Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells

@article{Elbashir2001DuplexesO2,
  title={Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells},
  author={Sayda M. Elbashir and Jens Harborth and Winfried Lendeckel and Abdullah Yalçin and Klaus Weber and Thomas Tuschl},
  journal={Nature},
  year={2001},
  volume={411},
  pages={494-498}
}
RNA interference (RNAi) is the process of sequence-specific, post-transcriptional gene silencing in animals and plants, initiated by double-stranded RNA (dsRNA) that is homologous in sequence to the silenced gene. The mediators of sequence-specific messenger RNA degradation are 21- and 22-nucleotide small interfering RNAs (siRNAs) generated by ribonuclease III cleavage from longer dsRNAs. Here we show that 21-nucleotide siRNA duplexes specifically suppress expression of endogenous and… 

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References

SHOWING 1-10 OF 68 REFERENCES

RNA interference is mediated by 21- and 22-nucleotide RNAs.

It is demonstrated that 21- and 22-nt RNA fragments are the sequence-specific mediators of RNAi, and evidence that the direction of dsRNA processing determines whether sense or antisense target RNA can be cleaved by the siRNA-protein complex is provided.

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

It is shown that ‘loss-of-function’ phenotypes can be created in cultured Drosophila cells by transfection with specific double-stranded RNAs, which coincides with a marked reduction in the level of cognate cellular messenger RNAs.

A species of small antisense RNA in posttranscriptional gene silencing in plants.

The 25-nucleotide antisense RNA detected in transgene-induced PTGS is likely synthesized from an RNA template and may represent the specificity determinant of PTGS.

Role for a bidentate ribonuclease in the initiation step of RNA interference

Dicer is a member of the RNase III family of nucleases that specifically cleave double-stranded RNAs, and is evolutionarily conserved in worms, flies, plants, fungi and mammals, and has a distinctive structure, which includes a helicase domain and dualRNase III motifs.

RNA Interference and Small Interfering RNAs

  • T. Tuschl
  • Biology
    Chembiochem : a European journal of chemical biology
  • 2001
This minireview will highlight recent advances in understanding the molecular mechanism of RNAi and its biological function and identify components of the RNAi machinery required for posttranscriptional silencing by cosuppression.

Selective reduction of dormant maternal mRNAs in mouse oocytes by RNA interference.

In mouse oocytes, RNAi provides a suitable and robust approach to study the function of dormant maternal mRNAs and faithfully phenocopies the Mos null mutant.

Specific interference with gene function by double-stranded RNA in early mouse development

It is shown that dsRNA is effective as a specific inhibitor of the function of three genes in the mouse, namely maternally expressed c-mos in the oocyte and zygotically expressed E-cadherin or a GFP transgene in the preimplantation embryo.
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