SignalSilence? Bmi1 siRNA I

• 产品编号：CST-6442S      品牌：CST       原厂货号：6442S
• 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)公司的SignalSilence® VASP siRNA I可以帮助研究者通过RNA干扰去特异性抑制Bmi1的蛋白表达，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公司推荐使用100 nM Bmi1 siRNAI 进行转染，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。polycomb group (PcG)蛋白有助于细胞身份的维持、干细胞自我更新、细胞周期调节和肿瘤形成，这些功能通过维持一些基因的沉默，而这些基因是促进细胞系特异性、细胞死亡和细胞周期停滞(1-4)。PcG蛋白存在两个复合物，它们配合通过表观遗传染色质修饰去维持长期的基因沉默。通过DNA结合转录因子和组蛋白H3在Lys27位点的甲基化，第一个复合物EED-EZH2被招募到基因上。这个组蛋白甲基转移酶活性需要该复合物的Ezh2、Eed和Suz12亚基(5)。组蛋白H3在Lys27位点的甲基化促进第二个复合物PRC1的招募，其能使组蛋白H2A在Lys119位点泛素化(6)。Bmi1蛋白是PRC1复合物的一个组成成分，该蛋白可与Ring1一起强有力的提高了Ring2催化亚基的E3泛素连接酶活性(7)。Bmi1蛋白在调节细胞增殖和衰老中起到重要作用，这主要通过p16 INK4A和p19 ARF基因的抑制，并且Bmi1蛋白对于成人造血和神经干细胞的维持是必须的(3,4,8-10)。

1. Boyer, L.A. et al. (2006) Nature 441, 349-53.
2. Lee, T.I. et al. (2006) Cell 125, 301-13.
3. Park, I.K. et al. (2003) Nature 423, 302-5.
4. Molofsky, A.V. et al. (2003) Nature 425, 962-7.
5. Cao, R. and Zhang, Y. (2004) Mol Cell 15, 57-67.
6. Wang, H. et al. (2004) Nature 431, 873-8.
7. Cao, R. et al. (2005) Mol Cell 20, 845-54.
8. Molofsky, A.V. et al. (2005) Genes Dev 19, 1432-7.
9. Jacobs, J.J. et al. (1999) Nature 397, 164-8.
10. Jacobs, J.J. et al. (1999) Genes Dev 13, 2678-90.

原厂资料：

Description

SignalSilence® Bmi1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Bmi1 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 Bmi1 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 polycomb group (PcG) of proteins contributes to the maintenance of cell identity, stem cell self-renewal, cell cycle regulation, and oncogenesis by maintaining the silenced state of genes that promote cell lineage specification, cell death, and cell-cycle arrest (1-4). PcG proteins exist in two complexes that cooperate to maintain long-term gene silencing through epigenetic chromatin modifications. The first complex, EED-EZH2, is recruited to genes by DNA-binding transcription factors and methylates histone H3 on Lys27. This histone methyl-transferase activity requires the Ezh2, Eed, and Suz12 subunits of the complex (5). Histone H3 methylation at Lys27 facilitates the recruitment of the second complex, PRC1, which ubiquitinylates histone H2A on Lys119 (6). Bmi1 is a component of the PRC1 complex, which together with Ring1 strongly enhances the E3 ubiquitin ligase activity of the Ring2 catalytic subunit (7). Bmi1 plays an important role in the regulation of cell proliferation and senescence through repression of the p16 INK4A and p19 ARF genes and is required for maintenance of adult hematopoietic and neural stem cells (3,4,8-10).

1. Boyer, L.A. et al. (2006) Nature 441, 349-53.
2. Lee, T.I. et al. (2006) Cell 125, 301-13.
3. Park, I.K. et al. (2003) Nature 423, 302-5.
4. Molofsky, A.V. et al. (2003) Nature 425, 962-7.
5. Cao, R. and Zhang, Y. (2004) Mol Cell 15, 57-67.
6. Wang, H. et al. (2004) Nature 431, 873-8.
7. Cao, R. et al. (2005) Mol Cell 20, 845-54.
8. Molofsky, A.V. et al. (2005) Genes Dev 19, 1432-7.
9. Jacobs, J.J. et al. (1999) Nature 397, 164-8.
10. Jacobs, J.J. et al. (1999) Genes Dev 13, 2678-90.

注意事项：

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