Analysis of siRNA-suppressor of gene silencing interactions

Research output: Chapter in Book/Report/Conference proceedingChapter

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
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages331-337
Number of pages7
Volume451
ISBN (Print)9781588298270
DOIs
Publication statusPublished - 2008

Publication series

NameMethods in Molecular Biology
Volume451
ISSN (Print)10643745

Fingerprint

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

Keywords

  • In vitro Drosophila RNA silencing system
  • P19
  • Silencing suppressor
  • SiRNA
  • SiRNA binding

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Lakatos, L., & Burgyán, J. (2008). Analysis of siRNA-suppressor of gene silencing interactions. In Methods in Molecular Biology (Vol. 451, pp. 331-337). (Methods in Molecular Biology; Vol. 451). Humana Press. https://doi.org/10.1007/978-1-59745-102-4_23

Analysis of siRNA-suppressor of gene silencing interactions. / Lakatos, L.; Burgyán, J.

Methods in Molecular Biology. Vol. 451 Humana Press, 2008. p. 331-337 (Methods in Molecular Biology; Vol. 451).

Research output: Chapter in Book/Report/Conference proceedingChapter

Lakatos, L & Burgyán, J 2008, Analysis of siRNA-suppressor of gene silencing interactions. in Methods in Molecular Biology. vol. 451, Methods in Molecular Biology, vol. 451, Humana Press, pp. 331-337. https://doi.org/10.1007/978-1-59745-102-4_23
Lakatos L, Burgyán J. Analysis of siRNA-suppressor of gene silencing interactions. In Methods in Molecular Biology. Vol. 451. Humana Press. 2008. p. 331-337. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-59745-102-4_23
Lakatos, L. ; Burgyán, J. / Analysis of siRNA-suppressor of gene silencing interactions. Methods in Molecular Biology. Vol. 451 Humana Press, 2008. pp. 331-337 (Methods in Molecular Biology).
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