TY  - JOUR
A1  - Arts, Gert-Jan
A1  - Langemeijer, Ellen
A1  - Tissingh, Rudi
A1  - Ma, Libin
A1  - Pavliska, Heidi
A1  - Dokic, Kristina
A1  - Dooijes, Richele
A1  - Mešić, Emir
A1  - Clasen, Remko
A1  - Michiels, Frits
A1  - van der Schueren, Jan
A1  - Lambrecht, Mark
A1  - Herman, Sofie
A1  - Brys, Reginald
A1  - Thys, Kim
A1  - Hoffmann, Marcel
A1  - Tomme, Peter
A1  - van Es, Helmuth
T1  - Adenoviral Vectors Expressing siRNAs for Discovery and Validation of Gene Function
Y1  - 2003/10/01 
JF  - Genome Research 
JO  - Genome Research 
SP  - 2325 
EP  - 2332 
DO  - 10.1101/gr.1332603 
VL  - 13 
IS  - 10 
UR  - http://genome.cshlp.org/content/13/10/2325.abstract 
N2  - RNA interference is a powerful tool for studying gene function and for drug target discovery in diverse organisms and cell types. In mammalian systems, small interfering RNAs (siRNAs), or DNA plasmids expressing these siRNAs, have been used to down-modulate gene expression. However, inefficient transfection protocols, in particular, for primary cell types, have hampered the use of these tools in disease-relevant cellular assays. To be able to use this technology for genome-wide function screening, a more robust transduction protocol, resulting in a longer duration of the knock-down effect, is required. Here, we describe the validation of adenoviral vectors that express hairpin RNAs that are further processed to siRNAs. Infection of cell lines, or primary human cells, with these viruses leads to an efficient, sequence-specific, and prolonged reduction of the corresponding target mRNA, resulting in a reduction of the encoded protein level in the cell. For knock-down of one of the targets, GαS, we have measured inhibition of ligand-dependant, G-protein-coupled signaling. It is expected that this technology will prove to be of great value in target validation and target discovery efforts. 
ER  -