Inducible and reversible suppression of Npm1 gene expression using stably integrated small interfering RNA vector in mouse embryonic stem cells.
The tetracycline (Tc)-inducible small interference RNA (siRNA) is a powerful tool for studying gene function in mammalian cells. However, the system is infrequently utilized in embryonic stem (ES) cells. Here, we present the first application of the Tc-inducible, stably integrated plasmid-based siRNA system in mouse ES cells to down-regulate expression of Npm1, an essential gene for embryonic development. The physiological role of Npm1 in ES cells has not been defined. Our data show that the knock-down of Npm1 expression by this siRNA system was not only highly efficient, but also Tc- dose- and induction time-dependent. Particularly, the down-regulation of Npm1 expression was reversible. Importantly, suppression of Npm1 expression in ES cells resulted in reduced cell proliferation. Taken together, this system allows for studying gene function in a highly controlled manner, otherwise difficult to achieve in ES cells. Moreover, our results demonstrate that Npm1 is essential for ES cell proliferation.
Wang
Bei Bei
BB
Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine, 225 South Chongqing Road, Shanghai 200025, China.
Lu
Rui
R
Wang
Wei Cheng
WC
Jin
Ying
Y
D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D005822:Genetic Vectors; D051379:Mice; D009687:Nuclear Proteins / Q000235:genetics* / Q000502:physiology; D000090243:Nucleophosmin; D034741:RNA, Small Interfering / Q000235:genetics*; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism*; D013752:Tetracycline / Q000494:pharmacology
C000715570:Npm1 protein, mouse; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D000090243:Nucleophosmin; D013752:Tetracycline