SignalSilence? p16 INK4A siRNA I

• 产品编号：CST-6598S      品牌：CST       原厂货号：6598S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

CST的SignalSilence® p16 INK4A siRNA I使得研究人员可以使用RNA干扰的方法特异性抑制p16 INK4A的表达，RNA干扰是一种通过将双链RNA分子转入细胞从而选择性沉默基因表达的方法。CST的所有SignalSilence® siRNA产品都通过了严格的内部测试，并使用western blot方法检测其减少目标蛋白表达的效用。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。细胞周期依赖性蛋白激酶(CDKs)的激活部分经由周期蛋白复合体的形成。例如，CDK4和CDK6联合D-型周期蛋白并磷酸化Rb蛋白。这种磷酸化使细胞进入S期的必需事件(1)。CDK4 (INK4)家族抑制剂包括p15 INK4B，p16 INK4A，p18 INK4C 和p19 INK4D。p18被认为在体能够发挥单纯不充分肿瘤抑制基因的功能(2)。所有的INK4蛋白是由32个氨基酸锚蛋白模体组成并可以选择性抑制CDK4/6的活性。P18的突变分析表明第三个锚蛋白和第四个锚蛋白重复序列的氨基末端部分介导了CDK4/6的结合(3)。INK4家族成员的相互作用可能是通过与CDK4/6形成的二元复合物或者是与周期蛋白-D结合的CDK4/6形成的三元复合物，最终导致细胞周期进程的抑制(4,5)。 p16 INK4A能够直接抑制周期蛋白-D的活性从而抑制S-期进入(6,7)。同样的，p16 INK4A的表达通常和细胞衰老有关，而在人类恶性肿瘤中可以频繁观察到p16 INK4A基因的破坏。

1. Lukas, J. et al. (1996) Mol. Cell. Biol. 16, 6917-6925.
2. Bai, F. et al. (2003) Mol. Cell. Biol. 23, 1269-1277.
3. Noh, S.J. et al. (1999) Cancer Res. 59, 558-564.
4. Guan, K.L. et al. (1994) Genes Dev. 8, 2939-2952.
5. Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681.
6. Sherr, C.J. (2001) Nat Rev Mol Cell Biol 2, 731-7.
7. Lowe, S.W. and Sherr, C.J. (2003) Curr Opin Genet Dev 13, 77-83.

原厂资料：

Description

SignalSilence® p16 INK4A siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit p16 INK4A 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 p16 INK4A 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

Cyclin-dependent kinases (CDKs) are activated in part by forming complexes with cyclins. For example, CDK4 and CDK6 associate with the D-type cyclins and phosphorylate the retinoblastoma protein. This phosphorylation is a necessary event for cells to enter S-phase (1). The inhibitors of CDK4 (INK4) family include p15 INK4B, p16 INK4A, p18 INK4C and p19 INK4D. p18 has been shown to function as a haploinsufficient tumor suppressor in vivo (2). All INK4 proteins are composed of 32 amino acid ankyrin motifs and selectively inhibit CDK4/6 activity. Mutational analyses of p18 implicate the third and the amino-terminal portion of the fourth ankyrin repeat in mediating binding to CDK4/6 (3). The interaction of INK4 family members can be a binary complex with CDK4/6 or ternary complex with cyclin D-bound CDK4/6 and ultimately results in the inhibition of cell cycle progression (4,5).p16 INK4A directly inhibits the activity of cyclin D, thereby inhibiting S-phase entry (6,7). As such, expression of p16 INK4A is commonly associated with cellular senescence, and disruption of the p16 INK4A gene is frequently observed in human cancers. 

1. Lukas, J. et al. (1996) Mol. Cell. Biol. 16, 6917-6925.
2. Bai, F. et al. (2003) Mol. Cell. Biol. 23, 1269-1277.
3. Noh, S.J. et al. (1999) Cancer Res. 59, 558-564.
4. Guan, K.L. et al. (1994) Genes Dev. 8, 2939-2952.
5. Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681.
6. Sherr, C.J. (2001) Nat Rev Mol Cell Biol 2, 731-7.
7. Lowe, S.W. and Sherr, C.J. (2003) Curr Opin Genet Dev 13, 77-83.

注意事项：

Storage: p16 INK4A siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC