SignalSilence? APC6 siRNA I

• 产品编号：CST-7279S      品牌：CST       原厂货号：7279S
• 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® APC6 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APC6 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。腺瘤性息肉病(APC)肿瘤抑制基因在大多数家族性和散发性结肠癌中发生突变，它编码一个大的胞质蛋白，该蛋白在细胞迁移、细胞粘附和增殖中具有重要意义(1)。APC能够直接结合微管，其缺乏可导致有丝分裂纺锤体缺陷和染色体异常分离引起的非整倍体性(2)。APC是重要的骨架蛋白，可以结合β-连环蛋白，并参与其胞内浓度的调节。在非Wnt信号通路中，GSK-3β能够磷酸化APC-β-catenin-axin复合体的三个成员，这种β-连环蛋白的磷酸化可以为泛素创建一个识别位点，也是蛋白酶体介导降解的信号。在Wnt信号通路中，散乱蛋白失活GSK-3β而β-连环蛋白协调细胞周期进程重要调节蛋白的基因转录，如周期蛋白D1和c-Myc (3)。

1. Qiao, X. et al. (2010) Cell Cycle 9, 3904-12.
2. Harper, J.W. et al. (2002) Genes Dev 16, 2179-206.
3. Chang, L. et al. (2014) Nature 513, 388-93.
4. Carroll, C.W. and Morgan, D.O. (2002) Nat Cell Biol 4, 880-7.
5. Gmachl, M. et al. (2000) Proc Natl Acad Sci U S A 97, 8973-8.
6. Leverson, J.D. et al. (2000) Mol Biol Cell 11, 2315-25.
7. Kraft, C. et al. (2005) Mol Cell 18, 543-53.
8. Glotzer, M. et al. (1991) Nature 349, 132-8.
9. Pfleger, C.M. and Kirschner, M.W. (2000) Genes Dev 14, 655-65.
10. Schreiber, A. et al. (2011) Nature 470, 227-32.
11. Kraft, C. et al. (2003) EMBO J 22, 6598-609.

原厂资料：

Description

SignalSilence® APC6 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit APC6 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® APC6 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 Adenomatous Polyposis Coli (APC) tumor suppressor gene is mutated in most familial and sporadic colorectal cancers and encodes a large cytoplasmic protein that is implicated in cell migration, cell adhesion, and proliferation (1). APC binds directly to microtubules and lack of APC leads to defective mitotic spindles and aneuploidy due to missegregation of chromosomes (2). APC is well characterized as a scaffolding protein, binds to β-catenin, and is involved in the regulation of its intracellular concentration. In the absence of a Wnt signal, GSK-3β phosphorylates all three members of the APC-β-catenin-axin complex and this phosphorylation of β-catenin creates a recognition site for ubiquitin, the signal for proteasome-mediated degradation. In the presence of a Wnt signal, dishevelled inactivates GSK-3β and β-catenin coordinates gene transcription of proteins important for the control of cell cycle progression and proliferation, such as cyclin D1 and c-Myc (3).

1. Qiao, X. et al. (2010) Cell Cycle 9, 3904-12.
2. Harper, J.W. et al. (2002) Genes Dev 16, 2179-206.
3. Chang, L. et al. (2014) Nature 513, 388-93.
4. Carroll, C.W. and Morgan, D.O. (2002) Nat Cell Biol 4, 880-7.
5. Gmachl, M. et al. (2000) Proc Natl Acad Sci U S A 97, 8973-8.
6. Leverson, J.D. et al. (2000) Mol Biol Cell 11, 2315-25.
7. Kraft, C. et al. (2005) Mol Cell 18, 543-53.
8. Glotzer, M. et al. (1991) Nature 349, 132-8.
9. Pfleger, C.M. and Kirschner, M.W. (2000) Genes Dev 14, 655-65.
10. Schreiber, A. et al. (2011) Nature 470, 227-32.
11. Kraft, C. et al. (2003) EMBO J 22, 6598-609.

注意事项：

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