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

  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},
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… 

Expression of small interfering RNAs targeted against HIV-1 rev transcripts in human cells

A mammalian Pol III promoter system is described capable of expressing functional double-stranded siRNAs following transfection into human cells and achieving up to 4 logs of inhibition of expression from the HIV-1 DNA.

Gene silencing in mammalian cells by preformed small RNA duplexes.

Gene silencing using micro-RNA designed hairpins.

It is reported that synthetic hairpin RNAs that mimic siRNAs and miRNA precursor molecules can target a gene for silencing, and the mechanism of silencing appears to be through mRNA degradation and not translational repression.

RNA interference by small hairpin RNAs synthesised under control of the human 7S K RNA promoter

It is demonstrated that orientation of the target sequence within the shRNA construct is important for interference, and effective interference also depends on the length and/or structure of the shRNAs.

Modulation of gene expression by RNAi.

The present chapter deals with siRNA design, synthesis, transfection, and readout of efficiency in a mammalian cell culture system.

Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems

Synthetic siRNAs can induce gene-specific inhibition of expression in Caenorhabditis elegans and in cell lines from humans and mice, and seem to avoid the well documented nonspecific effects triggered by longer double-stranded RNAs in mammalian cells.

Modulation of HIV-1 replication by RNA interference

The utility of RNAi for modulating the HIV replication cycle is demonstrated and evidence that genomic HIV-1 RNA, as it exists within a nucleoprotein reverse-transcription complex, is amenable to siRNA-mediated degradation is provided.

RNA interference with small hairpin RNAs transcribed from a human U6 promoter-driven DNA vector.

This study constructed a human U6 promoter-driven mammalian expression vector to produce hairpin double-stranded RNA and transfected this into a human cell line, and was able to knock down the gene expression of an enhanced green fluorescence protein.



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.