SignalSilence? YB1 siRNA II

• 产品编号：CST-6207S      品牌：CST       原厂货号：6207S
• 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^® YB1 siRNA 可以帮助研究者通过RNA干扰特异性地抑制YB1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^® YB1 siRNAII 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。Y-box结合蛋白1（YB1）属于一类进化上保守的，多功能Y-box蛋白，和单链DNA和RNA结合，调控转录，RNA代谢和蛋白质的合成（1）。YB1与多种基因启动子中的Y-box序列（TAACC）结合，并能正向或负向调节转录。YB1激活与增殖和癌症相关的基因，如周期蛋白A，B1，基质金属蛋白酶-2（MMP-2），及多药耐药性1（MDR1）基因（2-4）。YB1抑制与细胞死亡有关的基因，包括细胞的Fas相关死亡受体和p53抑癌基因（5-7）。它还与RNA剪切因子SRp30c互作并且能够稳定诱导T淋巴细胞的白细胞介素2的mRNA（8,9）。YB1蛋白主要定位于细胞质，少量存在于细胞核中；然而，核里面的YB1对促进细胞增殖十分关键。核转运受到细胞周期调控，在G1 / S期YB1蛋白质积累在细胞核中（2）。此外，细胞受到外部刺激，如高温和紫外线照射，或着用凝血酶，干扰素，胰岛素样生长因子（IGF-1）处理后可以诱导发生核转运（2,10）。IGF-1处理MCF7乳腺癌细胞株诱发Akt介导的YB1（Ser102）磷酸化，促进YB1的核转运及非贴壁依赖性生长（10）。YB1在许多恶性肿瘤组织中高表达，包括乳腺癌，非小细胞肺癌，卵巢癌，人骨肉瘤，大肠癌，恶性黑色素瘤。核YB1表达与过度增殖，耐药性，及肿瘤的不良预后相关（2,7,10）。

1. Matsumoto, K. and Wolffe, A.P. (1998) Trends Cell Biol. 8, 318-23.
2. Jurchott, K. et al. (2003) J. Biol. Chem. 278, 27988-96.
3. Mertens, P.R. et al. (1997) J. Biol. Chem. 272, 22905-12.
4. Uchiumi, T. et al. (1993) Cell Growth Differ. 4, 147-57.
5. Lasham, A. et al. (2000) Gene 252, 1-13.
6. Lasham, A. et al. (2003) J. Biol. Chem. 278, 35516-23.
7. Homer, C. et al. (2005) Oncogene 24, 8314-25.
8. Raffetseder, U. et al. (2003) J. Biol. Chem. 278, 18241-8.
9. Chen, C.Y. et al. (2000) Genes Dev. 14, 1236-48.
10. Sutherland, B.W. et al. (2005) Oncogene 24, 4281-92.

原厂资料：

Description

SignalSilence® YB1 siRNA from Cell Signaling Technology (CST) allows the researcher to specifically inhibit YB1 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 target protein expression in specified cell lines.

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 YB1 siRNA II 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 Y-box binding protein 1 (YB1) belongs to a family of evolutionarily conserved, multifunctional Y-box proteins that bind single-stranded DNA and RNA and function as regulators of transcription, RNA metabolism, and protein synthesis (1). YB1 binds to Y-box sequences (TAACC) found in multiple gene promoters and can positively or negatively regulate transcription. YB1 activates genes associated with proliferation and cancer, such as cyclin A, cyclin B1, matrix metalloproteinase-2 (MMP-2), and the multi-drug resistance 1 (MDR1) gene (2-4). YB1 represses genes associated with cell death, including the Fas cell death-associated receptor and the p53 tumor suppressor gene (5-7). It also interacts with the RNA-splicing factor SRp30c and stabilizes interleukin 2 mRNA upon induction of T lymphocytes by interleukin 2 (8,9). The majority of YB1 protein localizes to the cytoplasm, with a minor pool found in the nucleus; however, nuclear localization appears to be critical for its role in promoting proliferation. Nuclear translocation is cell cycle regulated, with YB1 protein accumulating in the nucleus during G1/S phase (2). In addition, nuclear translocation is induced in response to extracellular stimuli such as hyperthermia and UV irradiation, or treatment of cells with thrombin, interferons, or insulin-like growth factor (IGF-1) (2,10). Treatment of the MCF7 breast cancer cell line with IGF-1 results in Akt-mediated phosphorylation of YB1 on Ser102, which is required for nuclear translocation of YB1 and its ability to promote anchorage-independent growth (10). YB1 is overexpressed in many malignant tissues, including breast cancer, non-small cell lung carcinoma, ovarian adenocarcinomas, human osteosarcomas, colorectal carcinomas, and malignant melanomas. Nuclear YB1 expression correlates with high levels of proliferation, drug resistance, and poor tumor prognosis (2,7,10).

1. Matsumoto, K. and Wolffe, A.P. (1998) Trends Cell Biol. 8, 318-23.
2. Jurchott, K. et al. (2003) J. Biol. Chem. 278, 27988-96.
3. Mertens, P.R. et al. (1997) J. Biol. Chem. 272, 22905-12.
4. Uchiumi, T. et al. (1993) Cell Growth Differ. 4, 147-57.
5. Lasham, A. et al. (2000) Gene 252, 1-13.
6. Lasham, A. et al. (2003) J. Biol. Chem. 278, 35516-23.
7. Homer, C. et al. (2005) Oncogene 24, 8314-25.
8. Raffetseder, U. et al. (2003) J. Biol. Chem. 278, 18241-8.
9. Chen, C.Y. et al. (2000) Genes Dev. 14, 1236-48.
10. Sutherland, B.W. et al. (2005) Oncogene 24, 4281-92.

注意事项：

Storage: YB1 siRNA II is supplied in RNAse-free water. Aliquot
and store at -20ºC.