SignalSilence? ULK1 siRNA I

• 产品编号：CST-7000S      品牌：CST       原厂货号：7000S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence® ULK1 siRNA I能够抑制人和猴的ULK1的表达。来自Cell Signaling Technology (CST)的SignalSilence^® ULK1 siRNA I 可以帮助研究者通过RNA干扰特异性地抑制 ULK1 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^® ULK1 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。

UNC-51样激酶1和2(ULK1, ULK2)，是两种相关丝氨酸/苏氨酸激酶，研究发现是秀丽隐杆线虫基因UNC-51的哺乳动物同源物，它的突变会导致轴突的异常延伸与生长(1-4)。两种蛋白广泛表达，并且在一个中央脯氨酸/丝氨酸富含区域，和一个高度保守的羧基末端区域的后面含有一个氨基末端激酶域。关于ULK1和ULK2在轴突生长中的作用，有关研究表明，这些激酶主要定位于神经生长锥中，并且参与临界生长因子的内吞作用，如NGF(5)。酵母双杂交试验研究发现，ULK1/2与内吞作用途径的调节器有关，如SynGAP 和 syntenin(6)。ULK1/2与酵母自噬蛋白Agt1/Apg1具有结构相似性(7)。使用siRNA的敲除实验证明了，ULK1对自噬是必不可少的，即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(8)。Atg1/ULK1能够作为控制通路的多重信号的聚焦点(11)，并且能够与多种调节磷酸化状态和蛋白运输的自噬相关蛋白(Atg)相结合(12-16)。AMPK，在低营养条件下的活化，可以直接磷酸化ULK1的多个位点，如Ser317、Ser555 和 Ser777 (17,18)。相反，mTOR，能够调节细胞生长和自噬抑制物，可以磷酸化ULK1的Ser757位点，并破坏ULK1和AMPK的相互作用(17)。

1. Ogura, K. et al. (1994) Genes Dev 8, 2389-400.
2. Kuroyanagi, H. et al. (1998) Genomics 51, 76-85.
3. Yan, J. et al. (1998) Biochem Biophys Res Commun 246, 222-7.
4. Yan, J. et al. (1999) Oncogene 18, 5850-9.
5. Zhou, X. et al. (2007) Proc Natl Acad Sci USA 104, 5842-7.
6. Tomoda, T. et al. (2004) Genes Dev 18, 541-58.
7. Matsuura, A. et al. (1997) Gene 192, 245-50.
8. Chan, E.Y. et al. (2007) J Biol Chem 282, 25464-74.
9. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
10. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
11. Stephan, J.S. and Herman, P.K. (2006) Autophagy 2, 146-8.
12. Okazaki, N. et al. (2000) Brain Res Mol Brain Res 85, 1-12.
13. Young, A.R. et al. (2006) J Cell Sci 119, 3888-900.
14. Kamada, Y. et al. (2000) J Cell Biol 150, 1507-13.
15. Lee, S.B. et al. (2007) EMBO Rep 8, 360-5.
16. Hara, T. et al. (2008) J Cell Biol 181, 497-510.
17. Kim, J. et al. (2011) Nat Cell Biol 13, 132-41.
18. Egan, D.F. et al. (2011) Science 331, 456-61.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence® ULK1 siRNA I inhibits human and monkey ULK1 expression.

Description

SignalSilence® ULK1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit ULK1 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® ULK1 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

Two related serine/threonine kinases, UNC-51-like kinase 1 and 2 (ULK1, ULK2), were discovered as mammalian homologs of the C. elegans gene UNC-51 in which mutants exhibited abnormal axonal extension and growth (1-4). Both proteins are widely expressed and contain an amino-terminal kinase domain followed by a central proline/serine rich domain and a highly conserved carboxy-terminal domain. The roles of ULK1 and ULK2 in axon growth have been linked to studies showing that the kinases are localized to neuronal growth cones and are involved in endocytosis of critical growth factors, such as NGF (5). Yeast two-hybrid studies found ULK1/2 associated with modulators of the endocytic pathway, SynGap and syntenin (6). Structural similarity of ULK1/2 has also been recognized with the yeast autophagy protein Atg1/Apg1 (7). Knockdown experiments using siRNA demonstrated that ULK1 is essential for autophagy (8), a catabolic process for the degradation of bulk cytoplasmic contents (9,10). It appears that Atg1/ULK1 can act as a convergence point for multiple signals that control autophagy (11), and can bind to several autophagy-related (Atg) proteins, regulating phosphorylation states and protein trafficking (12-16).AMPK, activated during low nutrient conditions, directly phosphorylates ULK1 at multiple sites including Ser317, Ser555, and Ser777 (17,18). Conversely, mTOR, which is a regulator of cell growth and an inhibitor of autophagy, phosphorylates ULK1 at Ser757 and disrupts the interaction between ULK1 and AMPK (17).

1. Ogura, K. et al. (1994) Genes Dev 8, 2389-400.
2. Kuroyanagi, H. et al. (1998) Genomics 51, 76-85.
3. Yan, J. et al. (1998) Biochem Biophys Res Commun 246, 222-7.
4. Yan, J. et al. (1999) Oncogene 18, 5850-9.
5. Zhou, X. et al. (2007) Proc Natl Acad Sci USA 104, 5842-7.
6. Tomoda, T. et al. (2004) Genes Dev 18, 541-58.
7. Matsuura, A. et al. (1997) Gene 192, 245-50.
8. Chan, E.Y. et al. (2007) J Biol Chem 282, 25464-74.
9. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
10. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
11. Stephan, J.S. and Herman, P.K. (2006) Autophagy 2, 146-8.
12. Okazaki, N. et al. (2000) Brain Res Mol Brain Res 85, 1-12.
13. Young, A.R. et al. (2006) J Cell Sci 119, 3888-900.
14. Kamada, Y. et al. (2000) J Cell Biol 150, 1507-13.
15. Lee, S.B. et al. (2007) EMBO Rep 8, 360-5.
16. Hara, T. et al. (2008) J Cell Biol 181, 497-510.
17. Kim, J. et al. (2011) Nat Cell Biol 13, 132-41.
18. Egan, D.F. et al. (2011) Science 331, 456-61.

注意事项：

Storage: ULK1 siRNA I is supplied in RNAse-free water. Aliquot
and store at -20ºC.