Targeted mRNA degradation by double-stranded RNA in vitro (original) (raw)

1. Thomas Tuschl,
2. Phillip D. Zamore,
3. Ruth Lehmann,
4. David P. Bartel, and
5. Phillip A. Sharp
6. The Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA; Center for Cancer Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 USA; Howard Hughes Medical Institute and The Skirball Institute, New York University Medical Center, New York, New York 10016, USA

Abstract

Double-stranded RNA (dsRNA) directs gene-specific, post-transcriptional silencing in many organisms, including vertebrates, and has provided a new tool for studying gene function. The biochemical mechanisms underlying this dsRNA interference (RNAi) are unknown. Here we report the development of a cell-free system from syncytial blastoderm Drosophila embryos that recapitulates many of the features of RNAi. The interference observed in this reaction is sequence specific, is promoted by dsRNA but not single-stranded RNA, functions by specific mRNA degradation, and requires a minimum length of dsRNA. Furthermore, preincubation of dsRNA potentiates its activity. These results demonstrate that RNAi can be mediated by sequence-specific processes in soluble reactions.

• RNAi
• post-transcriptional gene silencing
• dsRNA

Footnotes

• Present addresses: 2Department of Cellular Biochemistry, Max-Planck-Institute for Biophysical Chemistry, D-37077 Göttingen, Germany; 5Department of Biochemistry and Molecular Biology, University of Massachusetts Medical Center, Worcester, Massachusetts 01655, USA.

• ↵These authors contributed equally to the work.

• ↵Corresponding authors.

• E-MAIL Phillip.Zamore{at}umassmed.edu (PDZ) orttuschl{at}mpibpc.gwdg.de (TT); FAX (617) 258-6768.

• • Received September 20, 1999.
  • Accepted October 28, 1999.

• Cold Spring Harbor Laboratory Press

•