SignalSilence? mTOR siRNA I

• 产品编号：CST-6381S      品牌：CST       原厂货号：6381S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence® mTOR siRNA I 能够抑制人、小鼠和大鼠mTOR的表达。来自Cell Signaling Technology (CST)的SignalSilence^® mTOR siRNA I 可以帮助研究者通过RNA干扰特异性地抑制mTOR的表达，这种方法可以通过将双链RNA分子传递到细胞内从而使基因表达有选择的沉默。来自CST的所有的SignalSilence^® siRNA产品都是经过内部严格检测的，并且通过Western blot 分析证明确实能够减少目的蛋白的表达。通过三苯甲基分析每个碱基以监测寡核苷酸的合成，确保合适的配对效率。随后寡核苷酸通过亲和固相萃取法纯化。退火的RNA双链通过质谱分析来证实其精确的组成。每一批产品都通过质谱分析与前面的产品进行比较，来保证不同批次之间的最大一致性。CST推荐使用100 nM SignalSilence^® mTOR siRNA I 进行转染，48到72小时后对细胞进行裂解。转染步骤按照转染试剂说明书提供的步骤进行。遇到任何使用方面的问题，请随时联系CST。哺乳动物雷帕霉素（rapamycin）靶蛋白（mTOR，FRAP，RAFT）是一种丝氨酸/苏氨酸蛋白激酶（1-3），其作为ATP和氨基酸的传感器，可以有效调控营养供应及细胞的生长（4,5）。当营养充分时，mTOR响应磷脂酸介导的信号，进而正向调节p70 S6 激酶，并参与失活eIF4E的抑制剂4E-BP1。这些信号导致特定的mRNA亚群的翻译。mTOR可以被Akt信号通路的PI3K激酶磷酸化修饰Ser2448位点，也能自我磷酸化Ser2481位点（7,8）。mTOR的在细胞生长和动态平衡中起着关键作用，在肿瘤中可能存在调控失常。因此，目前很多研究将mTOR作为一个潜在的抗肿瘤治疗靶点（9）。siRNA已经用于在293-HEK细胞中沉默mTOR的表达，引起raptor的表达降低， p70 S6激酶磷酸化降低和细胞体积减少。

1. Sabers, C.J. et al. (1995) J Biol Chem 270, 815-22.
2. Brown, E.J. et al. (1994) Nature 369, 756-8.
3. Sabatini, D.M. et al. (1994) Cell 78, 35-43.
4. Gingras, A.C. et al. (2001) Genes Dev 15, 807-26.
5. Dennis, P.B. et al. (2001) Science 294, 1102-5.
6. Fang, Y. et al. (2001) Science 294, 1942-5.
7. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
8. Peterson, R.T. et al. (2000) J Biol Chem 275, 7416-23.
9. Huang, S. and Houghton, P.J. (2003) Curr Opin Pharmacol 3, 371-7.
10. Kim, D. H. et al. (2002) Cell 110, 163-175.

原厂资料：

Homology

Species predicted to react based on 100% sequence homology: Mouse, Rat

Specificity / Sensitivity

SignalSilence® mTOR siRNA I will inhibit human, mouse and rat mTOR expression.

Description

SignalSilence® mTOR siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit mTOR expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.

Quality Control

Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.

Directions for Use

CST recommends transfection with 100 nM mTOR siRNA I 48 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.

Background

The mammalian target of rapamycin (mTOR, FRAP, RAFT) is a Ser/Thr protein kinase (1-3) that functions as an ATP and amino acid sensor to balance nutrient availability and cell growth (4,5). When sufficient nutrients are available, mTOR responds to a phosphatidic acid-mediated signal to transmit a positive signal to p70 S6 kinase and participate in the inactivation of the eIF4E inhibitor, 4E-BP1 (6). These events result in the translation of specific mRNA subpopulations. mTOR is phosphorylated at Ser2448 via the PI3 kinase/Akt signaling pathway and autophosphorylated at Ser2481 (7,8). mTOR plays a key role in cell growth and homeostasis and may be abnormally regulated in tumors. For these reasons, mTOR is currently under investigation as a potential target for anti-cancer therapy (9).siRNA has been used to silence mTOR expression in 293-HEK cells causing decreased raptor expression, decreased p70 S6 kinase phosphorylation, and reduced cell size (9).

1. Sabers, C.J. et al. (1995) J Biol Chem 270, 815-22.
2. Brown, E.J. et al. (1994) Nature 369, 756-8.
3. Sabatini, D.M. et al. (1994) Cell 78, 35-43.
4. Gingras, A.C. et al. (2001) Genes Dev 15, 807-26.
5. Dennis, P.B. et al. (2001) Science 294, 1102-5.
6. Fang, Y. et al. (2001) Science 294, 1942-5.
7. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
8. Peterson, R.T. et al. (2000) J Biol Chem 275, 7416-23.
9. Huang, S. and Houghton, P.J. (2003) Curr Opin Pharmacol 3, 371-7.
10. Kim, D. H. et al. (2002) Cell 110, 163-175.

注意事项：

Storage: SignalSilence® siRNA is supplied in RNAse-free water. Aliquot and store at -20ºC.