SignalSilence? BRCA1 siRNA II

• 产品编号：CST-12642S      品牌：CST       原厂货号：12642S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence® BRCA1 siRNA II可以抑制人和猴的BRCA1基因表达。Cell Signaling Technology (CST) 公司的SignalSilence® BRCA1 siRNA II帮助研究人员使用RNA干扰技术特异性抑制BRCA1表达，该技术通过向细胞内导入双链RNA分子以选择性的抑制某些基因的表达。所有CST公司的SignalSilence® siRNA都经过了严格的内部测试，western分析显示能够能够降低目标蛋白的表达。寡核苷酸合成经过了三苯甲基的逐碱基分析以确保耦合效率。随后经过亲和固相萃取纯化。退火的RNA双链在经过质谱分析以确保双链的准确性。新批次都会经过质谱分析与之前的批次进行比对以保证不同批次之间的稳定性。CST建议转染100 nM SignalSilence® BRCA1 siRNA II，48-72小时后裂解细胞。转染的步骤请参阅转染试剂的说明书。关于使用问题，可以随时咨询CST。 每管试剂能进行100次转染，该转染次数以总体积为300μl 的1个24孔板孔中siRNA终浓度100 nM来进行计算。乳腺癌易感蛋白BRCA1和BRCA2在遗传性乳腺癌和卵巢癌病人体内经常会发生突变，在DNA损伤，修复，细胞周期进程，转录，泛素化和细胞凋亡等多个过程中都发挥了作用（1-4）。已经发现BRCA2对于Rad51定位到双链损伤（DSBs）区域是必须的，细胞缺乏BRCA1和BRCA2不能通过Rad51依赖的同源重组（HR）修复DSBs（5）。已经在BRCA1蛋白上发现了多个DNA损伤诱导的磷酸化位点，包括Ser988, 1189, 1387, 1423, 1457, 1524 和1542，并且激酶以一种细胞周期依赖的方式被激活，包括Aurora A 和 CDK2,都可以分别磷酸化BRCA1的308和1497位丝氨酸（6-10）。BRCA1 的3291位丝氨酸发生的细胞周期依赖的磷酸化是通过CDKs进行的，已经被认为是一种阻止羧基末端Rad51结合区域以促进细胞通过S期中止HR的机制（11）。

1. Rahman, N. and Stratton, M.R. (1998) Annu Rev Genet 32, 95-121.
2. Gayther, S.A. et al. (1999) Am J Hum Genet 65, 1021-9.
3. Kerr, P. and Ashworth, A. (2001) Curr Biol 11, R668-76.
4. Scully, R. and Livingston, D.M. (2000) Nature 408, 429-32.
5. Tutt, A. and Ashworth, A. (2002) Trends Mol Med 8, 571-6.
6. Okada, S. and Ouchi, T. (2003) J Biol Chem 278, 2015-20.
7. Cortez, D. et al. (1999) Science 286, 1162-1166.
8. Xu, B. et al. (2002) Cancer Res 62, 4588-91.
9. Ouchi, M. et al. (2004) J Biol Chem 279, 19643-8.
10. Ruffner, H. et al. (1999) Mol Cell Biol 19, 4843-54.
11. Esashi, F. et al. (2005) Nature 434, 598-604.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence® BRCA1 siRNA II inhibits human and monkey BRCA1 expression.

Description

SignalSilence® BRCA1 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit BRCA1 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® BRCA1 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. 
 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 breast cancer susceptibility proteins BRCA1 and BRCA2 are frequently mutated in cases of hereditary breast and ovarian cancers and have roles in multiple processes related to DNA damage, repair, cell cycle progression, transcription, ubiquitination, and apoptosis (1-4). BRCA2 has been shown to be required for localization of Rad51 to sites of double stranded breaks (DSBs) in DNA, and cells lacking BRCA1 and BRCA2 cannot repair DSBs through the Rad51-dependent process of homologous recombination (HR) (5). Numerous DNA damage-induced phosphorylation sites on BRCA1 have been identified, including Ser988, 1189, 1387, 1423, 1457, 1524 and 1542, and kinases activated in a cell cycle-dependent manner, including Aurora A and CDK2, can also phosphorylate BRCA1 at Ser308 and Ser1497, respectively (6-10). Cell cycle-dependent phosphorylation of BRCA2 at Ser3291 by CDKs has been proposed as a mechanism to switch off HR as cells progress beyond S-phase by blocking the carboxy terminal Rad51 binding site (11).

1. Rahman, N. and Stratton, M.R. (1998) Annu Rev Genet 32, 95-121.
2. Gayther, S.A. et al. (1999) Am J Hum Genet 65, 1021-9.
3. Kerr, P. and Ashworth, A. (2001) Curr Biol 11, R668-76.
4. Scully, R. and Livingston, D.M. (2000) Nature 408, 429-32.
5. Tutt, A. and Ashworth, A. (2002) Trends Mol Med 8, 571-6.
6. Okada, S. and Ouchi, T. (2003) J Biol Chem 278, 2015-20.
7. Cortez, D. et al. (1999) Science 286, 1162-1166.
8. Xu, B. et al. (2002) Cancer Res 62, 4588-91.
9. Ouchi, M. et al. (2004) J Biol Chem 279, 19643-8.
10. Ruffner, H. et al. (1999) Mol Cell Biol 19, 4843-54.
11. Esashi, F. et al. (2005) Nature 434, 598-604.

注意事项：

Storage: BRCA1 siRNA II is supplied in RNAse-free water.
Aliquot and store at -20ºC.