SignalSilence? SirT6 siRNA I

• 产品编号：CST-9390S      品牌：CST       原厂货号：9390S
• 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® SirT6 siRNA I 可以通过RNA干扰特异性抑制SirT6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 SignalSilence® SirT6 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。沉默信息调节因子（SIR2）基因家族是一类高度保守的基因，能够编码烟酰胺腺嘌呤二核苷酸（NAD）依赖性的去乙酰化酶，亦称III类组蛋白去乙酰化酶。在这些基因中，酿酒酵母Sir2是最早被发现，并且了解得最清楚的。Sir2 参与了交配型基因座的沉默，端粒维持，DNA损伤应答和细胞衰老(1)。SirT6，是Sir2在哺乳动物中的同源基因，是一种核仁、染色质相关蛋白，能够通过碱基修复（BER）信号通路以正常维持基因组的完整性（2-4）。BER通路可以修复内源烷化、氧化和脱氨基作用中自发的单链DNA损伤。SirT6缺乏的小鼠，对DNA损伤药剂的敏感性明显增强，包括烷化药剂MMS和H2O2 (2)。另外，这些小鼠中还显示出基因组的不稳定性，其断裂染色体、分离的着丝粒以及缺口的出现频率明显增加(2)。SirT6可能通过对通路中DNA Polβ或其他核心组分的去乙酰化作用来调控BER通路（2）。

1. Guarente, L. (1999) Nat Genet 23, 281-5.
2. Mostoslavsky, R. et al. (2006) Cell 124, 315-29.
3. Liszt, G. et al. (2005) J Biol Chem 280, 21313-20.
4. Michishita, E. et al. (2005) Mol Biol Cell 16, 4623-35.

原厂资料：

Description

SignalSilence® SirT6 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit SirT6 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® SirT6 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

The Silent Information Regulator (Sir2) family of genes is a highly conserved group of genes that encode nicotinamide adenine dinucleotide (NAD)-dependent protein deacetylases, also known as class III histone deacetylases. The first discovered and best characterized of this family is Saccharomyces cerevisiae Sir2, which is involved in silencing of mating type loci, telomere maintenance, DNA damage response, and cell aging (1). SirT6, a mammalian homolog of Sir2, is a nuclear, chromatin-associated protein that promotes the normal maintenance of genome integrity mediated by the base excision repair (BER) pathway (2-4). The BER pathway repairs single-stranded DNA lesions that arise spontaneously from endogenous alkylation, oxidation, and deamination events. SirT6 deficient mice show increased sensitivity to DNA-damaging agents, including the alkylating agents MMS and H2O2 (2). In addition, these mice show genome instability with increased frequency of fragmented chromosomes, detached centromeres, and gaps (2). SirT6 may regulate the BER pathway by deacetylating DNA Polβ or other core components of the pathway (2).

1. Guarente, L. (1999) Nat Genet 23, 281-5.
2. Mostoslavsky, R. et al. (2006) Cell 124, 315-29.
3. Liszt, G. et al. (2005) J Biol Chem 280, 21313-20.
4. Michishita, E. et al. (2005) Mol Biol Cell 16, 4623-35.

注意事项：

Storage: SirT6 siRNA I is supplied in RNAse-free water. Aliquot
and store at -20ºC.