SignalSilence? FoxO3a siRNA I

• 产品编号：CST-6302S      品牌：CST       原厂货号：6302S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

CST公司SignalSilence ® FoxO3a siRNA I使得研究者可以通过使用RNA干扰来特异性抑制FoxO3a的表达。RNA干扰是一种通过向细胞中输入双链RNA分子来选择性的抑制基因的表达方法。CST公司的所有SignalSilence ® siRNA 产品已经过严格的内部测试，western blot检测显示其可以降低靶蛋白的表达。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建议用100 nM FoxO3a siRNA I转染48-72 小时后裂解细胞。对于转染步骤，参见转染试剂生产商提供的说明书。如果有问题请随时与CST联系。Forkhead转录因子家族参与横纹肌肉瘤和急性白血病的发生（1-3）。在这个家族中，三个成员（FoxO1，FoxO4和FoxO3a）与线虫的同源基因DAF-16具有序列相似度，而DAF-16介导包括IGFR1，PI3K和Akt在内的信号通路（4-6）。活化的forkhead成员作为肿瘤抑制因子，促进细胞周期阻滞和凋亡。FOXO家族任何成员的表达上调都会激活细胞周期抑制剂p27 Kip1。Forkhead转录因子也参与TGF-β介导的p21 CIP1上调，而这个上调过程可以被PI3K负调控（7）。forkhead转录因子被Akt在Thr24，Ser256以及Ser319磷酸化后而失活，导致出核转运与转录因子活性的抑制（8），最终导致增殖上调。Forkhead转录因子活性也可以被去乙酰化酶sirtuin(SirT1)抑制（9）。

1. Anderson, M.J. et al. (1998) Genomics 47, 187-99.
2. Galili, N. et al. (1993) Nat Genet 5, 230-5.
3. Borkhardt, A. et al. (1997) Oncogene 14, 195-202.
4. Nakae, J. et al. (1999) J Biol Chem 274, 15982-5.
5. Rena, G. et al. (1999) J Biol Chem 274, 17179-83.
6. Guo, S. et al. (1999) J Biol Chem 274, 17184-92.
7. Seoane, J. et al. (2004) Cell 117, 211-23.
8. Arden, K.C. (2004) Mol Cell 14, 416-8.
9. Yang, Y. et al. (2005) EMBO J 24, 1021-32.

原厂资料：

Description

SignalSilence® FoxO3a siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit FoxO3a 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 FoxO3a 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

The Forkhead family of transcription factors is involved in tumorigenesis of rhabdomyosarcoma and acute leukemias (1-3). Within the family, three members (FoxO1, FoxO4, and FoxO3a) have sequence similarity to the nematode orthologue DAF-16, which mediates signaling via a pathway involving IGFR1, PI3K, and Akt (4-6). Active forkhead members act as tumor suppressors by promoting cell cycle arrest and apoptosis. Increased expression of any FoxO member results in the activation of the cell cycle inhibitor p27 Kip1. Forkhead transcription factors also play a part in TGF-β-mediated upregulation of p21 Cip1, a process negatively regulated through PI3K (7). Increased proliferation results when forkhead transcription factors are inactivated through phosphorylation by Akt at Thr24, Ser256, and Ser319, which results in nuclear export and inhibition of transcription factor activity (8). Forkhead transcription factors can also be inhibited by the deacetylase sirtuin (SirT1) (9).

1. Anderson, M.J. et al. (1998) Genomics 47, 187-99.
2. Galili, N. et al. (1993) Nat Genet 5, 230-5.
3. Borkhardt, A. et al. (1997) Oncogene 14, 195-202.
4. Nakae, J. et al. (1999) J Biol Chem 274, 15982-5.
5. Rena, G. et al. (1999) J Biol Chem 274, 17179-83.
6. Guo, S. et al. (1999) J Biol Chem 274, 17184-92.
7. Seoane, J. et al. (2004) Cell 117, 211-23.
8. Arden, K.C. (2004) Mol Cell 14, 416-8.
9. Yang, Y. et al. (2005) EMBO J 24, 1021-32.

注意事项：

Storage: FoxO3a siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC.