SignalSilence? MUC1 siRNA I

• 产品编号：CST-13253S      品牌：CST       原厂货号：13253S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

Cell Signaling Technology (CST) 公司的SignalSilence® MUC1 siRNA I 帮助研究人员使用RNA干扰技术特异性抑制MUC1表达，该技术通过向细胞内导入双链RNA分子以选择性的抑制某些基因的表达。所有CST公司的SignalSilence® siRNA都经过了严格的内部测试，western blot分析显示能够降低目标蛋白的表达。寡核苷酸合成经过了三苯甲基的逐碱基分析以确保耦合效率。随后经过亲和固相萃取纯化。退火的RNA双链在经过质谱分析以确保双链的准确性。新批次都会经过质谱分析与之前的批次进行比对以保证不同批次之间的稳定性。CST建议转染100 nM SignalSilence® MUC1 siRNA I，48-72小时后裂解细胞。转染的步骤请参阅转染试剂的说明书。关于使用问题，可以随时咨询CST。 每管试剂能进行100次转染，该转染次数以总体积为300μl 的1个24孔板孔中siRNA终浓度100 nM来进行计算。Mucins代表了一个家族的糖蛋白，它们的特征是重复的结构域和高度的O-糖基化(1).MUC1(或mucin 1)在大多数人类癌症中异常高表达。肿瘤中MUC1表达升高会减少细胞-细胞和细胞-细胞外基质之间的相互作用。MUC1可以被蛋白水解，大的胞外结构域可以与含有跨膜片段和72个残基的胞质尾区小的25kDa的羧基端结构域保持相互关联(1)。MUC1和ErbB家族受体相互作用激活ERK1/2（2）。MUC1也可以和β-catenin相互作用，GSK-3β, PKCγ和Src可以通过磷酸化MUC1胞质尾区的Ser44, Thr41和Tyr46调控MUC1和β-catenin的相互作用(3-5)。过表达MUC1会增加细胞转化(6)，并通过Akt蛋白或p53信号通路衰减压力诱导的凋亡(7,8)。

1. Baldus, S.E. et al. (2004) Crit Rev Clin Lab Sci 41, 189-231.
2. Schroeder, J.A. et al. (2001) J Biol Chem 276, 13057-64.
3. Li, Y. et al. (1998) Mol Cell Biol 18, 7216-24.
4. Li, Y. et al. (2001) J Biol Chem 276, 6061-4.
5. Ren, J. et al. (2002) J Biol Chem 277, 17616-22.
6. Schroeder, J.A. et al. (2004) Oncogene 23, 5739-47.
7. Raina, D. et al. (2004) J Biol Chem 279, 20607-12.
8. Wei, X. et al. (2005) Cancer Cell 7, 167-78.

原厂资料：

Description

SignalSilence^® MUC1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit MUC1 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^® MUC1 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.

Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

Background

Mucins represent a family of glycoproteins characterized by repeat domains and dense O-glycosylation (1). MUC1 (or mucin 1) is aberrantly overexpressed in most human carcinomas. Increased expression of MUC1 in carcinomas reduces cell-cell and cell-ECM interactions. MUC1 is cleaved proteolytically, and the large ectodomain can remain associated with the small 25 kDa carboxy-terminal domain that contains a transmembrane segment and a cytoplasmic tail of 72 residues (1). MUC1 interacts with ErbB family receptors and potentiates ERK1/2 activation (2). MUC1 also interacts with β-catenin, which is regulated by GSK-3β, PKCγ and Src through phosphorylation at Ser44, Thr41 and Tyr46 of MUC1 cytoplasmic tail (3-5). Overexpression of MUC1 potentiates transformation (6), and attenuates stress-induced apoptosis through the Akt or p53 pathways (7,8).

1. Baldus, S.E. et al. (2004) Crit Rev Clin Lab Sci 41, 189-231.
2. Schroeder, J.A. et al. (2001) J Biol Chem 276, 13057-64.
3. Li, Y. et al. (1998) Mol Cell Biol 18, 7216-24.
4. Li, Y. et al. (2001) J Biol Chem 276, 6061-4.
5. Ren, J. et al. (2002) J Biol Chem 277, 17616-22.
6. Schroeder, J.A. et al. (2004) Oncogene 23, 5739-47.
7. Raina, D. et al. (2004) J Biol Chem 279, 20607-12.
8. Wei, X. et al. (2005) Cancer Cell 7, 167-78.

注意事项：

Storage: MUC1 siRNA is supplied in RNAse-free water. Aliquot
and store at -20ºC.