Abstract
Short interfering RNAs (siRNAs) are double-stranded RNAs of approximately 21-25 nucleotides that have been shown to function as key intermediaries in triggering sequence-specific RNA degradation during posttranscriptional gene silencing in plants and RNA interference in invertebrates. siRNAs have a characteristic structure, with 5'-phosphate/3'-hydroxyl ends and a 2-base 3' overhang on each strand of the duplex. In this study, we present data that synthetic siRNAs can induce gene-specific inhibition of expression in Caenorhabditis elegans and in cell lines from humans and mice. In each case, the interference by siRNAs was superior to the inhibition of gene expression mediated by single-stranded antisense oligonucleotides. The siRNAs seem to avoid the well documented nonspecific effects triggered by longer double-stranded RNAs in mammalian cells. These observations may open a path toward the use of siRNAs as a reverse genetic and therapeutic tool in mammalian cells.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Caenorhabditis elegans / genetics
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Caenorhabditis elegans / metabolism
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Caenorhabditis elegans Proteins*
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Calmodulin-Binding Proteins / biosynthesis
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Calmodulin-Binding Proteins / genetics
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Cell Death
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Cells, Cultured / drug effects
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Cells, Cultured / metabolism
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Chloramphenicol O-Acetyltransferase / biosynthesis
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Chloramphenicol O-Acetyltransferase / genetics
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Endoribonucleases / metabolism
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Gene Silencing / drug effects
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Gene Silencing / physiology*
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Genes, Reporter
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Green Fluorescent Proteins
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HeLa Cells / drug effects
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HeLa Cells / metabolism
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Helminth Proteins / biosynthesis
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Helminth Proteins / genetics
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Humans
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Invertebrates / genetics*
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Luminescent Proteins / biosynthesis
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Luminescent Proteins / genetics
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Mammals / genetics
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Mice
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Muscle Proteins / biosynthesis
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Muscle Proteins / genetics
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Phosphorylation
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RNA, Antisense / chemical synthesis
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RNA, Antisense / pharmacology
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RNA, Double-Stranded / pharmacology
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RNA, Double-Stranded / physiology*
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RNA, Helminth / physiology*
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Recombinant Fusion Proteins / biosynthesis
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Ribonuclease III
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Species Specificity
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Transfection
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Vertebrates / genetics*
Substances
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Caenorhabditis elegans Proteins
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Calmodulin-Binding Proteins
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Helminth Proteins
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Luminescent Proteins
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Muscle Proteins
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RNA, Antisense
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RNA, Double-Stranded
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RNA, Helminth
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Recombinant Fusion Proteins
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unc-22 protein, C elegans
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Green Fluorescent Proteins
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Chloramphenicol O-Acetyltransferase
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Endoribonucleases
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Ribonuclease III