SignalSilence? CDK5 siRNA I

• 产品编号：CST-6216S      品牌：CST       原厂货号：6216S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence® CDK5 siRNA I 会抑制人、小鼠和大鼠的CDK5表达。Cell Signaling Technology (CST)公司生产的SignalSilence® CDK5 siRNA I使得研究者可以使用RNA干扰技术特异性的抑制CDK5的表达。这种技术通过将双链RNA分子送入细胞而选择性的沉默某个基因的表达。SignalSilence® CST的siRNA产品已经在内部经过严格测试，经western分析确能减少目标蛋白的表达。通过三苯分析对寡核苷酸合成进行逐个碱基的监测，确保了正确的耦联效率。然后用亲和固相萃取纯化寡核苷酸。退火的RNA进一步通过质谱分析，以验证其确切成分。每批都将和以前批次进行质谱比对，以确保批次间最大的一致性。CST公司推荐您在细胞裂解前用100nM CDK5 siRNA I进行48-72小时的转染。转染的流程请遵循转染试剂制造商提供的操作流程。使用中如有任何问题，请与CST联系。细胞周期蛋白依赖性激酶（CDKs）是丝氨酸/苏氨酸激酶，它们被周期蛋白cyclins激活，控制真核细胞周期的进程。CDK5与其家庭成员分享同源性很高的序列，它被认为主要是在有丝分裂后的神经元中调节细胞结构。与周期蛋白cyclins类似，调节亚基p35和p39与CDK5结合并激活CDK5，尽管它们缺乏序列同源性。CDK5是广泛表达的，由于有丝分裂后神经元具有窄谱的表达P35和P39，其主要在中枢神经系统中具有高水平的激酶活性。大量CDK5的底物已被确定，虽然尚无某个底物被特异性的归于P35或P39。CDK5的底物包括p35, PAK1, Src, β-catenin, tau, neurofilament-H, neurofilament-M, synapsin1, APP, DARPP32, PP1-inhibitor 和 Rb。p35被泛素 - 蛋白酶体途径迅速降解（T1/ 2 <20分钟）（1）。然而，P35的稳定性增加时CDK5激酶活性降低，可能是因为CDK5在P35的Thr138位点磷酸化水平减少所造成（2）。毒性损害后calpain造成p35蛋白水解产生p25，导致CDK5长期被p25激活。P25积聚在神经退行性疾病如阿尔茨海默病和肌萎缩性侧索硬化症（ALS）中被发现（3,4）。

1. Dhavan, R. and Tsai, L.H. (2001) Nat Rev Mol Cell Biol 2, 749-59.
2. Patrick, G.N. et al. (1998) J Biol Chem 273, 24057-64.
3. Lee, M.S. et al. (2000) Nature 405, 360-4.
4. Kusakawa, G. et al. (2000) J Biol Chem 275, 17166-72.

原厂资料：

Homology

Species predicted to react based on 100% sequence homology: Mouse, Rat

Specificity / Sensitivity

SignalSilence® CDK5 siRNA I will inhibit human, mouse and rat CDK5 expression.

Description

SignalSilence® CDK5 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit CDK5 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 CDK5 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 serine/threonine kinases that are activated by cyclins and govern eukaryotic cell cycle progression. While CDK5 shares high sequence homology with its family members, it is thought mainly to function in postmitotic neurons to regulate the cytoarchitecture of these cells. Analogous to cyclins, the regulatory subunits p35 and p39 associate with and activate CDK5 despite the lack of sequence homology. CDK5 is ubiquitously expressed, with high levels of kinase activity detected primarily in the nervous system due to the narrow expression pattern of p35 and p39 in post-mitotic neurons. A large number of CDK5 substrates have been identified although no substrates have been specifically attributed to p35 or p39. Substrates of CDK5 include p35, PAK1, Src, β-catenin, tau, neurofilament-H, neurofilament-M, synapsin1, APP, DARPP32, PP1-inhibitor and Rb. p35 is rapidly degraded (T1/2 <20 min) by the ubiquitin-proteasome pathway (1). However, p35 stability increases as CDK5 kinase activity decreases, likely as a result of decreased phosphorylation of p35 at Thr138 by CDK5 (2). Proteolytic cleavage of p35 by calpain produces p25 upon neurotoxic insult, resulting in prolonged activation of CDK5 by p25. Accumulation of p25 is found in neurodegenerative diseases such as Alzheimer disease and amyotrophic lateral sclerosis (ALS) (3,4).

1. Dhavan, R. and Tsai, L.H. (2001) Nat Rev Mol Cell Biol 2, 749-59.
2. Patrick, G.N. et al. (1998) J Biol Chem 273, 24057-64.
3. Lee, M.S. et al. (2000) Nature 405, 360-4.
4. Kusakawa, G. et al. (2000) J Biol Chem 275, 17166-72.

注意事项：

Storage: CDK5 siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC.