Ago-2-Mediated slicer activity is essential for Anti-Flaviviral efficacy of RNAi

Shuiping Chen, Harendra S. Chahar, Sojan Abraham, Haoquan Wu, Theodore C. Pierson, Xiaozhong A. Wang, N. Manjunath

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

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 3′UTR 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.

Original languageEnglish (US)
Article numbere27551
JournalPLoS One
Volume6
Issue number11
DOIs
StatePublished - Nov 10 2011
Externally publishedYes

Fingerprint

RNA Interference
MicroRNAs
microRNA
virus replication
Viruses
Cells
luciferase
Virus Replication
small interfering RNA
Luciferases
Small Interfering RNA
Assays
Argonaute Proteins
proteins
cell lines
RNA
Cell Line
West Nile virus
Reduced instruction set computing
Proteins

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Chen, S., Chahar, H. S., Abraham, S., Wu, H., Pierson, T. C., Wang, X. A., & Manjunath, N. (2011). Ago-2-Mediated slicer activity is essential for Anti-Flaviviral efficacy of RNAi. PLoS One, 6(11), [e27551]. https://doi.org/10.1371/journal.pone.0027551

Ago-2-Mediated slicer activity is essential for Anti-Flaviviral efficacy of RNAi. / Chen, Shuiping; Chahar, Harendra S.; Abraham, Sojan; Wu, Haoquan; Pierson, Theodore C.; Wang, Xiaozhong A.; Manjunath, N.

In: PLoS One, Vol. 6, No. 11, e27551, 10.11.2011.

Research output: Contribution to journalArticle

Chen, S, Chahar, HS, Abraham, S, Wu, H, Pierson, TC, Wang, XA & Manjunath, N 2011, 'Ago-2-Mediated slicer activity is essential for Anti-Flaviviral efficacy of RNAi', PLoS One, vol. 6, no. 11, e27551. https://doi.org/10.1371/journal.pone.0027551
Chen S, Chahar HS, Abraham S, Wu H, Pierson TC, Wang XA et al. Ago-2-Mediated slicer activity is essential for Anti-Flaviviral efficacy of RNAi. PLoS One. 2011 Nov 10;6(11). e27551. https://doi.org/10.1371/journal.pone.0027551
Chen, Shuiping ; Chahar, Harendra S. ; Abraham, Sojan ; Wu, Haoquan ; Pierson, Theodore C. ; Wang, Xiaozhong A. ; Manjunath, N. / Ago-2-Mediated slicer activity is essential for Anti-Flaviviral efficacy of RNAi. In: PLoS One. 2011 ; Vol. 6, No. 11.
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