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Mol Biotechnol (2008) 40:1926 DOI 10.1007/s12033-008-9043-x
RESEARCH
Lipofectamine RNAiMAX: An Efcient siRNA Transfection Reagent in Human Embryonic Stem Cells
Ming Zhao Hong Yang Xingjun Jiang Wen Zhou Bin Zhu Ying Zeng Kaitai Yao Caiping Ren
Published online: 8 March 2008 Humana Press 2008
Abstract RNA interference methodology suppresses specic gene expression, thus mimicking loss-of-function mutation and enabling in vitro and in vivo gene function analysis. Lipofectamine RNAiMAX, a new transfection reagent, has been conrmed high efciency in delivering small interfering RNA (siRNA) into mesenchymal stem cells and neural stem cells. In this study, we used three transfection reagents (Lipofectamine RNAiMAX, Oligofectamine and Lipofectamine 2000) to deliver siRNA into human embryonic stem (hES) cells and compared the silencing efciency of enhanced green uorescent protein transgene and Oct4, a key regulator of pluripotency. As a result, siRNA can be delivered into hES cells more efciently by Lipofectamine RNAiMAX compared with the other two transfection reagents and high efcient knockdown of target genes was obtained by Lipofectamine RNAiMAX even at a low concentration of siRNA. Quantitative real-time PCR showed approximately 90% knockdown of Oct4 transcript with cognate Oct4 siRNA transfection by Lipofectamine RNAiMAX compared to control siRNA. These results demonstrated that Lipofectamine RNAiMAX is an efcient and excellent transfection reagent for delivering siRNA into hES cells.
Keywords Human embryonic stem cells
Lipofectamine RNAiMAX Oct4 siRNA Transfection
Introduction
Human embryonic stem (hES) cells offer the opportunity for in vitro production of multiple cell types for use in regenerative medicine [1]. The self-renewal and differentiation of hES cells is under the control of many genes and signaling pathways, however, the mechanism has still not been demonstrated completely. The technologies for transgene expression and gene loss of function have been developed for studying gene function in cells, which provides a chance for elucidating the roles of genes in hES cells. In recent years, small interfering RNA (siRNA) duplexes and RNA interference (RNAi) vectors have been successfully applied to suppressing multiple genes in mammalian cells including hES cells [24]. Compared to RNAi vectors, siRNA duplexes provide a faster and more convenient method to knock down gene expression. However, to date RNAi has not been exploited with high efcacy in hES cells. A leading cause of inefcient RNAi knockdown is the poor transfection...