SignalSilence? Atg14 siRNA I

• 产品编号：CST-6286S      品牌：CST       原厂货号：6286S
• 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^® Atg14 siRNA I 可以帮助研究者通过RNA干扰特异性地抑制Atg14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^® Atg14 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。自噬是自噬溶酶体降解其所包裹的细胞质内容物的一个分解代谢过程。通常情况下，自噬在营养匮乏的时候被激活，但它还与许多生理过程相关，包括生长、分化、神经退行性疾病、感染和癌症。自噬的分子机制主要在酵母中发现的，并称之为自噬相关基因。这些蛋白与自噬体形成有关，自噬体就是被送往溶酶体进行降解的胞浆空泡。III型磷酸肌醇3激酶（PI3K）Vps34调节空泡的运输和自噬（4,5）。许多蛋白包括p105/Vsp15，Beclin-1，UVRAG，Atg14和Rubicon与Vsp34一起决定Vsp34的功能（6-12）。Atg14和Rubicon能够与Beclin-1结合，并且参与具有相对功能的独特复合物的形成（9-12）。Rubicon主要集中于内涵体和溶酶体，能够抑制Vps34脂质激酶活性；Rubicon的下调可以增强自噬和内吞作用（11,12）。相反，Atg14集中于自噬体，隔离膜和内质网，能够增强Vps34活性；Atg14的下调可以抑制饥饿诱导的自噬(11,12)。

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. Levine, B. and Yuan, J. (2005) J Clin Invest 115, 2679-88.
4. Corvera, S. (2001) Traffic 2, 859-66.
5. Yan, Y. and Backer, J.M. (2007) Biochem Soc Trans 35, 239-41.
6. Stack, J.H. et al. (1995) J Cell Biol 129, 321-34.
7. Zeng, X. et al. (2006) J Cell Sci 119, 259-70.
8. Liang, C. et al. (2006) Nat Cell Biol 8, 688-99.
9. Itakura, E. et al. (2008) Mol Biol Cell 19, 5360-72.
10. Sun, Q. et al. (2008) Proc Natl Acad Sci U S A 105, 19211-6.
11. Zhong, Y. et al. (2009) Nat Cell Biol 11, 468-76.
12. Matsunaga, K. et al. (2009) Nat Cell Biol 11, 385-96.

原厂资料：

Description

SignalSilence® Atg14 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Atg14 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 Atg14 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

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). Autophagy is generally activated by conditions of nutrient deprivation but is also 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 is directed by a number of autophagy-related (Atg) genes. These proteins are involved in the formation of autophagosomes, cytoplasmic vacuoles that are delivered to lysosomes for degradation. The class III type phosphoinositide 3-kinase (PI3K) Vps34 regulates vacuolar trafficking and autophagy (4,5). Multiple proteins associate with Vsp34, including p105/Vsp15, Beclin-1, UVRAG, Atg14, and Rubicon, to determine Vsp34 function (6-12). Atg14 and Rubicon were identified based on their ability to bind to Beclin-1 and participate in unique complexes with opposing functions (9-12). Rubicon, which localizes to the endosome and lysosome, inhibits Vps34 lipid kinase activity; knockdown of Rubicon enhances autophagy and endocytic trafficking (11,12). In contrast, Atg14 localizes to autophagosomes, isolation membranes and ER, and can enhance Vps34 activity. Knockdown of Atg14 inhibits starvation-induced autophagy (11,12).

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. Levine, B. and Yuan, J. (2005) J Clin Invest 115, 2679-88.
4. Corvera, S. (2001) Traffic 2, 859-66.
5. Yan, Y. and Backer, J.M. (2007) Biochem Soc Trans 35, 239-41.
6. Stack, J.H. et al. (1995) J Cell Biol 129, 321-34.
7. Zeng, X. et al. (2006) J Cell Sci 119, 259-70.
8. Liang, C. et al. (2006) Nat Cell Biol 8, 688-99.
9. Itakura, E. et al. (2008) Mol Biol Cell 19, 5360-72.
10. Sun, Q. et al. (2008) Proc Natl Acad Sci U S A 105, 19211-6.
11. Zhong, Y. et al. (2009) Nat Cell Biol 11, 468-76.
12. Matsunaga, K. et al. (2009) Nat Cell Biol 11, 385-96.

注意事项：

Storage: Atg14 siRNA I is supplied in RNAse-free water. Aliquot
and store at -20ºC.