Published online 19 July 2006
© 2006 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-commerical use, distribution, and reproduction in any medium, provided the original work is properly cited.
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H1 RNA polymerase III promoter-driven expression of an RNA aptamer leads to high-level inhibition of intracellular protein activity
1 Department of Surgery, Duke University Medical Center Durham, NC, USA 2 Department of Psychiatry, Duke University Medical Center Durham, NC, USA 3 Department of Radiation Oncology, Duke University Medical Center Durham, NC, USA 4 Department of Medicine, Duke University Medical Center Durham, NC, USA
*To whom correspondence should be addressed at Box 3629 and Box 2633, Duke University Medical Center, Durham, NC 27710, USA. Tel: +1 919 684 3381; Fax: +1 919 668 0487; Email: clary001{at}mc.duke.edu
Received May 8, 2006. Revised June 22, 2006. Accepted June 24, 2006.
Aptamers offer advantages over other oligonucleotide-based approaches that artificially interfere with target gene function due to their ability to bind protein products of these genes with high affinity and specificity. However, RNA aptamers are limited in their ability to target intracellular proteins since even nuclease-resistant aptamers do not efficiently enter the intracellular compartments. Moreover, attempts at expressing RNA aptamers within mammalian cells through vector-based approaches have been hampered by the presence of additional flanking sequences in expressed RNA aptamers, which may alter their functional conformation. In this report, we successfully expressed a ‘pure’ RNA aptamer specific for NF-
B p50 protein (A-p50) utilizing an adenoviral vector employing the H1 RNA polymerase III promoter. Binding of the expressed aptamer to its target and subsequent inhibition of NF-B mediated intracellular events were demonstrated in human lung adenocarcinoma cells (A549), murine mammary carcinoma cells (4T1) as well as a human tumor xenograft model. This success highlights the promise of RNA aptamers to effectively target intracellular proteins for in vitro discovery and in vivo applications.
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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