SignalSilence? Frizzled6 siRNA II

• 产品编号：CST-6597S      品牌：CST       原厂货号：6597S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

来自Cell Signaling Technology (CST)的SignalSilence^® Frizzled6 siRNA II可以帮助研究者通过RNA干扰特异性地抑制Frizzled6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^® Frizzled6 siRNA II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。Frizzled (Fzd)属于七次跨膜G-蛋白偶联受体超家族（1）。Fzds拥有一个巨大的细胞外N末端区域，该区域拥有一个富含半胱氨酸的结构域（CRD），该结构域与Wnt蛋白的结合有关（1,2）。Fzd下游的主要信号时通过细胞内的C-末端与Dvl蛋白的PDZ结构域结合而传递的（3）。Wnt蛋白与Fzd和共同受体LRP5或LRP6结合，通过抑制GSK3-β对β-catenin的磷酸化激活Wnt/β-catenin信号通路（4,5）。除了经典的Wnt/β-catenin信号通路以外，一些Wnt蛋白也可以激活Fzd/Ca2+信号通路和Fzd/PCP（平面细胞极性）信号通路（6,7）。哺乳动物Fzd超家族有10各成员（Fzd1到Fzd10），他们可能介导了不同的信号通路（8）。一些Fzd也可以与其他的分泌型蛋白，如Norrin和R-Spondin结合（9-11）。

1. Schulte, G. and Bryja, V. (2007) Trends Pharmacol Sci 28, 518-25.
2. Hsieh, J.C. et al. (1999) Proc Natl Acad Sci USA 96, 3546-51.
3. Wong, H.C. et al. (2003) Mol Cell 12, 1251-60.
4. Zeng, X. et al. (2005) Nature 438, 873-7.
5. Davidson, G. et al. (2005) Nature 438, 867-72.
6. Fanto, M. and McNeill, H. (2004) J Cell Sci 117, 527-33.
7. Kohn, A.D. and Moon, R.T. Cell Calcium 38, 439-46.
8. Cadigan, K.M. and Liu, Y.I. (2006) J Cell Sci 119, 395-402.
9. Xu, Q. et al. (2004) Cell 116, 883-95.
10. Nam, J.S. et al. (2006) J Biol Chem 281, 13247-57.
11. Hendrickx, M. and Leyns, L. (2008) Dev Growth Differ 50, 229-43.

原厂资料：

Description

SignalSilence® Frizzled6 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Frizzled6 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 Frizzled6 siRNA II 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

Frizzled (Fzd) belongs to the seven transmembrane-spanning G-protein-coupled receptor (GPCR) superfamily (1). Fzds have a large extracellular N-terminal region containing a cysteine-rich domain (CRD), which is involved in binding to Wnt proteins (1,2). The intracellular C-terminus binds to the PDZ domain of Dvl proteins, a major signaling component downstream of Fzd (3). Wnt proteins bind to Fzd and the co-receptors LRP5 or LPR6, and activate Wnt/β-catenin pathway through inhibiting phosphorylation of β-catenin by GSK3-β (4,5). In addition to this canonical Wnt/β-catenin pathway, some Wnt proteins can also activate the Fzd/Ca2+ pathway and Fzd/PCP (planar cell polarity) pathway (6,7). The mammalian Fzd subfamily has 10 members (Fzd1 to Fzd10) and they may mediate signaling through different pathways (8). Some Fzds can also bind to other secreted proteins, like Norrin and R-Spondin (9-11).

1. Schulte, G. and Bryja, V. (2007) Trends Pharmacol Sci 28, 518-25.
2. Hsieh, J.C. et al. (1999) Proc Natl Acad Sci USA 96, 3546-51.
3. Wong, H.C. et al. (2003) Mol Cell 12, 1251-60.
4. Zeng, X. et al. (2005) Nature 438, 873-7.
5. Davidson, G. et al. (2005) Nature 438, 867-72.
6. Fanto, M. and McNeill, H. (2004) J Cell Sci 117, 527-33.
7. Kohn, A.D. and Moon, R.T. Cell Calcium 38, 439-46.
8. Cadigan, K.M. and Liu, Y.I. (2006) J Cell Sci 119, 395-402.
9. Xu, Q. et al. (2004) Cell 116, 883-95.
10. Nam, J.S. et al. (2006) J Biol Chem 281, 13247-57.
11. Hendrickx, M. and Leyns, L. (2008) Dev Growth Differ 50, 229-43.

注意事项：

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