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Hairpin RNA induces secondary small interfering RNA synthesis and silencing in trans in fission yeast
Author(s) -
Simmer Femke,
Buscaino Alessia,
KosBraun Isabelle C,
Kagansky Alexander,
Boukaba Abdelhalim,
Urano Takeshi,
Kerr Alastair R W,
Allshire Robin C
Publication year - 2010
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1038/embor.2009.273
Subject(s) - rna silencing , fission , rna , gene silencing , yeast , biology , small interfering rna , small hairpin rna , rna interference , rna induced transcriptional silencing , microbiology and biotechnology , trans acting sirna , genetics , physics , gene , nuclear physics , neutron
RNA interference (RNAi) is widespread in eukaryotes and regulates gene expression transcriptionally or post‐transcriptionally. In fission yeast, RNAi is tightly coupled to template transcription and chromatin modifications that establish heterochromatin in cis . Exogenous double‐stranded RNA (dsRNA) triggers seem to induce heterochromatin formation in trans only when certain silencing proteins are overexpressed. Here, we show that green fluorescent protein (GFP) hairpin dsRNA allows production of high levels of Argonaute‐associated small interfering RNAs (siRNAs), which can induce heterochromatin formation at a remote locus. This silencing does not require any manipulation apart from hairpin expression. In cells expressing a ura4 + –GFP fusion gene, production of GFP siRNAs causes the appearance of ura4 siRNAs from the target gene. Production of these secondary siRNAs depends on RNA‐dependent RNA polymerase Rdp1 (RDRP Rdp1 ) function and other RNAi pathway components. This demonstrates that transitivity occurs in fission yeast and implies that RDRP Rdp1 can synthesize RNA from targeted RNA templates in vivo , generating siRNAs not homologous to the hairpin.