Analysis of siRNA-suppressor of gene silencing interactions.

Research output: Contribution to journalArticle

Abstract

RNA silencing is an evolutionarily conserved system that functions as an antiviral mechanism in higher plants and animals. To counteract RNA silencing, viruses evolved silencing suppressors that interfere with siRNA guided RNA silencing pathway. We used the heterologous Drosophila in vitro embryo RNA to analyze the molecular mechanism of suppression of silencing suppressors. We found that different silencing suppressors inhibit the RNA silencing via binding to siRNAs. None of the suppressors affected the activity of preassembled RISC complexes. In contrast, suppressors uniformly inhibited the siRNA-initiated RISC assembly pathway by preventing RNA silencing initiator complex formation. Here, we provide the protocol for the detailed analysis of p19 silencing suppressors of tombusviruses in the heterologous Drosophila in vitro system.

Original languageEnglish
Pages (from-to)331-337
Number of pages7
JournalMethods in molecular biology (Clifton, N.J.)
Volume451
Publication statusPublished - 2008

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Suppressor Genes
Gene Silencing
RNA Interference
Small Interfering RNA
Drosophila
Tombusvirus
RNA Viruses
Antiviral Agents
Embryonic Structures
RNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

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abstract = "RNA silencing is an evolutionarily conserved system that functions as an antiviral mechanism in higher plants and animals. To counteract RNA silencing, viruses evolved silencing suppressors that interfere with siRNA guided RNA silencing pathway. We used the heterologous Drosophila in vitro embryo RNA to analyze the molecular mechanism of suppression of silencing suppressors. We found that different silencing suppressors inhibit the RNA silencing via binding to siRNAs. None of the suppressors affected the activity of preassembled RISC complexes. In contrast, suppressors uniformly inhibited the siRNA-initiated RISC assembly pathway by preventing RNA silencing initiator complex formation. Here, we provide the protocol for the detailed analysis of p19 silencing suppressors of tombusviruses in the heterologous Drosophila in vitro system.",
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