SignalSilence? USP14 siRNA I

• 产品编号：CST-7158S      品牌：CST       原厂货号：7158S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

来自Cell Signaling Technology (CST)的SignalSilence^® USP14 siRNA I可以帮助研究者通过RNA干扰特异性地抑制USP14的表达，这种方法可以通过将双链RNA分子传递到细胞内从而使基因表达有选择的沉默。来自CST的所有的SignalSilence^® siRNA产品都是经过内部严格检测的，并且通过Western blot 分析证明确实能够减少目的蛋白的表达。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推荐使用100 nM SignalSilence^® USP14 siRNA I 进行转染，48到72小时后对细胞进行裂解。转染步骤按照转染试剂说明书提供的步骤进行。遇到任何使用方面的问题，请随时联系CST。泛素化作用酶（UBEs）催化蛋白的泛素化，与去泛素化作用酶(DUB)的过程可逆(1,2)。5种DUB亚家族已经被确定，包括USP, UCH, OTU, MJD和JAMM酶。在人类，有3种DUBs蛋白酶体：PSMD14 (POH1/RPN11), UCH37 (UCH-L5)和泛素特异蛋白酶14，其又被认为是tRNA-鸟嘌呤转糖酶(USP14/TGT60 kDa)的60 kDa亚基。USP14通过与19S调节颗粒的PSMD2 (S2/hRPN1)亚基可逆结合，被募集到蛋白酶体周围。尽管PSMD14可以促进底物的降解(3,4)，USP14被认为可以对抗底物的降解(5-8)。尽管这两种DUBs蛋白酶体具有相反作用的机制尚不明确，但USP14在26S蛋白酶体与底物对接时可以将泛素从底物移除。此外，USP14修饰泛素链远端的泛素残基，因此提高该链对蛋白酶体的泛素受体的亲和力，并提高底物的蛋白稳定性(6,9,10)。研究表明USP14调节哺乳动物连接活性中具有生理作用(11)；对小分子抑制剂的这一活性，有助于治疗神经变性疾病和癌症(5,12)。

1. Nijman, S.M. et al. (2005) Cell 123, 773-86.
2. Nalepa, G. et al. (2006) Nat Rev Drug Discov 5, 596-613.
3. Verma, R. et al. (2002) Science 298, 611-5.
4. Yao, T. and Cohen, R.E. (2002) Nature 419, 403-7.
5. Lee, B.H. et al. (2010) Nature 467, 179-84.
6. Lam, Y.A. et al. (1997) Nature 385, 737-40.
7. Koulich, E. et al. (2008) Mol Biol Cell 19, 1072-82.
8. Jacobson, A.D. et al. (2009) J Biol Chem 284, 35485-94.
9. Hanna, J. et al. (2006) Cell 127, 99-111.
10. Thrower, J.S. et al. (2000) EMBO J 19, 94-102.
11. Wilson, S.M. et al. (2002) Nat Genet 32, 420-5.
12. D'Arcy, P. et al. (2011) Nat Med, Epub ahead of print.

原厂资料：

Description

SignalSilence® USP14 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit USP14 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® USP14 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

Ubiquitinating enzymes (UBEs) catalyze protein ubiquitination, a reversible process countered by deubiquitinating enzyme (DUB) action (1,2). Five DUB subfamilies are recognized, including the USP, UCH, OTU, MJD, and JAMM enzymes. In humans, there are three proteasomal DUBs: PSMD14 (POH1/RPN11), UCH37 (UCH-L5), and Ubiquitin-Specific Protease 14, which is also known as the 60 kDa subunit of tRNA-guanine transglycosylase (USP14/TGT60 kDa). USP14 is recruited to the proteasome through its reversible association with the PSMD2 (S2/hRPN1) subunit of the 19S regulatory particle. Whereas PSMD14 appears to promote substrate degradation (3,4), USP14 is thought to antagonize substrate degradation (5-8). While the underlying mechanism for the opposing roles of these two proteasomal DUBs is still uncertain, it is thought that USP14 removes ubiquitin from substrate upon docking of the substrate with the 26S proteasome. Furthermore, USP14 trims ubiquitin residues from the distal end of the polyubiquitin chain, thus decreasing the affinity of the chain for the ubiquitin receptors of the proteasome, and allowing for enhanced substrate stability (6,9,10). Studies have elucidated a physiologic role for USP14 in regulating synaptic activity in mammals (11); targeting this activity with small molecule inhibitors has potential benefits for the treatment of neurodegenerative diseases and cancer (5,12).

1. Nijman, S.M. et al. (2005) Cell 123, 773-86.
2. Nalepa, G. et al. (2006) Nat Rev Drug Discov 5, 596-613.
3. Verma, R. et al. (2002) Science 298, 611-5.
4. Yao, T. and Cohen, R.E. (2002) Nature 419, 403-7.
5. Lee, B.H. et al. (2010) Nature 467, 179-84.
6. Lam, Y.A. et al. (1997) Nature 385, 737-40.
7. Koulich, E. et al. (2008) Mol Biol Cell 19, 1072-82.
8. Jacobson, A.D. et al. (2009) J Biol Chem 284, 35485-94.
9. Hanna, J. et al. (2006) Cell 127, 99-111.
10. Thrower, J.S. et al. (2000) EMBO J 19, 94-102.
11. Wilson, S.M. et al. (2002) Nat Genet 32, 420-5.
12. D'Arcy, P. et al. (2011) Nat Med, Epub ahead of print.

注意事项：

Storage: USP14 siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC.