Chahar, Harendra S. ^{[1
]}

Abraham, Sojan ^{[1
]}

Wu, Haoquan ^{[1
]}

Pierson, Theodore C. ^{[2
]}

Wang, Xiaozhong A. ^{[3
]}

Manjunath, N. ^{[1
]}

机构：

[1] Texas Tech Univ Hlth Sci Ctr, Dept Biomed Sci, Ctr Excellence Infect Dis Res, Paul L Foster Sch Med, El Paso, TX 79905 USA

[2] NIAID, Viral Pathogenesis Sect, Viral Dis Lab, NIH, Bethesda, MD 20892 USA

[3] Northwestern Univ, Dept Biochem Mol Biol & Cell Biol, Evanston, IL USA

来源：

PLOS ONE | 2011年 / 6卷 / 11期

基金：

美国国家卫生研究院;

关键词：

CELLULAR MICRORNAS; HIV-1; REPLICATION; VIRUS-INFECTION; CODING REGIONS; WEST-NILE; P-BODIES; EXPRESSION; CELLS; INTERFERES; SUPPRESSION;

D O I：

10.1371/journal.pone.0027551

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

RNA interference can be mediated by fully complementary siRNA or partially complementary miRNA. siRNAs are widely used to suppress viral replication and the fully complementary siRNA bound Ago-2 in the RISC is known to degrade the target RNA. Although other argonaute proteins lacking slicer activity can also bind oligonucleotides with both si and miRNA structures, whether they can also contribute to antiviral effects is not entirely clear. We tested si and miRNA structured oligos for target repression in dual luciferase assays as well as for inhibition of Dengue and West Nile virus replication in ES cells expressing individual Ago proteins. In luciferase assays, both fully complementary and partially complementary oligos effectively repressed their targets in all individual Ago expressing cell lines, although the efficacy with fully complementary oligos was higher in Ago-2+ cells. However, partially complementary oligos had no effect on virus replication in any cell line, while fully complementary siRNAs were highly effective in Ago-2 expressing, but not in cells expressing other Ago proteins. This occurred irrespective of whether the target sequences were located in the coding region or 39UTR of the virus. We conclude that Ago-2 slicer activity is essential for anti-viral efficacy of siRNAs and miRNA-mediated translational repression/transcript destabilization is too weak to suppress the abundantly expressed flaviviral proteins.

引用

页数：6

共 38 条

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