Journal Article

Specific Inhibition of Gene Expression by Small Double-Stranded RNAs in Invertebrate and Vertebrate Systems

Parrish, Susan, Imani, Farhad, Morgan, Richard A., Caplen, Natasha J., Fire, Andrew

2001

Overview

Short interfering RNAs (siRNAs) are double-stranded RNAs of ≈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.

• Publisher: National Academy of Sciences,National Acad Sciences
• Subject: Animals / Biological Sciences / Caenorhabditis elegans - genetics / Caenorhabditis elegans - metabolism / Caenorhabditis elegans Proteins / Calmodulin-Binding Proteins - biosynthesis / Calmodulin-Binding Proteins - genetics / Cell Death / Cell lines / Cells, Cultured - drug effects / Cells, Cultured - metabolism / Chloramphenicol O-Acetyltransferase - biosynthesis / Chloramphenicol O-Acetyltransferase - genetics / Delta cells / Double stranded RNA / Endoribonucleases - metabolism / Gene expression / Gene Silencing - drug effects / Gene Silencing - physiology / Genes / Genes, Reporter / Genetics / Green Fluorescent Proteins / HeLa cells / HeLa Cells - drug effects / HeLa Cells - metabolism / Helminth Proteins - biosynthesis / Helminth Proteins - genetics / Humans / Invertebrates / Invertebrates - genetics / Luminescent Proteins - biosynthesis / Luminescent Proteins - genetics / Mammals - genetics / Mice / Muscle Proteins - biosynthesis / Muscle Proteins - genetics / Nucleotides / Phosphorylation / Recombinant Fusion Proteins - biosynthesis / Ribonuclease III / Ribonucleic acid / RNA / RNA, Antisense - chemical synthesis / RNA, Antisense - pharmacology / RNA, Double-Stranded - pharmacology / RNA, Double-Stranded - physiology / RNA, Helminth - physiology / siRNAs / Small interfering RNA / Somatic cells / Species Specificity / Transfection / Vertebrates / Vertebrates - genetics