SignalSilence? SET8 siRNA I

• 产品编号：CST-13067S      品牌：CST       原厂货号：13067S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence^® SET8 siRNA I能够抑制人和猴子SET8 的表达。来自Cell Signaling Technology （CST）的SignalSilence^® SET8 siRNA I可以帮助研究者通过RNA干扰特异性抑制SET8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建议使用100 nM SET8 siRNA I进行转染，并在48-72小时后裂解细胞。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。每小瓶可供100次转染，每次转染量相当于在转染24孔板时，每孔总体积为300μl培养基中siRNA的终浓度为100nM。SET domain-containing lysine methyltransferase 8 (SET8)也称为PR/SET domain-containing protein 7 (PR/SET7)，它是一个组蛋白赖氨酸甲基转移酶家族成员，它们中每一个都包含一个保守催化的SET结构，该结构起初被鉴定在Drosophila Su[var]3-9、Enhancer of zeste和Trithorax蛋白中(1-3)。SET8是一个单亚单位激酶，它能使histone H4蛋白Lys20位点发生单甲基化，更适应在核小体结构底物上(1-3)。SET8蛋白水平和Histone H4蛋白Lys20位点甲基化是被细胞周期调节的，在S期处于增长期而到G2/M期达到最高峰(4,5)。SET8与PCNA蛋白相互作用，这与活化的DNA合成的位点有关联，并且对于在S期中DNA复制和基因组稳定性是需要的(5-7)。使用shRNA进行SET8的抑制可导致复制叉的阻止、双链DNA诱导的中断和在细胞周期的S和G2/M期中一个Chk1介导的细胞周期停滞 (6,7)。更进一步，SET8可使p53蛋白在Lys382位点甲基化，这可下调p53蛋白促凋亡和监测点激活功能(8)。在DNA损伤反应下，SET8表达水平下降，这允许p53去激活检测点和/或凋亡(8)。histone H4蛋白Lys20位点的甲基化和p53出现对于在S期中SET8的功能是非常重要的。

1. Fang, J. et al. (2002) Curr Biol 12, 1086-99.
2. Xiao, B. et al. (2005) Genes Dev 19, 1444-54.
3. Couture, J.F. et al. (2005) Genes Dev 19, 1455-65.
4. Rice, J.C. et al. (2002) Genes Dev 16, 2225-30.
5. Huen, M.S. et al. (2008) J Biol Chem 283, 11073-7.
6. Tardat, M. et al. (2007) J Cell Biol 179, 1413-26.
7. Jørgensen, S. et al. (2007) J Cell Biol 179, 1337-45.
8. Shi, X. et al. (2007) Mol Cell 27, 636-46.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence^® SET8 siRNA I will inhibit human and monkey SET8 expression.

Description

SignalSilence^® SET8 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit SET8 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 are rigorously tested in-house and have been shown to reduce 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 SET8 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.

Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

Background

SET domain-containing lysine methyltransferase 8 (SET8), also known as PR/SET domain-containing protein 7 (PR/SET7), is a member of a family of histone lysine methyltransferases, each of which contains a conserved catalytic SET domain originally identified in Drosophila Su[var]3-9, Enhancer of zeste, and Trithorax proteins (1-3). SET8 is a single-subunit enzyme that mono-methylates histone H4 on Lys20, preferably on nucleosomal substrates (1-3). SET8 protein levels and Histone H4 Lys20 methylation are cell cycle regulated, both increasing in S phase and peaking at G2/M phase (4,5). SET8 interacts with the PCNA protein, associates with sites of active DNA synthesis, and is required for DNA replication and genome stability during S phase (5-7). Inhibition of SET8 using shRNA or siRNA results in arrest of replication forks, induction of double-stranded DNA breaks, and a Chk1-mediated cell-cycle arrest in S and G2/M phases of the cell cycle (6,7). Furthermore, SET8 methylates p53 on Lys382, down regulating the pro-apoptotic and checkpoint activation functions of p53 (8). In response to DNA damage, SET8 expression levels decrease, allowing p53 to activate checkpoints and/or apoptosis (8). Both the methylation of histone H4 Lys20 and p53 appear to be important for the functions of SET8 in S phase.

1. Fang, J. et al. (2002) Curr Biol 12, 1086-99.
2. Xiao, B. et al. (2005) Genes Dev 19, 1444-54.
3. Couture, J.F. et al. (2005) Genes Dev 19, 1455-65.
4. Rice, J.C. et al. (2002) Genes Dev 16, 2225-30.
5. Huen, M.S. et al. (2008) J Biol Chem 283, 11073-7.
6. Tardat, M. et al. (2007) J Cell Biol 179, 1413-26.
7. Jørgensen, S. et al. (2007) J Cell Biol 179, 1337-45.
8. Shi, X. et al. (2007) Mol Cell 27, 636-46.

注意事项：

Storage: SET8 siRNA I is supplied in RNAse-free water. Aliquot
and store at -20ºC.