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Nucleic Acids Research Advance Access originally published online on December 7, 2006
Nucleic Acids Research 2007 35(2):e10; doi:10.1093/nar/gkl1045
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Nucleic Acids Research, 2007, Vol. 35, No. 2 e10
© 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-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

Development of a species-specific RNA polymerase I-based shRNA expression vector

M. S. Brenz Verca1,*, Peter Weber1, Christine Mayer2, Cornelia Graf1, Damián Refojo1, Ralf Kühn3, Ingrid Grummt2 and Beat Lutz1,4,*

1 Max Planck Institute of Psychiatry Kraepelinstrasse 2-10, D-80804 Munich, Germany 2 Division of Molecular Biology of the Cell II, German Cancer Research Center Im Neuenheimer Feld 581, D-69120 Heidelberg, Germany 3 Institute for Developmental Genetics/GSF Ingolstädter Landstrasse 1, D-85764 Neuherberg, Germany 4 Department of Physiological Chemistry, Johannes Gutenberg-University Mainz Duesbergweg 6, D-55099 Mainz, Germany

*To whom correspondence should be addressed. Tel: +49 6131 3925912; Fax: +49 6131 3923536; Email: blutz{at}uni-mainz.de

Received December 22, 2005. Revised November 3, 2006. Accepted November 5, 2006.

RNA interference (RNAi) can be induced in vitro either by application of synthetic short interfering RNAs (siRNAs), or by intracellular expression of siRNAs or short hairpin RNAs (shRNAs) from transfected vectors. The most widely used promoters for siRNA/shRNA expression are based on polymerase III (Pol III)-dependent transcription. We developed an alternative vector for siRNA/shRNA expression, using a mouse RNA polymerase I (Pol I) promoter. Pol I-dependent transcription serves in cells for production of ribosomal RNA (rRNA), and as such, is ubiquitously and stably active in different cell types. As Pol I-dependent transcription is highly species-specific, Pol I-based system provides an important biosafety advantage with respect to silencing of genes with unknown functions.

*Correspondence may also be addressed to M. S. Brenz Verca. Tel: +49 89 30 622 589; Fax: +49 89 30 622 610; Email: mbrenz{at}mpipsykl.mpg.de

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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