Adipocyte differentiation induced using nonspecific siRNA controls in cultured human mesenchymal stem cells

Xu, Yunhe, Mirmalek-Sani, Sayed-Hadi, Lin, Feng, Zhang, Junlong and Oreffo, Richard O.C. (2007) Adipocyte differentiation induced using nonspecific siRNA controls in cultured human mesenchymal stem cells. RNA, 13 (8), 1179-1183. (doi:10.1261/rna.527207).

Abstract

RNA interference (RNAi) is gene silencing induced by double-stranded RNA of 21-23 nucleotides in length, termed small interfering RNA, or siRNA. RNAi-based techniques have been widely applied to elucidate gene function, identify drug targets, and used in trials as a promising adjunct to silence disease-causing genes. However, emerging evidence suggests unexpected changes in expression of untargeted genes as a consequence of an off-target effect by RNAi in mammalian cells. To date, our understanding of such effects on stem cells is limited. We transfected human fetal femur-derived mesenchymal stem cells using commercially available nonspecific siRNA controls and examined adipocyte differentiation in the cells using morphology, histochemistry, and quantitative real-time PCR to examine the expression of key genes for adipogenic or osteogenic differentiation. We report here the induction of adipocyte differentiation in human mesenchymal stem cells using nonspecific siRNAs raising concerns as to the specificity of RNAi in stem cells and, critically, a need to understand and delineate the rules governing the specificity of RNAi

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Contributors

Author: Richard O.C. Oreffo

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