Inducible and reversible suppression of Npm1 gene expression using stably integrated small interfering RNA vector in mouse embryonic stem cells
Inducible and reversible suppression of Npm1 gene expression using stably integrated small interfering RNA vector in mouse embryonic stem cells
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DOI：
10.1016/j.bbrc.2006.07.020
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发表时间：
2006-09-08
影响因子：
3.1
通讯作者：
Jin, Ying
中科院分区：
文献类型：
--
作者：
Wang, Bei Bei;Lu, Rui;Jin, Ying
四环素诱导的小干扰RNA （siRNA）是研究哺乳动物细胞基因功能的有力工具。然而，该系统很少用于胚胎干细胞（ES）。在这里，我们首次在小鼠胚胎干细胞中应用tc诱导的、稳定整合的基于质粒的siRNA系统来下调Npm1的表达，Npm1是胚胎发育的重要基因。Npm1在胚胎干细胞中的生理作用尚未明确。我们的数据表明，该siRNA系统不仅高效地敲除Npm1的表达，而且与tc剂量和诱导时间相关。特别是Npm1表达的下调是可逆的。重要的是，抑制胚胎干细胞中Npm1的表达导致细胞增殖减少。综上所述。该系统允许以高度可控的方式研究基因功能，否则难以在胚胎干细胞中实现。此外，我们的研究结果表明，Npm1对胚胎干细胞增殖至关重要。(c) 2006爱思唯尔公司版权所有。
The tetracycline (Tc)-inducible small interference RNA (siRNA) is a powerful toot 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. (c) 2006 Elsevier Inc. All rights reserved.