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          Institute: MPI für molekulare Zellbiologie und Genetik     Collection: Publikationen MPI-CBG 2004     Display Documents



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ID: 229774.0, MPI für molekulare Zellbiologie und Genetik / Publikationen MPI-CBG 2004
Generation of single and double knockdowns in polarized epithelial cells by retrovirus-mediated RNA interference.
Authors:Schuck, Sebastian; Manninen, Aki; Honsho, Masanori; Fullekrug, Joachim; Simons, Kai
Date of Publication (YYYY-MM-DD):2004
Title of Journal:Proceedings of the National Academy of Sciences, USA
Volume:101
Issue / Number:14
Start Page:4912
End Page:4917
Copyright:not available
Review Status:not specified
Audience:Experts Only
Intended Educational Use:No
Abstract / Description:RNA interference (RNAi) is a ubiquitous mechanism of eukaryotic gene regulation that can be exploited for specific gene silencing. Retroviruses have been successfully used for stable expression of short hairpin RNAs in mammalian cells, leading to persistent inhibition of gene expression by RNAi. Here, we apply retrovirus-mediated RNAi to epithelial Madin-Darby canine kidney cells, whose properties limit the applicability of other RNAi methods. We demonstrate efficient suppression of a set of 13 target genes by retroviral coexpression of short hairpin RNAs and a selectable marker. We characterize the resulting knockdown cell populations with regard to composition and stability, and examine the usefulness of proposed guidelines for choosing RNAi target sequences. Finally, we show that this system can be used to simultaneously target two genes, giving rise to double knockdowns. Thus, retrovirus-mediated RNAi is a convenient method for gene silencing in Madin-Darby canine kidney cells, and is likely to be applicable to virtually any mammalian cell.
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für molekulare Zellbiologie und Genetik
Identifiers:LOCALID:369
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