SignalSilence? Cyclin D1 siRNA II (Mouse Specific)

• 产品编号：CST-6477S      品牌：CST       原厂货号：6477S
• 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® Cyclin D1 siRNA II (小鼠特异性)，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Cyclin D1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Cyclin D1 siRNA II (小鼠特异性)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。细胞周期蛋白依赖性蛋白激酶CDK4和CDK6的活性受三个因素的调控：T-环路磷酸化，周期蛋白伴侣(D-型周期蛋白)的丰度，与CDK抑制剂Cip/Kip 或 INK蛋白家族的关系(1)。失活的三元复合物cyclin D/CDK4 和p27 Kip1需要胞外有丝分裂刺激因子参与p27伴侣的释放和降解，伴随周期蛋白D的水平升高，通过限制点和pRb依赖性的S期进入影响细胞周期的进程(2)。活跃的cyclin D/CDK4复合体靶向视网膜母细胞瘤蛋白以利于磷酸化，促进可以激活G1/S期基因表达的转录因子E2F的释放(3)。生长因子退出后，通过下调蛋白表达和磷酸化依赖性蛋白降解，周期蛋白D的水平下降(4)。

1. Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681.
2. Sherr, C.J. (1996) Science 274, 1672-1677.
3. Lukas, J. et al. (1996) Mol. Cell. Biol. 16, 6917-6925.
4. Diehl, J.A. et al. (1997) Genes Dev. 11, 957-972.

原厂资料：

Description

SignalSilence® Cyclin D1 siRNA II (Mouse Specific) from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Cyclin D1 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 Cyclin D1 siRNA II (Mouse Specific) 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

Activity of the cyclin-dependent kinases CDK4 and CDK6 is regulated by T-loop phosphorylation, by the abundance of their cyclin partners (the D-type cyclins), and by association with CDK inhibitors of the Cip/Kip or INK family of proteins (1). The inactive ternary complex of cyclin D/CDK4 and p27 Kip1 requires extracellular mitogenic stimuli for the release and degradation of p27 concomitant with a rise in cyclin D levels to effect progression through the restriction point and pRb-dependent entry into S-phase (2). The active complex of cyclin D/CDK4 targets the retinoblastoma protein for phosphorylation, allowing the release of E2F transcription factors that activate G1/S-phase gene expression (3). Levels of cyclin D protein drop upon withdrawal of growth factors through downregulation of its protein expression and through phosphorylation-dependent degradation (4). 

1. Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681.
2. Sherr, C.J. (1996) Science 274, 1672-1677.
3. Lukas, J. et al. (1996) Mol. Cell. Biol. 16, 6917-6925.
4. Diehl, J.A. et al. (1997) Genes Dev. 11, 957-972.

注意事项：

Storage: Cyclin D1 siRNA II (Mouse Specific) is supplied in
RNAse-free water. Aliquot and store at -20ºC.