IngentaConnect Gene expression silencing with specific small interfering RNA goe...

Authors: Vankoningsloo, Sébastien¹; de Longueville, Françoise²; Evrard, Stéphanie²; Rahier, Pierre²; Houbion, Andrée¹; Fattaccioli, Antoine¹; Gastellier, Mélanie¹; Remacle, José²; Raes, Martine¹; Renard, Patricia¹; Arnould, Thierry¹

Source: FEBS Journal, Volume 275, Number 11, June 2008 , pp. 2738-2753(16)

Publisher: Blackwell Publishing

Abstract:

RNA-mediated gene silencing (RNA interference) is a powerful way to knock down gene expression and has revolutionized the fields of cellular and molecular biology. Indeed, the transfection of cultured cells with small interfering RNAs (siRNAs) is currently considered to be the best and easiest approach to loss-of-function experiments. However, several recent studies underscore the off-target and potential cytotoxic effects of siRNAs, which can lead to the silencing of unintended mRNAs. In this study, we used a low-density microarray to assess gene expression modifications in response to five different siRNAs in various cell types and transfection conditions. We found major differences in off-target signature according to: (a) siRNA sequence; (b) cell type; (c) duration of transfection; and (d) post-transfection time before analysis. These results contribute to a better understanding of important parameters that could impact on siRNA side effects in knockdown experiments.

Keywords: cell type; gene expression; off-target effects; silencing; siRNA

Document Type: Research article

DOI: 10.1111/j.1742-4658.2008.06415.x

Affiliations: 1: Laboratoire de Biochimie et Biologie Cellulaire, University of Namur (F.U.N.D.P), Belgium 2: Eppendorf Array Technologies, Namur, Belgium

You have access to the full text article on a website external to Ingentaconnect.

Please click here to view this article on InterScience.

You may be required to register and activate access on InterScience before you can obtain the full text. If you have any queries please contact onlinehelp@oxon.blackwellpublishing.com

Gene expression silencing with `specific' small interfering RNA goes beyond specificity - a study of key parameters to take into account in the onset of small interfering RNA off-target effects