SignalSilence? USP10 siRNA I

• 产品编号：CST-7747S      品牌：CST       原厂货号：7747S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

Cell Signaling Technology (CST)公司生产的SignalSilence USP10 siRNA I使得研究者可以使用RNA干扰技术特异性的抑制USP10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公司推荐您在细胞裂解前48-72小时用100nM SignalSilence USP10 siRNA I进行转染。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。泛素化酶(UBEs)催化蛋白泛素化，该过程可被去泛素化酶（DUB）的作用逆转（1,2）。五个DUB亚家族已经被发现，包括USP、UCH、OTU、MJD和JAMM酶。USP10具有和半胱氨酸和组氨酸盒的一样的氨基酸序列，而这一序列是USP去泛素化酶家族的标志性特征。在翻译后水平，USP10似乎可以通过蛋白间相互作用或磷酸化来调节。USP10和Ras-GAP SH3区域结合蛋白（G3BP）的相互作用被发现的确可以抑制其催化泛素链解聚的能力（3）。另外，ATM介导的USP10在Thr42和Ser337的磷酸化被显示出有促进USP10稳定性和促使其在细胞质和细胞浆中重新分配的作用，这涉及到p53的去泛素化、稳定性和遗传毒性应激反应（4）。最近研究显示USP10与USP13和Vps34复合物协调共同发挥作用。USP10与USP13似乎一同对Vps34复合物进行去泛素化，调节这个III类PI3K。Beclin-1是这一复合物的另一组份，其功能是调节USP13的稳定性，而USP13可以去泛素化USP10和稳定其水平。因此Beclin-1可以通过控制USP10的DUB活性间接调节p53稳定性（5）。USP10也在内体区域发挥作用，研究显示其可去泛素化CFTR，从而促进其的内吞回收以及在细胞表面的表达（6,7）。

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. Soncini, C. et al. (2001) Oncogene 20, 3869-79.
4. Yuan, J. et al. (2010) Cell 140, 384-96.
5. Liu, J. et al. (2011) Cell 147, 223-34.
6. Bomberger, J.M. et al. (2009) J Biol Chem 284, 18778-89.
7. Bomberger, J.M. et al. (2011) Channels (Austin) 4, 150-4.

原厂资料：

Description

SignalSilence® USP10 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit USP10 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® USP10 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. USP10 possesses amino acid sequences that match the consensus cysteine and histidine boxes representative of the USP family of deubiquitinating enzymes. At the posttranslational level, USP10 appears to be regulated through both protein-protein interactions and phosphorylation. Indeed, interaction of USP10 with Ras-GAP SH3 domain binding protein (G3BP) has been found to inhibit its ability to catalyze the disassembly of ubiquitin chains (3). Furthermore, ATM-mediated phosphorylation of USP10 at Thr42 and Ser337 was shown to promote USP10 stabilization and redistribution from the cytoplasm to the nucleus, where it functions in p53 deubiquitination, stabilization, and activation in response to genotoxic stress (4). Recently, it was shown that USP10 works in concert with USP13 and Vps34 complexes. USP10, along with USP13, appears to deubiquitinate Vps34 complexes to regulate the levels of this class III PI3K. Beclin-1, another component of these complexes, functions to regulate the stability of USP13, which can deubiquitinate and stabilize the levels of USP10. Therefore, Beclin-1, can indirectly regulate p53 stability by controlling the DUB activity of USP10 (5). USP10 also functions in the endosomal compartment, where it has been shown to deubiquitinate CFTR in order to enhance its endocytic recycling and cell surface expression (6,7).

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. Soncini, C. et al. (2001) Oncogene 20, 3869-79.
4. Yuan, J. et al. (2010) Cell 140, 384-96.
5. Liu, J. et al. (2011) Cell 147, 223-34.
6. Bomberger, J.M. et al. (2009) J Biol Chem 284, 18778-89.
7. Bomberger, J.M. et al. (2011) Channels (Austin) 4, 150-4.

注意事项：

Storage: USP10 siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC.