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Cited by 584RNA Interference and Gene DeliveryMicroRNA in disease regulationPlant Molecular Biology Research

Sequence-specific inhibition of small RNA function

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Hutvágner, György, Simard, Martin J, Mello, Craig C, Zamore, Phillip D·2004·PLoS Biology
DOI 10.1371/journal.pbio.0020098PMC PMC350664
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Abstract

Hundreds of microRNAs (miRNAs) and endogenous small interfering RNAs (siRNAs) have been identified from both plants and animals, yet little is known about their biochemical modes of action or biological functions. Here we report that 2'-O-methyl oligonucleotides can act as irreversible, stoichiometric inhibitors of small RNA function. We show that a 2'-O-methyl oligonucleotide complementary to an siRNA can block mRNA cleavage in Drosophila embryo lysates and HeLa cell S100 extracts and in cultured human HeLa cells. In Caenorhabditis elegans, injection of the 2'-O-methyl oligonucleotide complementary to the miRNA let-7 can induce a let-7 loss-of-function phenocopy. Using an immobilized 2'-O-methyl oligonucleotide, we show that the C. elegans Argonaute proteins ALG-1 and ALG-2, which were previously implicated in let-7 function through genetic studies, are constituents of a let-7-containing protein-RNA complex. Thus, we demonstrate that 2'-O-methyl RNA oligonucleotides can provide an efficient and straightforward way to block small RNA function in vivo and furthermore can be used to identify small RNA-associated proteins that mediate RNA silencing pathways.

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  • Methods have been developed to manipulate miRNA function pharmacologically, facilitating the development of new cancer therapeutic strategies.

    “Methods have been developed to manipulate miRNA function pharmacologically, facilitating the development of new cancer therapeutic strategies (Hutvagner et al., 2004; Krutzfeldt et al., 2005).”

    From Modulation of K-Ras-Dependent Lung Tumorigenesis by MicroRNA-21 (2010) · cites this paper as Hutvagner et al., 2004

    The Results and Discussion report that 2′-O-methyl oligonucleotides inhibit specific miRNAs and can be used to study their function in C. elegans. The paper presents them as a research tool; it does not establish that they facilitate new cancer therapeutic strategies. (LLM, conf=0.87)

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Cited by

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Authors
Hutvágner, G., Simard, M. J., Mello, C. C., & Zamore, P. D.
Affiliations
  • University of Massachusetts Chan Medical School (US)
  • Howard Hughes Medical Institute (US)
ORCID
Venue
PLoS Biology
Publisher
Public Library of Science (PLoS)
Published
2004-02-24
Volume
2
Issue
4
Pages
e98
ISSN
  • 1544-9173 (ISSN-L)
  • 1545-7885
DOI
10.1371/journal.pbio.0020098
PMC
PMC350664
OpenAlex
W4241434797
Cited by
584
References
2
Citations by year
2026: 1 · 2025: 1 · 2024: 10 · 2023: 8 · 2022: 13 · 2021: 9 · 2020: 15 · 2019: 12 · 2018: 22 · 2017: 30 · 2016: 28 · 2015: 21 · 2014: 36 · 2013: 32 · 2012: 44
FWCI
17.09
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS Biology (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • eScholarship@UMassChan (submittedVersion)
  • eScholarship@UMassChan (submittedVersion, cc-by)
  • Discovery Research Portal (University of Dundee) (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • UTS ePRESS (University of Technology Sydney) (submittedVersion)
Funders
  • W. M. Keck Foundation
  • Charles A. King Trust
  • National Institutes of Health (GM62862-01, GM62862, GM58800)
  • Canadian Institutes of Health Research
  • National Institute of General Medical Sciences (GM-58800, R37GM058800, R01GM062862, GM-65236-01, R01GM065236, GM62862-01, R01GM058800, R37GM062862)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Molecular Biology Research
Keywords
2'-O-methyl oligonucleotides, microRNA, endogenous small interfering RNAs, RNA silencing, let-7 microRNA, Argonaute proteins, ALG-1, ALG-2, protein-RNA complex, mRNA cleavage, Caenorhabditis elegans, HeLa cells
MeSH terms
Animals, Drosophila, Genetic Techniques, HeLa Cells, Humans, Molecular Sequence Data, Oligonucleotides, Phenotype, RNA, Messenger, Transfection, Blotting, Northern, Caenorhabditis elegans, Biotinylation, Gene Silencing, Animals, Genetically Modified, RNA Interference, RNA, Small Interfering, MicroRNAs

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