SignalSilence? RMP siRNA II

• 产品编号：CST-6495S      品牌：CST       原厂货号：6495S
• 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)公司的SignalSilence® RMP siRNA II允许研究者通过RNA干扰去特异性抑制RMP蛋白表达，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公司推荐在细胞裂解前48到72小时转染100 nM RMP siRNA II。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。RMP (RPB5介导的蛋白)又称为URI (Unconventional prefoldin RBP5 Interactor)，其被描述作为伴侣蛋白prefoldin (PFD)家族的一个非传统成员，其涉及actin和tubulin蛋白的折叠 (1-4)。像PFDs的α类传统成员一样，RMP蛋白含有通过两个β发夹结构连接的N和C端的α-螺旋型卷曲螺旋结构。此外，RMP蛋白拥有一个RPB5蛋白结合部分和一个长的羧基末端酸性部分。研究指出RMP作为一个人类细胞大分子复合物的组成成分，且作为一个分子支架来组装PFD复合物，这个复合物包含其它PFDs 和转录及泛素化功能相关的蛋白。的确，研究证明RMP通过结合到TFIIF (5)和RBP5 (6)负调控RNA polymerase II依赖的转录，并且通过协调与基因表达有关的营养成分的供给从而涉及mTOR 信号通路(1)。与营养成分供给相关的基因表达的协调能力相符合，RMP被证实是一个S6K1的线粒体底物。S6K1介导RMP在Ser371 位点的磷酸化可引起一系列生物化学反应，反应可构成一个负反馈环，某种程度上，针对抑制S6K1存活信号和确保线粒体凋亡的阈值应与营养和生长因子的供给相符合(7)。

1. Gstaiger, M. et al. (2003) Science 302, 1208-12.
2. Vainberg, I.E. et al. (1998) Cell 93, 863-73.
3. Martín-Benito, J. et al. (2002) EMBO J 21, 6377-86.
4. Geissler, S. et al. (1998) EMBO J 17, 952-66.
5. Wei, W. et al. (2003) Cell Res 13, 111-20.
6. Dorjsuren, D. et al. (1998) Mol Cell Biol 18, 7546-55.
7. Djouder, N. et al. (2007) Mol Cell 28, 28-40.

原厂资料：

Description

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

RMP (RPB5-Mediating Protein), also known as URI (Unconventional prefoldin RBP5 Interactor), was described as an unconventional member of the prefoldin (PFD) family of chaperones that are involved in actin and tubulin folding (1-4). Like conventional members of the α-class of PFDs, RMP contains N- and C-terminal α-helical coiled-coil structures connected by two β hairpins. In addition, RMP possesses an RPB5-binding segment and a long C-terminal acidic segment. It is posited that RMP exists as a component of a macromolecular complex within human cells and functions as a molecular scaffold to assemble a PFD complex containing other PFDs and proteins with functions in transcription and ubiquitination. Indeed, evidence is provided that RMP negatively modulates RNA polymerase II-dependent transcription by binding to TFIIF (5) and RBP5 (6) and is involved in mTOR signaling by coordinating the regulation of nutrient availability with gene expression (1). In accord with its ability to coordinate gene expression with nutrient availability, RMP was shown to be a mitochondrial substrate of S6K1. S6K1-mediated phosphorylation of RMP at Ser371 triggers a series of biochemical events that constitute a negative feedback loop, in part, aimed at restraining S6K1 survival signaling and ensuring that the mitochondrial threshold for apoptosis corresponds to availability of nutrients and growth factors (7).

1. Gstaiger, M. et al. (2003) Science 302, 1208-12.
2. Vainberg, I.E. et al. (1998) Cell 93, 863-73.
3. Martín-Benito, J. et al. (2002) EMBO J 21, 6377-86.
4. Geissler, S. et al. (1998) EMBO J 17, 952-66.
5. Wei, W. et al. (2003) Cell Res 13, 111-20.
6. Dorjsuren, D. et al. (1998) Mol Cell Biol 18, 7546-55.
7. Djouder, N. et al. (2007) Mol Cell 28, 28-40.

注意事项：

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