SignalSilence? Smad1 siRNA I

• 产品编号：CST-6223S      品牌：CST       原厂货号：6223S
• 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^® Smad1 siRNA I可以帮助研究者通过RNA干扰特异性地抑制Smad1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^® Smad1 siRNA I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。骨形成蛋白由一大类信号分子组成，这些分子调控包括形态发生、细胞命运定向、增殖、分化以及凋亡在内的广泛重要过程[1,2]。BMP 受体为TGF-β家族丝氨酸/苏氨酸受体成员。配基的结合诱导多聚化、自磷酸化和相应受体的激活[3-5]。它们随后磷酸化Smad1蛋白的C-末端SSXS模体中的463位丝氨酸和465位丝氨酸，且磷酸化Smad5和Smad8中相应的位置。这些磷酸化的Smad与共激活的Smad4形成二聚体，转移到核内激活靶基因的转录[5]。促分裂原活化蛋白激酶MAPK和周期蛋白依赖性蛋白激酶8和9在Smad1的连接区域磷酸化相应残基包括丝氨酸206。进而募集Smurf1到接头区域导致Smad1的降解[6]。这一位点的磷酸化也招募YAP到接头区域促进Smad1的转录激活[7]。

1. Hogan, B.L. (1996) Genes Dev 10, 1580-94.
2. Hoodless, P.A. et al. (1996) Cell 85, 489-500.
3. Klemm, J.D. et al. (1998) Annu Rev Immunol 16, 569-92.
4. Kretzschmar, M. et al. (1997) Genes Dev 11, 984-95.
5. Whitman, M. (1998) Genes Dev 12, 2445-62.
6. Sapkota, G. et al. (2007) Mol Cell 25, 441-54.
7. Alarcón, C. et al. (2009) Cell 139, 757-69.

原厂资料：

Description

SignalSilence® Smad1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Smad1 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® Smad1 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

Bone morphogenetic proteins (BMPs) constitute a large family of signaling molecules that regulate a wide range of critical processes including morphogenesis, cell-fate determination, proliferation, differentiation, and apoptosis (1,2). BMP receptors are members of the TGF-β family of Ser/Thr kinase receptors. Ligand binding induces multimerization, autophosphorylation, and activation of these receptors (3-5). They subsequently phosphorylate Smad1 at Ser463 and Ser465 in the carboxy-terminal motif SSXS, as well as Smad5 and Smad8 at their corresponding sites. These phosphorylated Smads dimerize with the coactivating Smad4 and translocate to the nucleus, where they stimulate transcription of target genes (5).MAP kinases and CDKs 8 and 9 phosphorylate residues in the linker region of Smad1, including Ser206. The phosphorylation of Ser206 recruits Smurf1 to the linker region and leads to the degradation of Smad1 (6). Phosphorylation of this site also promotes Smad1 transcriptional action by recruiting YAP to the linker region (7).

1. Hogan, B.L. (1996) Genes Dev 10, 1580-94.
2. Hoodless, P.A. et al. (1996) Cell 85, 489-500.
3. Klemm, J.D. et al. (1998) Annu Rev Immunol 16, 569-92.
4. Kretzschmar, M. et al. (1997) Genes Dev 11, 984-95.
5. Whitman, M. (1998) Genes Dev 12, 2445-62.
6. Sapkota, G. et al. (2007) Mol Cell 25, 441-54.
7. Alarcón, C. et al. (2009) Cell 139, 757-69.

注意事项：

Storage: Smad1 siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC.