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      On the role of RNA amplification in dsRNA-triggered gene silencing.

      Cell

      Transgenes, Animals, Animals, Genetically Modified, Bacterial Proteins, Caenorhabditis elegans, embryology, genetics, Endoribonucleases, physiology, Gene Silencing, Helminth Proteins, Models, Genetic, RNA, Double-Stranded, RNA, Helminth, RNA, Small Interfering, RNA, Untranslated, RNA-Directed DNA Polymerase, Recombinant Fusion Proteins, Ribonuclease III, Sequence Deletion, Transcription Factors

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          Abstract

          We have investigated the role of trigger RNA amplification during RNA interference (RNAi) in Caenorhabditis elegans. Analysis of small interfering RNAs (siRNAs) produced during RNAi in C. elegans revealed a substantial fraction that cannot derive directly from input dsRNA. Instead, a population of siRNAs (termed secondary siRNAs) appeared to derive from the action of a cellular RNA-directed RNA polymerase (RdRP) on mRNAs that are being targeted by the RNAi mechanism. The distribution of secondary siRNAs exhibited a distinct polarity (5'-->3' on the antisense strand), suggesting a cyclic amplification process in which RdRP is primed by existing siRNAs. This amplification mechanism substantially augments the potency of RNAi-based surveillance, while ensuring that the RNAi machinery will focus on expressed mRNAs.

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          11719187

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