SignalSilence? SQSTM1/p62 siRNA I

• 产品编号：CST-6394S      品牌：CST       原厂货号：6394S
• 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^® SQSTM1/p62 siRNA I 可以帮助研究者通过RNA干扰特异性地抑制 SQSTM1/p62 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^® SQSTM1/p62 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。Sequestosome 1 (SQSTM1, p62) 是一种涉及细胞信号、氧化应激和自噬的泛素结合蛋白(1-4)。初次被鉴定为能够结合p56Lck的SH2域的一种蛋白(5)，并且可以独立地与PKCζ相互作用(6,7)。随后研究发现，其能够与泛素相互作用，为多种信号蛋白提供支架，通过蛋白酶体或溶酶体触发蛋白的降解(8)。SQSTM1与TRAF6的相互作用，导致K63连接的TRAF6的多泛素化作用，以及随后的NF-κB通路的激活(9)。蛋白通过SQSTM1形成聚合物，能够被自噬小体降解(4,10,11)。SQSTM1能够结合自噬小体膜蛋白LC3/Atg8，将含有SQSTM-1的蛋白聚合物运输到自噬小体处(12)。在自噬过程中，溶酶体降解自噬小体从而导致SQSTM1水平的降低，相反，自噬抑制剂可以稳定SQSTM1的水平。研究证明了，SQSTM1和氧化应激之间的关系。SQSTM1可以与KEAP1相互作用，KEAP1是NRF2的细胞质抑制剂，一个关键的细胞转录因子(3)。因此，SQSTM1的累积能够导致NRF2的活化。

1. Kirkin, V. et al. (2009) Mol Cell 34, 259-69.
2. Seibenhener, M.L. et al. (2007) FEBS Lett 581, 175-9.
3. Komatsu, M. et al. (2010) Nat Cell Biol 12, 213-23.
4. Bjørkøy, G. et al. (2006) Autophagy 2, 138-9.
5. Joung, I. et al. (1996) Proc Natl Acad Sci USA 93, 5991-5.
6. Sanchez, P. et al. (1998) Mol Cell Biol 18, 3069-80.
7. Puls, A. et al. (1997) Proc Natl Acad Sci USA 94, 6191-6.
8. Vadlamudi, R.K. et al. (1996) J Biol Chem 271, 20235-7.
9. Wooten, M.W. et al. (2005) J Biol Chem 280, 35625-9.
10. Bjørkøy, G. et al. (2005) J Cell Biol 171, 603-14.
11. Komatsu, M. et al. (2007) Cell 131, 1149-63.
12. Pankiv, S. et al. (2007) J Biol Chem 282, 24131-45.

原厂资料：

Description

SignalSilence® SQSTM1/p62 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit SQSTM1/p62 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 SQSTM1/p62 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

Sequestosome 1 (SQSTM1, p62) is a ubiquitin binding protein involved in cell signaling, oxidative stress, and autophagy (1-4). It was first identified as a protein that binds to the SH2 domain of p56Lck (5), and independently found to interact with PKCζ (6,7). It was subsequently found to interact with ubiquitin, providing a scaffold for several signaling proteins and triggering degradation of proteins through the proteasome or lysosome (8). Interaction between SQSTM1 and TRAF6 leads to the K63-linked polyubiquitination of TRAF6 and subsequent activation of the NF-κB pathway (9). Protein aggregates formed by SQSTM1 can be degraded by the autophagosome (4,10,11). SQSTM1 binds autophagosomal membrane protein LC3/Atg8, bringing SQSTM1-containing protein aggregates to the autophagosome (12). Lysosomal degradation of autophagosomes leads to a decrease in SQSTM1 levels during autophagy; conversely, autophagy inhibitors stabilize SQSTM1 levels. Studies have demonstrated a link between SQSTM1 and oxidative stress. SQSTM1 interacts with KEAP1, which is a cytoplasmic inhibitor of NRF2, a key transcription factor involved in cellular responses to oxidative stress (3). Thus, accumulation of SQSTM1 can lead to an increase in NRF2 activity.

1. Kirkin, V. et al. (2009) Mol Cell 34, 259-69.
2. Seibenhener, M.L. et al. (2007) FEBS Lett 581, 175-9.
3. Komatsu, M. et al. (2010) Nat Cell Biol 12, 213-23.
4. Bjørkøy, G. et al. (2006) Autophagy 2, 138-9.
5. Joung, I. et al. (1996) Proc Natl Acad Sci USA 93, 5991-5.
6. Sanchez, P. et al. (1998) Mol Cell Biol 18, 3069-80.
7. Puls, A. et al. (1997) Proc Natl Acad Sci USA 94, 6191-6.
8. Vadlamudi, R.K. et al. (1996) J Biol Chem 271, 20235-7.
9. Wooten, M.W. et al. (2005) J Biol Chem 280, 35625-9.
10. Bjørkøy, G. et al. (2005) J Cell Biol 171, 603-14.
11. Komatsu, M. et al. (2007) Cell 131, 1149-63.
12. Pankiv, S. et al. (2007) J Biol Chem 282, 24131-45.

注意事项：

Storage: SQSTM1/p62 siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC