TAILIEUCHUNG - Báo cáo khoa học: 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

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 easi-est approach to loss-of-function experiments. | ễFEBS Journal 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 Sébastien Vankoningsloo1 Franọoise de Longueville2 Stephanie Evrard2 Pierre Rahier2 Andree l-l A11 hir n1 A ntni no Ca Ýannií-ili1 álania r ae olliot 1 l - có Romanic2 Martino Raoc1 Pa rinia Ronard1 nou uion Hl noil ie rauauuioii Ivieianie xjasie I Iiei u ose neiiiauie Iviaiune naes rauiuia rienaivi and Thierry Arnould1 1 Laboratoire de Biochimie et Biologie Cellulaire University of Namur Belgium 2 Eppendorf Array Technologies Namur Belgium Keywords cell type gene expression off-target effects silencing siRNA Correspondence T. Arnould Laboratoire de Biochimie et Biologie Cellulaire University of Namur 61 rue de Bruxelles 5000 Namur Belgium Fax 32 81 724125 Tel 32 81 724129 E-mail Received 10 January 2008 revised 12 March 2008 accepted 19 March 2008 doi 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 .

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