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Nucleic Acids Research 2006 34(5):e36; doi:10.1093/nar/gnj028
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Published online 3 March 2006

© The Author 2006. 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@oxfordjournals.org

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Co-expression of anti-NF{kappa}B RNA aptamers and siRNAs leads to maximal suppression of NF{kappa}B activity in mammalian cells

Robert Chan, Madaline Gilbert, Kristin M. Thompson, H. Nicholas Marsh, David M. Epstein and P. Shannon Pendergrast*

Archemix Corp. 300 3rd St. Cambridge, MA 02142, USA

*To whom correspondence should be addressed. Tel: +1 617 475 2334; Fax: +1 617 621 9300; Email: pendergrast{at}archemix.com

Received January 26, 2006. Accepted January 27, 2006.

The specific down-regulation of gene expression in cells is a powerful method for elucidating a gene's function. A common method for suppressing gene expression is the elimination of mRNA by RNAi or antisense. Alternatively, oligonucleotide-derived aptamers have been used as protein-directed agents for the specific knock-down of both intracellular and extracellular protein activity. Protein-directed methods offer the advantage of more closely mimicking small molecule therapeutics' mechanism of activity. Furthermore, protein-directed methods may synergize with RNA-directed methods since the two methods attack gene expression at different levels. Here we have knocked down a well-characterized intracellular protein's activity, NF{kappa}B, by expressing either aptamers or small interfering RNAs (siRNAs). Both methods can diminish NF{kappa}B's activity to similar levels (from 29 to 64%). Interestingly, expression of both aptamers and siRNAs simultaneously, suppressed NF{kappa}B activity better than either method alone (up to 90%). These results demonstrate that the expression of intracellular aptamers is a viable alternative to siRNA knock-down. Furthermore, for the first time, we show that the use of aptamers and siRNA together can be the most effective way to achieve maximal knock-down of protein activity.

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors


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