SignalSilence? DDX5 siRNA I

• 产品编号：CST-8626S      品牌：CST       原厂货号：8626S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence® DDX5 siRNA I将抑制人、小鼠、大鼠和猴DDX5蛋白表达。Cell Signaling Technology (CST)公司的SignalSilence® DDX5 siRNA I允许研究者通过RNA干扰去特异性抑制DDX5蛋白表达，而RNA干扰是通过双链RNA分子传送到细胞中能选择性沉默基因表达的一个方法。所有来自CST公司的SignalSilence® siRNA产品都经过严格的内部检测，通过western blot分析显示可降低靶蛋白表达。通过三苯基分析对寡核苷酸合成进行逐个碱基的监测以保证适当的耦合效率，随后通过亲和固相提取来纯化寡核苷酸。退火的双链RNA需要通过质谱进行进一步的分析从而确定提取物的组成。为了保证批次间一致性最大化，每一批次产品都要通过质谱与前一批次的产品进行比较。CST建议使用100 nM SignalSilence® AUF1/hnRNP D siRNA I进行转染，并在48-72小时后裂解细胞。具体的转染操作步骤请根据转染试剂生产商提供的方法进行。如您在使用过程中有任何问题请随时联系我们。每瓶中含有100次转染所需用量。每次转染siRNA的终浓度为100 nM，以上用量为24孔板中转染，每孔总体积为300 μl。DDX5 (DEAD盒子多肽5)又称为p68，其起初被鉴定为和翻译起始因子eIF-4A蛋白类似的一个68 kDa大小的核蛋白(1)。DDX5是推测的DEAD盒子RNA解螺酶家族成员，通常其含有保守的DEAD (Asp-Glu-Ala-Asp)基序，而该基序的主要功能是调节RNA的二级结构。DDX5蛋白具有ATP-依赖的RNA解旋酶活性，以及该蛋白作为调节miRNA和rRNA加工的的DROSHA复合物重要组分(3,4)。DDX也可能调节mRNA剪接(5)，并且证实和HDAC1相互作用，而HDAC1是能够调节启动子特异性转录(6)。DDX5能够和多种不同的蛋白质相互作用，包括Runx2, p53, Smad3, CBP和p300 (7-10)，研究表明DDX5蛋白在生物发育进程中的多个维度起到重要作用。尤其是DDX5蛋白可能涉及到生长因子诱导上皮-间质转化（EMT）。在PDGF刺激下DDX5在Tyr593位点磷酸化，使Axin与β-catenin解离； 这阻断了GSK-3β对β-catenin磷酸化，从而促进Wnt蛋白依赖的β-catenin核迁移(11)以及促使c-Myc, cyclin D1和Snai1基因转录(12,13)。

1. Ford, M.J. et al. (1988) Nature 332, 736-8.
2. Hirling, H. et al. (1989) Nature 339, 562-4.
3. Fukuda, T. et al. (2007) Nat Cell Biol 9, 604-11.
4. Davis, B.N. et al. (2008) Nature 454, 56-61.
5. Camats, M. et al. (2008) PLoS ONE 3, e2926.
6. Wilson, B.J. et al. (2004) BMC Mol Biol 5, 11.
7. Jensen, E.D. et al. (2008) J Cell Biochem 103, 1438-51.
8. Bates, G.J. et al. (2005) EMBO J 24, 543-53.
9. Warner, D.R. et al. (2004) Biochem Biophys Res Commun 324, 70-6.
10. Rossow, K.L. and Janknecht, R. (2003) Oncogene 22, 151-6.
11. Yang, L. et al. (2006) Cell 127, 139-55.
12. Yang, L. et al. (2007) J Biol Chem 282, 16811-9.
13. Carter, C.L. et al. (2010) Oncogene 29, 5427-36.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence® DDX5 siRNA I will inhibit human, mouse, rat, and monkey DDX5 expression.

Description

SignalSilence® DDX5 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit DDX5 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® DDX5 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

DDX5 (DEAD box polypeptide 5), also known as p68, was first identified as a 68 kDa nuclear protein with similarity to translation initiation factor eIF-4A (1). DDX5 is a member of the DEAD box family of putative RNA helicases, defined by the presence of a conserved DEAD (Asp-Glu-Ala-Asp) motif that appears to function primarily in the regulation of RNA secondary structure. DDX5 exhibits ATP-dependent RNA helicase activity (2) and has been identified as a critical subunit of the DROSHA complex that regulates miRNA and rRNA processing (3,4). DDX may also regulate mRNA splicing (5) and has been shown to interact with HDAC1, where it can regulate promoter-specific transcription (6). DDX5 interacts with a diverse group of proteins, including Runx2, p53, Smad3, CBP, and p300 (7-10), suggesting an important role for DDX5 in a multitude of developmental processes. Notably, DDX5 may be involved in growth factor-induced epithelial mesechymal transition (EMT). Phosphorylation of DDX5 at Tyr593 following PDGF stimulation was shown to displace Axin from β-catenin; this prevented phosphorylation of β-catenin by GSK-3β, leading to Wnt-independent nuclear translocation of β-catenin (11) and increased transcription of c-Myc, cyclin D1, and Snai1 (12,13).

1. Ford, M.J. et al. (1988) Nature 332, 736-8.
2. Hirling, H. et al. (1989) Nature 339, 562-4.
3. Fukuda, T. et al. (2007) Nat Cell Biol 9, 604-11.
4. Davis, B.N. et al. (2008) Nature 454, 56-61.
5. Camats, M. et al. (2008) PLoS ONE 3, e2926.
6. Wilson, B.J. et al. (2004) BMC Mol Biol 5, 11.
7. Jensen, E.D. et al. (2008) J Cell Biochem 103, 1438-51.
8. Bates, G.J. et al. (2005) EMBO J 24, 543-53.
9. Warner, D.R. et al. (2004) Biochem Biophys Res Commun 324, 70-6.
10. Rossow, K.L. and Janknecht, R. (2003) Oncogene 22, 151-6.
11. Yang, L. et al. (2006) Cell 127, 139-55.
12. Yang, L. et al. (2007) J Biol Chem 282, 16811-9.
13. Carter, C.L. et al. (2010) Oncogene 29, 5427-36.

注意事项：

Storage: DDX5 siRNA I is supplied in RNAse-free water. Aliquot
and store at -20ºC.