SignalSilence? CDK8 siRNA I

• 产品编号：CST-6438S      品牌：CST       原厂货号：6438S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

研究人员可以使用CST的SignalSilence® CDK8 siRNA I，采用RNA干扰（一种通过传递外源性的双链RNA分子到细胞中从而能选择性的沉默基因表达的方法）技术特异性的抑制CDK8蛋白表达。CST所有的SignalSilence® siRNA产品都经过了严格的内部测试并采用Western分析证明其可以有效降低靶蛋白的表达。为了保证适当的结合效率，我们采用三苯甲基分析的方法逐个碱基的监测寡核苷酸的合成。随后通过亲和固相提取纯化寡核苷酸。退火的双链RNA进一步通过质谱分析来验证双链复合体的确切成分。每一批产品和前一批都要通过质谱分析进行比较以确定批次间最大限度的一致性。CST建议使用SignalSilence® CDK8 siRNA I转染48-72小时后裂解细胞。转染的具体步骤请参考转染试剂制造商提供的操作标准。使用过程中有任何问题请随时联系CST。哺乳动物中介体复合物是一个多亚基的蛋白复合物，它能够偶联特异性的转录调节子至RNA聚合酶II (Pol II)和基础的转录子装置。不同的中介体亚基和转录因子的相互作用允许特异性的基因调节(回顾1)。中介体复合物相互作用控制各种生物学过程，包括胰岛素信号(2)，NF-κB-依赖的信号(3)，干细胞多能性和自我更新(4,5)和结肠癌细胞增殖(6,7)。CDK8/Cyclin C，连同Med12和Med13，在中介体复合物内部组成亚复合体，可以充当分子开关的角色，抑制Pol II招募和转录起始(8,9)。CDK8的表达能够废除结肠癌细胞中E2F-1-依赖的β-连环蛋白的活性(9)。高水平的CDK8与结肠癌细胞中高水平的β-连环蛋白依赖的转录相一致，而且其增殖可以通过抑制CDK8的表达被抑制(8)。

1. Malik, S. and Roeder, R.G. (2005) Trends Biochem Sci 30, 256-63.
2. Wang, W. et al. (2009) Dev Cell 16, 764-71.
3. van Essen, D. et al. (2009) PLoS Biol 7, e73.
4. Tutter, A.V. et al. (2009) J Biol Chem 284, 3709-18.
5. Varelas, X. et al. (2008) Nat Cell Biol 10, 837-48.
6. Firestein, R. et al. (2008) Nature 455, 547-51.
7. Morris, E.J. et al. (2008) Nature 455, 552-6.
8. Knuesel, M.T. et al. (2009) Mol Cell Biol 29, 650-61.
9. Knuesel, M.T. et al. (2009) Genes Dev 23, 439-51.

原厂资料：

Homology

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

Specificity / Sensitivity

CDK8 siRNA I inhibits human, mouse, and rat CDK8 expression. 

Description

SignalSilence® CDK8 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit CDK8 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 SignalSilence® CDK8 siRNA I 48 to 72 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 Mediator Complex is a multi-subunit protein complex that couples specific transcriptional regulators to RNA polymerase II (Pol II) and the basal transcription machinery. Interactions between distinct Mediator subunits and transcription factors allow for specific gene regulation (reviewed in 1). 
 Mediator complex interactions control various biological processes including insulin signaling (2), NF-κB-dependent signaling (3), stem cell pluripotency and self renewal (4,5), and proliferation of colon cancer cells (6,7). 
 CDK8/Cyclin C, along with Med12 and Med13, constitutes a subcomplex within the Mediator Complex thought to act as a molecular switch, inhibiting Pol II recruitment and transcription initiation (8,9). Expression of CDK8 abrogates E2F-1-dependent inhibition of β-catenin activity in colon cancer cells (9). High levels of CDK8 coincide with high β-catenin-dependent transcription in colon cancer cells, and their proliferation can be inhibited by suppressing CDK8 expression (8). 

1. Malik, S. and Roeder, R.G. (2005) Trends Biochem Sci 30, 256-63.
2. Wang, W. et al. (2009) Dev Cell 16, 764-71.
3. van Essen, D. et al. (2009) PLoS Biol 7, e73.
4. Tutter, A.V. et al. (2009) J Biol Chem 284, 3709-18.
5. Varelas, X. et al. (2008) Nat Cell Biol 10, 837-48.
6. Firestein, R. et al. (2008) Nature 455, 547-51.
7. Morris, E.J. et al. (2008) Nature 455, 552-6.
8. Knuesel, M.T. et al. (2009) Mol Cell Biol 29, 650-61.
9. Knuesel, M.T. et al. (2009) Genes Dev 23, 439-51.

注意事项：

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