SignalSilence? USP1 siRNA I

• 产品编号：CST-6490S      品牌：CST       原厂货号：6490S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence^® USP1 siRNA I能够抑制和猴中USP1的表达。来自Cell Signaling Technology (CST)的SignalSilence^® USP1 siRNA I可以帮助研究者通过RNA干扰特异性地抑制USP1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^® USP1 siRNA II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。每小瓶可供100次转染，每次转染量相当于在转染24孔板时，每孔总体积为300μl培养基中siRNA的终浓度为100nM。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。USP1属于USP亚家族，并在细胞周期中以依赖转录和泛素-蛋白体的机制被调节(3)。USP1是一种核蛋白，并定位于染色质，在染色质中与FANCD2特异性地联合。USP1去泛素化作用单泛素化的FANCD2，在DNA损伤修复和Chk1蛋白稳定中起重要作用(3,4)。USP1的另一重要靶标是PCNA。USP1去泛素化作用单泛素化的PCNA，因此负向调节DNA损伤过程中PCNA-介导的跨损伤DNA复制(TLS) (5)。USP1与含有WD40 重复序列的UAF1的相互作用，导致活化的USP1/UAF1复合物的形成，该复合物是USP1去泛素化作用活性所必须的(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. Nijman, S.M. et al. (2005) Mol Cell 17, 331-9.
4. Guervilly, J.H. et al. (2011) Hum Mol Genet , .
5. Huang, T.T. et al. (2006) Nat Cell Biol 8, 339-47.
6. Cohn, M.A. et al. (2007) Mol Cell 28, 786-97.
7. Cohn, M.A. et al. (2009) J Biol Chem 284, 5343-51.

原厂资料：

Homology

Species predicted to react based on 100% sequence homology: Monkey

Specificity / Sensitivity

SignalSilence® USP1 siRNA I inhibits human and monkey USP1 expression.

Description

SignalSilence® USP1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit USP1 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® USP1 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. 
 Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

Background

Ubiquitinating enzymes (UBEs) catalyze protein ubiquitination, a reversible process countered by deubiquitinating enzymes (DUB) action (1,2). There are five DUB subfamilies including the USP, UCH, OTU, MJD, and JAMM enzymes. USP1 belongs to the USP subfamily and is regulated in a cell cycle dependent manner by both transcriptional and ubiquitin-proteosomal mechanisms (3). USP1 is a nuclear protein and localizes to chromatin where it is specifically associated with FANCD2. USP1 deubiquitinates monoubiquitinated FANCD2, which plays an important role in DNA damage repair and Chk1 protein stability (3,4). Another important target of USP1 is PCNA. USP1 deubiquitinates monoubiquitinated PCNA, thereby negatively regulating PCNA-mediated translesion synthesis (TLS) during DNA repair (5). USP1 interaction with UAF1, a WD40 repeat-containing protein, leads to formation of an activated USP1/UAF1 complex, which is required for the deubiquitinase activity of USP1 (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. Nijman, S.M. et al. (2005) Mol Cell 17, 331-9.
4. Guervilly, J.H. et al. (2011) Hum Mol Genet , .
5. Huang, T.T. et al. (2006) Nat Cell Biol 8, 339-47.
6. Cohn, M.A. et al. (2007) Mol Cell 28, 786-97.
7. Cohn, M.A. et al. (2009) J Biol Chem 284, 5343-51.

注意事项：

Storage: Storage: USP1 siRNA I is supplied in RNAse-free
water. Aliquot and store at -20ºC