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Science. 2009 Aug 7;325(5941):750-3. doi: 10.1126/science.1176325.

C3PO, an endoribonuclease that promotes RNAi by facilitating RISC activation.

Author information

1
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.

Abstract

The catalytic engine of RNA interference (RNAi) is the RNA-induced silencing complex (RISC), wherein the endoribonuclease Argonaute and single-stranded small interfering RNA (siRNA) direct target mRNA cleavage. We reconstituted long double-stranded RNA- and duplex siRNA-initiated RISC activities with the use of recombinant Drosophila Dicer-2, R2D2, and Ago2 proteins. We used this core reconstitution system to purify an RNAi regulator that we term C3PO (component 3 promoter of RISC), a complex of Translin and Trax. C3PO is a Mg2+-dependent endoribonuclease that promotes RISC activation by removing siRNA passenger strand cleavage products. These studies establish an in vitro RNAi reconstitution system and identify C3PO as a key activator of the core RNAi machinery.

PMID:
19661431
PMCID:
PMC2855623
DOI:
10.1126/science.1176325
[Indexed for MEDLINE]
Free PMC Article

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