SignalSilence? UVRAG siRNA I

• 产品编号：CST-13219S      品牌：CST       原厂货号：13219S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence^® UVRAG siRNA I 能够抑制人和猴源的UVRAG的表达。来自Cell Signaling Technology (CST)的SignalSilence^® UVRAG siRNA I 可以帮助研究者通过RNA干扰特异性地抑制UVRAG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^® UVARG 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。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。

自噬是自噬溶酶体对其包裹的细胞质内容物进行降解的一种分解代谢过程(1,2)。自噬过程一般在营养匮乏的条件下被激活，但也与一些生理过程有关，如发育、分化、神经变性、感染和癌症(3)。关于自噬的分子机制，已在酵母中被大量发现，并且还提到了如自噬相关基因（Atg）。这些蛋白涉及称为自噬小体的细胞质液泡的形成，并且将其运送到溶酶体处降解。

III型磷酸肌醇3-激酶(PI3KC3)/Vps34调节空泡的运输和自噬(4,5)。多种蛋白与Vsp34有关，包括：p105/Vsp15、Beclin-1、UVRAG、Atg14 和 Rubicon，这些蛋白能够决定Vsp34的功能(6-11)。UVRAG(紫外线辐射抗性相关基因)与Beclin-1/PI3KC3复合物相关，并提高PI3KC3酶活性，及促进自噬，抑制增殖(11)。Beclin-1结合UVRAG可以促进自噬小体的成熟和内吞小泡的运输(12)。同样地，UVRAG存在于肠癌和胃癌中，可以作为一个伴有移码突变的潜在的肿瘤抑制蛋白(13,14)。

1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
3. Corvera, S. (2001) Traffic 2, 859-66.
4. Stack, J.H. et al. (1995) J Cell Biol 129, 321-34.
5. Liang, C. et al. (2008) Nat Cell Biol 10, 776-87.
6. Matsunaga, K. et al. (2009) Nat Cell Biol 11, 385-96.
7. Zhong, Y. et al. (2009) Nat Cell Biol 11, 468-76.
8. Sun, Q. et al. (2008) Proc Natl Acad Sci U S A 105, 19211-6.
9. Itakura, E. et al. (2008) Mol Biol Cell 19, 5360-72.
10. Liang, C. et al. (2006) Nat Cell Biol 8, 688-99.
11. Ionov, Y. et al. (2004) Oncogene 23, 639-45.
12. Kim, M.S. et al. (2008) Hum Pathol 39, 1059-63.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence^® UVRAG siRNA I inhibits human and monkey UVRAG expression.

Description

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

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). It is generally activated by conditions of nutrient deprivation but has also been associated with a number of physiological processes including development, differentiation, neurodegeneration, infection and cancer (3). The molecular machinery of autophagy was largely discovered in yeast and referred to as autophagy-related (Atg) genes. These proteins are involved in the formation of cytoplasmic vacuoles called autophagosomes that are delivered to lysosomes for degradation.

The class III type phosphoinositide 3-kinase (PI3KC3)/Vps34 regulates vacuolar trafficking as well as autophagy (4,5). Multiple proteins have been shown to be associated with Vsp34, including: p105/Vsp15, Beclin-1, UVRAG, Atg14, and Rubicon, which can determine Vsp34 function (6-11). UVRAG (UV radiation resistance-associated gene) is associated with the Beclin-1/PI3KC3 complex and promotes PI3KC3 enzymatic activity and autophagy, while suppressing proliferation (11). Beclin-1 binding to UVRAG promotes both autophagosome maturation and endocytic trafficking (12). UVRAG is also a potential tumor suppressor protein with frameshift mutations observed in colon and gastric carcinomas (13,14).

1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
3. Corvera, S. (2001) Traffic 2, 859-66.
4. Stack, J.H. et al. (1995) J Cell Biol 129, 321-34.
5. Liang, C. et al. (2008) Nat Cell Biol 10, 776-87.
6. Matsunaga, K. et al. (2009) Nat Cell Biol 11, 385-96.
7. Zhong, Y. et al. (2009) Nat Cell Biol 11, 468-76.
8. Sun, Q. et al. (2008) Proc Natl Acad Sci U S A 105, 19211-6.
9. Itakura, E. et al. (2008) Mol Biol Cell 19, 5360-72.
10. Liang, C. et al. (2006) Nat Cell Biol 8, 688-99.
11. Ionov, Y. et al. (2004) Oncogene 23, 639-45.
12. Kim, M.S. et al. (2008) Hum Pathol 39, 1059-63.

注意事项：

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