Silencing of TonEBP/NFAT5 transcriptional activator by RNA interference
Silencing of TonEBP/NFAT5 transcriptional activator by RNA interference
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DOI：
10.1097/01.asn.0000045050.19544.b2
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发表时间：
2003-02-01
影响因子：
13.6
通讯作者：
Kwon, HM
中科院分区：
文献类型：
--
作者：
Na, KY;Woo, SK;Kwon, HM
TonEBP是一种转录激活因子，在许多组织和细胞类型的整个发育过程中表达。在肾髓质，TonEBP似乎是一个重要的局部调节分化凭借刺激几个基因。为了研究TonEBP的功能，开发了两种小干扰RNA（siRNA）双链体，其通过RNA干扰有效地降低TonEBP的表达。在引入TonEBP-siRNA后沉默仅持续3天。正如预期的那样，TonEBP驱动的报告基因表达和钠/肌醇共转运蛋白（SMIT），醛糖还原酶（AR）和热休克蛋白70（HSP 70）mRNA的表达显着降低细胞中TonEBP表达沉默。这些数据提供了直接的证据，SMIT，AR和HSP 70基因是TonEBP的靶点，尽管不能排除其他蛋白质（如辅助蛋白）的潜在作用。TonEBP-siRNA是一种有效的工具，应该证明在细胞中功能丧失关系的研究中是有用的。
TonEBP is a transcriptional activator that is expressed throughout development in many tissues and cell types. In the kidney medulla, TonEBP appears to be an important local regulator of differentiation by virtue of stimulating several genes. To study the function of TonEBP, two small interfering RNA (siRNA) duplexes were developed that reduced TonEBP expression effectively via RNA interference. The silencing lasted only 3 d after introduction of the TonEBP-siRNA's. As expected, TonEBP-driven reporter gene expression and expression of the sodium/myo-inositol cotransproter (SMIT), aldose reductase (AR) and heat shock protein 70 (HSP70) mRNA were significantly decreased in cells where TonEBP expression was silenced. These data provide direct evidence that the SMIT, AR, and HSP70 genes are targets of TonEBP, although the potential role of other proteins, such as accessory proteins, cannot be excluded. The TonEBP-siRNA is an effective tool that should prove useful in the investigation of loss-of-function relationship in cells.