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Science 305 (5689): 1437-1441

Copyright © 2004 by the American Association for the Advancement of Science

Argonaute2 Is the Catalytic Engine of Mammalian RNAi

Jidong Liu,1* Michelle A. Carmell,1,2* Fabiola V. Rivas,1 Carolyn G. Marsden,1 J. Michael Thomson,3 Ji-Joon Song,1 Scott M. Hammond,3 Leemor Joshua-Tor,1 Gregory J. Hannon1{dagger}

Abstract: Gene silencing through RNA interference (RNAi) is carried out by RISC, the RNA-induced silencing complex. RISC contains two signature components, small interfering RNAs (siRNAs) and Argonaute family proteins. Here, we show that the multiple Argonaute proteins present in mammals are both biologically and biochemically distinct, with a single mammalian family member, Argonaute2, being responsible for messenger RNA cleavage activity. This protein is essential for mouse development, and cells lacking Argonaute2 are unable to mount an experimental response to siRNAs. Mutations within a cryptic ribonuclease H domain within Argonaute2, as identified by comparison with the structure of an archeal Argonaute protein, inactivate RISC. Thus, our evidence supports a model in which Argonaute contributes "Slicer" activity to RISC, providing the catalytic engine for RNAi.

1 Cold Spring Harbor Laboratory, Watson School of Biological Sciences, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.
2 Program in Genetics, Stony Brook University, Stony Brook, NY 11794, USA.
3 Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, NC 27599, USA.

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* These authors contributed equally to this work.

{dagger} To whom correspondence should be addressed. E-mail: hannon{at}cshl.edu

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