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An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells

Author

Listed:
  • Scott M. Hammond

    (Genetica, Inc.)

  • Emily Bernstein

    (Graduate Program in Genetics, State University of New York at Stony Brook
    Cold Spring Harbor Laboratory)

  • David Beach

    (Genetica, Inc.
    Wolfson Institute for Biological Sciences, University College London)

  • Gregory J. Hannon

    (Cold Spring Harbor Laboratory)

Abstract

In a diverse group of organisms that includes Caenorhabditis elegans , Drosophila, planaria, hydra, trypanosomes, fungi and plants, the introduction of double-stranded RNAs inhibits gene expression in a sequence-specific manner1,2,3,4,5,6,7. These responses, called RNA interference or post-transcriptional gene silencing, may provide anti-viral defence, modulate transposition or regulate gene expression1,6,8,9,10. We have taken a biochemical approach towards elucidating the mechanisms underlying this genetic phenomenon. Here we show that ‘loss-of-function’ phenotypes can be created in cultured Drosophila cells by transfection with specific double-stranded RNAs. This coincides with a marked reduction in the level of cognate cellular messenger RNAs. Extracts of transfected cells contain a nuclease activity that specifically degrades exogenous transcripts homologous to transfected double-stranded RNA. This enzyme contains an essential RNA component. After partial purification, the sequence-specific nuclease co-fractionates with a discrete, ∼25-nucleotide RNA species which may confer specificity to the enzyme through homology to the substrate mRNAs.

Suggested Citation

  • Scott M. Hammond & Emily Bernstein & David Beach & Gregory J. Hannon, 2000. "An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells," Nature, Nature, vol. 404(6775), pages 293-296, March.
    • Handle: RePEc:nat:nature:v:404:y:2000:i:6775:d:10.1038_35005107
    DOI: 10.1038/35005107
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    Cited by:

    1. Le Thi Truc Linh, 2018. "The Microrna 29 family and its regulation," HO CHI MINH CITY OPEN UNIVERSITY JOURNAL OF SCIENCE - ENGINEERING AND TECHNOLOGY, HO CHI MINH CITY OPEN UNIVERSITY JOURNAL OF SCIENCE, HO CHI MINH CITY OPEN UNIVERSITY, vol. 8(1), pages 18-27.
    2. Arnaud Segers & Joachim Carpentier & Frédéric Francis & Rudy Caparros Megido, 2023. "Gene Silencing of laccase 1 Induced by Double-Stranded RNA in Callosobruchus maculatus (Fabricius 1775) (Coleoptera: Chrysomelidae) Suggests RNAi as a Potential New Biotechnological Tool for Bruchid’s," Agriculture, MDPI, vol. 13(2), pages 1-19, February.
    3. Peike Sheng & Lu Li & Tianqi Li & Yuzhi Wang & Nicholas M. Hiers & Jennifer S. Mejia & Jossie S. Sanchez & Lei Zhou & Mingyi Xie, 2023. "Screening of Drosophila microRNA-degradation sequences reveals Argonaute1 mRNA’s role in regulating miR-999," Nature Communications, Nature, vol. 14(1), pages 1-14, December.

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