Nucleic Acids Research Advance Access published online on April 16, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm024
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Specificity, duplex degradation and subcellular localization of antagomirs
1Laboratory of Metabolic Diseases, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA, 3Alnylam Pharmaceuticals Inc., 300 3rd Street, Cambridge, MA 02142 USA and 4Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA
*To whom correspondence should be addressed. Tel: +41 44 633 4560; Fax: +41 44 633 1051; E-mail: stoffel{at}imsb.biol.ethz.ch
Received November 6, 2006. Revised January 3, 2007. Accepted January 3, 2007.
MicroRNAs (miRNAs) are an abundant class of 20–23-nt long regulators of gene expression. The study of miRNA function in mice and potential therapeutic approaches largely depend on modified oligonucleotides. We recently demonstrated silencing miRNA function in mice using chemically modified and cholesterol-conjugated RNAs termed ‘antagomirs’. Here, we further characterize the properties and function of antagomirs in mice. We demonstrate that antagomirs harbor optimized phosphorothioate modifications, require >19-nt length for highest efficiency and can discriminate between single nucleotide mismatches of the targeted miRNA. Degradation of different chemically protected miRNA/antagomir duplexes in mouse livers and localization of antagomirs in a cytosolic compartment that is distinct from processing (P)-bodies indicates a degradation mechanism independent of the RNA interference (RNAi) pathway. Finally, we show that antagomirs, although incapable of silencing miRNAs in the central nervous system (CNS) when injected systemically, efficiently target miRNAs when injected locally into the mouse cortex. Our data further validate the effectiveness of antagomirs in vivo and should facilitate future studies to silence miRNAs for functional analysis and in clinically relevant settings.
2Present Address: Institute for Molecular Systems Biology, Laboratory of Metabolic Diseases, ETH Zürich, Wolfgang Pauli Strasse 16, CH-8093 Zürich, Switzerland.
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