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Nucleic Acids Research 2005 33(8):e74; doi:10.1093/nar/gni068
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Published online 28 April 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

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Suppression of vascular endothelial growth factor expression at the transcriptional and post-transcriptional levels

Heung-Sun Kwon, Hyun-Chul Shin and Jin-Soo Kim*

ToolGen, Inc. 461-6, Jeonmin-dong, Yuseong-gu, Daejeon 305-390, South Korea

*To whom correspondence should be addressed. Tel: +82 42 863 8166; Fax: +82 42 863 3840; Email: jsk{at}toolgen.com

Received January 10, 2005. Revised March 30, 2005. Accepted March 30, 2005.

Gene expression is regulated at the transcriptional and post-transcriptional levels. Therefore, in order to achieve a high level of silencing, which includes minimizing any residual expression of a target gene, suppression at both the transcriptional and post-transcriptional levels is required. In this study, we describe a new method for highly efficient gene silencing that combines zinc finger protein-mediated transcriptional repression and small interfering RNA (siRNA)-mediated inhibition of post-transcriptional events. To measure the amount of gene expression under various conditions, we used a luciferase reporter gene that was driven by a variety of promoters, including that of the human vascular endothelial growth factor-A (VEGF-A) gene. We also measured expression of the endogenous VEGF-A gene. Inhibition of gene expression by each of the two individual technologies was effective, but in-depth analyses revealed residual expression of the target gene. The combination of specific zinc finger transcription factors and siRNAs greatly enhanced the silencing of the human VEGF-A gene, not only when cells were grown in the presence of normal amounts of oxygen but also under conditions of hypoxic stimulation. These results suggest that a bi-level approach to the silencing of VEGF-A expression may be clinically beneficial as part of a cancer treatment protocol.

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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