A large-scale chemical modification screen identifies design rules to generate siRNAs with high activity, high stability and low toxicity

The use of chemically synthesized short interfering RNAs (siRNAs) is currently the method of choice to manipulate gene expression in mammalian cell culture, yet improvements of siRNA design is expectably required for suc
The use of chemically synthesized short interfering RNAs (siRNAs) is currently the method of choice to manipulate gene expression in mammalian cell culture, yet improvements of siRNA design is expectably required for successful application in vivo. Several studies have aimed at improving siRNA performance through the introduction of chemical modifications but a direct comparison of these results is difficult. We have directly compared the effect of 21 types of chemical modifications on siRNA activity and toxicity in a total of 2160 siRNA duplexes. We demonstrate that siRNA activity is primarily enhanced by favouring the incorporation of the intended antisense strand during RNA-induced silencing complex (RISC) loading by modulation of siRNA thermodynamic asymmetry and engineering of siRNA 3-overhangs. Collectively, our results provide unique insights into the tolerance for chemical modifications and provide a simple guide to successful chemical modification of siRNAs with improved activity, stability and low toxicity.
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Author:Jesper B. Bramsen, Maria B. Laursen, Anne F. Nielsen, Thomas B. Hansen, Claus Bus, Niels Langkjaer, B. Ravindra Babu, Torben Hojland, Mikhail Abramov, Arthur Van Aerschot, Dalibor Odadžić, Romualdas Smicius, Jens Haas, Cordula Andree, Jharna Barman, Malgorzata Wenska, Puneet Srivastava, Chuanzheng Zhou, Dmytro Honcharenko, Simone Hess, Elke Müller, Georgii V. Bobkov, Sergey N. Mikhailov, Eugenio Fava, Thomas F. Meyer, Jyoti Chattopadhyaya, Marino Zerial, Joachim W. Engels, Piet Herdewijn, Jesper Wengel, Jørgen Kjems
URN:urn:nbn:de:hebis:30-74453
Document Type:Article
Language:English
Date of Publication (online):2010/01/27
Year of first Publication:2009
Publishing Institution:Univ.-Bibliothek Frankfurt am Main
Release Date:2010/01/27
Tag:IN-VIVO ; LOCKED NUCLEIC-ACID ; MAMMALIAN-CELLS ; MODIFIED OLIGONUCLEOTIDES ; PASSENGER-STRAND ; SMALL INTERFERING RNA ; STRUCTURAL BASIS
Note:
© 2009 The Author(s) This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Source:Nucleic Acids Research, 37, No. 9, S. 2867-2881 ; doi:10.1093/nar/gkp106
HeBIS PPN:221282807
Institutes:Biochemie und Chemie
Dewey Decimal Classification:570 Biowissenschaften; Biologie
Sammlungen:Universitätspublikationen
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Licence (German):License Logo Veröffentlichungsvertrag für Publikationen

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