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Nucleic Acids Research 2005 33(10):e92; doi:10.1093/nar/gni091
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Published online 7 June 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Methods Online

RNA interference by mixtures of siRNAs prepared using custom oligonucleotide arrays

Andrew V. Oleinikov*, Jun Zhao and Matthew D. Gray

Seattle Biomedical Research Institute Seattle, WA 98109, USA

*To whom correspondence should be addressed at Seattle Biomedical Research Institute, 307 Westlake Avenue N, Suite 500, Seattle, WA 98109-5219, USA. Tel: +1 206 256 7447; Fax: +1 206 256 7229; Email: andrew.oleinikov{at}sbri.org

Received November 28, 2004. Revised March 10, 2005. Accepted May 18, 2005.

RNA interference (RNAi) is a process in which double-strand RNA (dsRNA) directs the specific degradation of a corresponding target mRNA. The mediators of this process are small dsRNAs, of ~21 bp in length, called small interfering RNAs (siRNAs). siRNAs, which can be prepared in vitro in a number of ways and then transfected into cells, can direct the degradation of corresponding mRNAs inside these cells. Hence, siRNAs represent a powerful tool for studying gene functions, as well as having the potential of being highly specific pharmaceutical agents. Some limitations in using this technology exist because the preparation of siRNA in vitro and screening for siRNAs efficient in RNAi can be expensive and time-consuming processes. Here, we demonstrate that custom oligonucleotide arrays can be efficiently used for the preparation of defined mixtures of siRNAs for the silencing of exogenous and endogenous genes. The method is fast, inexpensive, does not require siRNA optimization and has a number of advantages over methods utilizing enzymatic preparation of siRNAs by digestion of longer dsRNAs, as well as methods based on chemical synthesis of individual siRNAs or their DNA templates.

Present addresses: Jun Zhao, Aaken Laboratories, Inc., 227777 County Road 102, Woodland, CA 95695, USA

Matthew D. Gray, MDG Associates, Inc., 3727 SW Elmgrove Street, Seattle, WA 98126-3418, USA


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