SignalSilence? JMJD2B siRNA I

• 产品编号：CST-7435S      品牌：CST       原厂货号：7435S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

Cell Signaling Technology (CST)公司的SignalSilence® JMJD2B siRNA I可以帮助研究者通过RNA干扰特异性地抑制JMJD2B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 SignalSilence® JMJD2B siRNA I 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。组蛋白赖氨酸的甲基化水平对于基因组的活化和非活化区域的形成是主要决定因素，并且在发育期间对于基因组的正确进程起着关键作用(1,2)。包含蛋白质的Jumonji C (JmjC)区域代表最大的潜在组蛋白去乙酰化酶蛋白(3)。JmjC区域通过氧化反应能催化单、双和三甲基化的赖氨酸残基的去乙甲基化，这种氧化反应需要铁离子和α-酮戊二酸(3)。基于同源性，人源和小鼠都包含至少30种这样蛋白质，这能够被分为7个不同的家族(3)。JMJD2 (Jumonji domain-containing protein 2)家族也称为JHDM3 (JmjC domain-containing histone demethylation protein 3)家族，包含四个成员：JMJD2A/JHDM3A、JMJD2B/JHDM3B、JMJD2C/JHDM3C和JMJD2D/JHDM3D。除了JmJC结构域之外，这些蛋白质也包含JmJN、PHD和Tudor结构域，而后者基因证明能结合到甲基化的histone H3蛋白Lys4和Lys9 位点，以及甲基化的histone H4蛋白Lys20位点上(4,5)。研究证明JMJD2蛋白可以使双核三甲基化的histone H3蛋白Lys9和Lys36位点上去甲基化，并且具有转录的激活和抑制因子的功能(6-11)。JMJD2A、JMJD2C 和JMJD2D作为在前列腺肿瘤细胞中雄激素受体的共激活因子(7)。与此相反，JMJD2A也与Rb和N-CoR共抑制因子复合物有关联，并且对于靶基因的转录抑制是必须的(8,9)。在着丝点周围异染色质上JMJD2B蛋白抑制histone H3蛋白Lys9位点三甲基化(10)。JMJD2C也称为GASC1，它在鳞状细胞癌和转移性肺癌中扩增，并且JMJD2C蛋白表达的抑制减少了细胞扩增(11,12)。JMJD2C蛋白也被鉴定作为Oct-4的下游靶蛋白，并且在胚胎干细胞中对于自我更新的调节起到重要作用(13)。

1. Kubicek, S. et al. (2006) Ernst Schering Res Found Workshop , 1-27.
2. Lin, W. and Dent, S.Y. (2006) Curr Opin Genet Dev 16, 137-42.
3. Klose, R.J. et al. (2006) Nat Rev Genet 7, 715-27.
4. Chen, Z. et al. (2007) Proc Natl Acad Sci USA 104, 10818-23.
5. Lee, J. et al. (2008) Nat Struct Mol Biol 15, 109-11.
6. Whetstine, J.R. et al. (2006) Cell 125, 467-81.
7. Shin, S. and Janknecht, R. (2007) Biochem Biophys Res Commun 359, 742-6.
8. Gray, S.G. et al. (2005) J Biol Chem 280, 28507-18.
9. Zhang, D. et al. (2005) Mol Cell Biol 25, 6404-14.
10. Fodor, B.D. et al. (2006) Genes Dev 20, 1557-62.
11. Cloos, P.A. et al. (2006) Nature 442, 307-11.
12. Italiano, A. et al. (2006) Cancer Genet Cytogenet 167, 122-30.
13. Loh, Y.H. et al. (2007) Genes Dev 21, 2545-57.

原厂资料：

Description

SignalSilence® JMJD2B siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit JMJD2B 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® JMJD2B 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 methylation state of lysine residues in histone proteins is a major determinant of the formation of active and inactive regions of the genome and is crucial for proper programming of the genome during development (1,2). Jumonji C (JmjC) domain-containing proteins represent the largest class of potential histone demethylase proteins (3). The JmjC domain can catalyze the demethylation of mono-, di-, and tri-methyl lysine residues via an oxidative reaction that requires iron and α-ketoglutarate (3). Based on homology, both humans and mice contain at least 30 such proteins, which can be divided into 7 separate families (3). The jumonji domain-containing protein 2 (JMJD2) family, also known as the JmjC domain-containing histone demethylation protein 3 (JHDM3) family, contains four members: JMJD2A/JHDM3A, JMJD2B/JHDM3B, JMJD2C/JHDM3C, and JMJD2D/JHDM3D. In addition to the JmjC domain, these proteins also contain JmjN, PHD, and tudor domains, the latter of which has been shown to bind to methylated histone H3 at Lys4 and Lys9, and methylated histone H4 at Lys20 (4,5). JMJD2 proteins have been shown to demethylate di- and tri-methyl histone H3 at Lys9 and Lys36 and function as both activators and repressors of transcription (6-11). JMJD2A, JMJD2C, and JMJD2D function as coactivators of the androgen receptor in prostate tumor cells (7). In contrast, JMJD2A also associates with Rb and NCoR corepressor complexes and is necessary for transcriptional repression of target genes (8,9). JMJD2B antagonizes histone H3 Lys9 tri-methylation at pericentric heterochromatin (10). JMJD2C, also known as GASC1, is amplified in squamous cell carcinomas and metastatic lung carcinoma and inhibition of JMJD2C expression decreases cell proliferation (11,12). JMJD2C has also been identified as a downstream target of Oct-4 and is critical for the regulation of self-renewal in embryonic stem cells (13).

1. Kubicek, S. et al. (2006) Ernst Schering Res Found Workshop , 1-27.
2. Lin, W. and Dent, S.Y. (2006) Curr Opin Genet Dev 16, 137-42.
3. Klose, R.J. et al. (2006) Nat Rev Genet 7, 715-27.
4. Chen, Z. et al. (2007) Proc Natl Acad Sci USA 104, 10818-23.
5. Lee, J. et al. (2008) Nat Struct Mol Biol 15, 109-11.
6. Whetstine, J.R. et al. (2006) Cell 125, 467-81.
7. Shin, S. and Janknecht, R. (2007) Biochem Biophys Res Commun 359, 742-6.
8. Gray, S.G. et al. (2005) J Biol Chem 280, 28507-18.
9. Zhang, D. et al. (2005) Mol Cell Biol 25, 6404-14.
10. Fodor, B.D. et al. (2006) Genes Dev 20, 1557-62.
11. Cloos, P.A. et al. (2006) Nature 442, 307-11.
12. Italiano, A. et al. (2006) Cancer Genet Cytogenet 167, 122-30.
13. Loh, Y.H. et al. (2007) Genes Dev 21, 2545-57.

注意事项：

Storage: SignalSilence ® JMJD2B siRNA I is supplied in RNAse-
free water. Aliquot and store at -20°C.