SignalSilence? Aurora A/AIK siRNA I

• 产品编号：CST-8883S      品牌：CST       原厂货号：8883S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

CST的SignalSilence® Aurora A/AIK siRNA I 允许研究人员采用RNA干扰（一种通过传递外源性的双链RNA分子到细胞中从而能选择性的沉默基因表达的方法）技术特异性的抑制Aurora A/AIK蛋白表达。CST所有的SignalSilence® siRNA产品都经过了严格的内部测试并采用Western分析证明其可以有效降低靶蛋白的表达。为了保证适当的结合效率，我们采用三苯甲基分析的方法逐个碱基的监测寡核苷酸的合成。随后通过亲和固相提取纯化寡核苷酸。退火的双链RNA进一步通过质谱分析来验证双链复合体的确切成分。每一批产品和前一批都要通过质谱分析进行比较以确定批次间最大限度的一致性。CST建议使用100 nM SignalSilence® Aurora A/AIK siRNA I转染48-72小时后裂解细胞。转染的具体步骤请参考转染试剂制造商提供的操作标准。使用过程中有任何问题请随时联系CST。 极光激酶属于有丝分裂过程中高度保守的丝氨酸/苏氨酸激酶家族，在哺乳动物中已确定了三个成员：极光激酶A，极光激酶B和极光激酶C(1,2)。时空表达模式和有丝分裂细胞中极光激酶亚细胞定位的研究表明其和有丝分裂结构的关系。极光激酶功能的影响跨越G2期和细胞分裂期并可能涉及到关键的细胞周期事件，如中心体复制、染色体双向定位和隔离、卵裂沟定位和内移(3)。极光激酶A可以在中心体(沿纺锤体微管方向)和有丝分裂增殖细胞的胞质中被检测到。极光激酶A蛋白水平在G1和S期较低而在细胞周期的G2/M期达峰值。极光激酶A催化结构域苏氨酸(288位)的磷酸化能够增强激酶的活性。极光激酶A参与中心体分离、成熟和纺锤体组装及稳定。极光激酶B的蛋白表达同样在细胞周期的G2/M期达峰值; 极光激酶B的激酶活性在分裂中期到有丝分裂结束的过渡阶段最高。在重新定位到有丝分裂后期纺锤体之前，极光激酶B和分裂前期的染色体密切相关。极光激酶B通过控制微管-着丝粒附着和细胞质分裂调节染色体分离。G2/M期过渡时极光激酶A和极光激酶B的表达紧密配合组蛋白H3的磷酸化(4,5); 多种人类癌症中都发现了这些激酶的过度表达(2,4)。极光激酶C集中于有丝分裂后期到细胞质分裂期的中心体中，而且其mRNA和蛋白质水平在G2/M期达峰值。虽然典型的极光激酶C的表达仅限于睾丸，但是在各种肿瘤细胞株中都可以检测到其过度表达(6)。

1. Warner, S.L. et al. (2003) Mol Cancer Ther 2, 589-95.
2. Katayama, H. et al. (2003) Cancer Metastasis Rev 22, 451-64.
3. Andrews, P.D. et al. (2003) Curr Opin Cell Biol 15, 672-83.
4. Pascreau, G. et al. (2003) Prog Cell Cycle Res 5, 369-74.
5. Crosio, C. et al. (2002) Mol Cell Biol 22, 874-85.
6. Kimura, M. et al. (1999) J Biol Chem 274, 7334-40.

原厂资料：

Description

SignalSilence® Aurora A/AIK siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Aurora A/AIK 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® Aurora A/AIK 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

Aurora kinases belong to a highly conserved family of mitotic serine/threonine kinases with three members identified among mammals: Aurora A, B, and C (1,2). Studies on the temporal expression pattern and subcellular localization of Aurora kinases in mitotic cells suggest an association with mitotic structure. Their functional influences span from G2 phase to cytokinesis and may be involved in key cell cycle events such as centrosome duplication, chromosome bi-orientation and segregation, cleavage furrow positioning, and ingression (3). Aurora A is detected at the centrosomes, along mitotic spindle microtubules, and in the cytoplasm of mitotically proliferating cells. Aurora A protein levels are low during G1 and S phases and peak during the G2/M phase of the cell cycle. Phosphorylation of Aurora A at Thr288 in its catalytic domain increases kinase activity. Aurora A is involved in centrosome separation, maturation, and spindle assembly and stability. Expression of Aurora B protein also peaks during the G2/M phase of the cell cycle; Aurora B kinase activity peaks at the transition from metaphase to the end of mitosis. Aurora B associates with chromosomes during prophase prior to relocalizing to the spindle at anaphase. Aurora B regulates chromosome segregation through the control of microtubule-kinetochore attachment and cytokinesis. Expression of both Aurora A and Aurora B during the G2/M phase transition is tightly coordinated with histone H3 phosphorylation (4,5); overexpression of these kinases is seen in a variety of human cancers (2,4). Aurora C localizes to the centrosome from anaphase to cytokinesis and both mRNA and protein levels peak during G2/M phase. Although typical Aurora C expression is limited to the testis, overexpression of Aurora C is detected in various cancer cell lines (6). 

1. Warner, S.L. et al. (2003) Mol Cancer Ther 2, 589-95.
2. Katayama, H. et al. (2003) Cancer Metastasis Rev 22, 451-64.
3. Andrews, P.D. et al. (2003) Curr Opin Cell Biol 15, 672-83.
4. Pascreau, G. et al. (2003) Prog Cell Cycle Res 5, 369-74.
5. Crosio, C. et al. (2002) Mol Cell Biol 22, 874-85.
6. Kimura, M. et al. (1999) J Biol Chem 274, 7334-40.

注意事项：

Storage: Aurora A/AIK siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC.