SignalSilence? AUF1/hnRNP D siRNA I

• 产品编号：CST-12763S      品牌：CST       原厂货号：12763S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence® AUF1/hnRNP D siRNA I能够抑制人，小鼠以及猴子中AUF1/hnRNP D的表达。Cell Signaling Technology （CST）的SignalSilence® AUF1/hnRNP D siRNA I可以通过RNA干扰特异性抑制AUF1/hnRNP D的表达。RNA干扰是一种通过传递双链RNA分子到细胞中从而选择性沉默目的基因表达的方法。所有SignalSilence® siRNA 产品均经过严格的内部测试并通过Western验证可有效降低目的蛋白的表达。通过三苯基分析对寡核苷酸合成进行逐个碱基的监测以保证适当的耦合效率，随后通过亲和固相提取来纯化寡核苷酸。退火的双链RNA需要通过质谱进行进一步的分析从而确定提取物的组成。为了保证批次间一致性最大化，每一批次产品都要通过质谱与前一批次的产品进行比较。CST建议使用100 nM SignalSilence® AUF1/hnRNP D siRNA I进行转染，并在48-72小时后裂解细胞。具体的转染操作步骤请根据转染试剂生产商提供的方法进行。如您在使用过程中有任何问题请随时联系我们。每瓶中含有100次转染所需用量。每次转染siRNA的终浓度为100 nM，以上用量为24孔板中转染，每孔总体积为300 μl。AU-rich element RNA binding protein 1 (AUF1)也称异质核糖核蛋白D (hnRNP D)。AUF1能够结合到目的mRNA的AU富集元件(ARE)上，并调控mRNA降解（1,2）。 它的目的基因非常广泛，包括IL-1, IL-2, IL-3, Myc, TNF-α以及cyclin D1 (2)。AUF1结合到Myc mRNA后同样会影响Myc的翻译（3）。最近的研究有证据表明AUF1还参与了转录调控。AUF1能与许多基因的启动子结合，包括补体受体2（4），脑啡肽（5）以及α-胎蛋白（6）。AUF1还能与端粒酶的催化亚基Tert启动子及富G重复序列结合，从而调控端粒的维持和正常老化（7,8）。AUF1有四个异构体：p37, p40, p42, 以及 p45，它们都是由同一个单一转录本的可变剪接产生的 (9,10)。所有AUF1的异构体都能够在核和胞质间穿梭（11,12）。这些异构体有不同的定位，并可与展现AUF1功能多样性的不同目的mRNA结合（2）。

1. Brewer, G. (1991) Mol Cell Biol 11, 2460-6.
2. Gratacós, F.M. and Brewer, G. (2010) Wiley Interdiscip Rev RNA 1, 457-73.
3. Liao, B. et al. (2007) Nat Struct Mol Biol 14, 511-8.
4. Tolnay, M. et al. (2000) Biochem J 348 Pt 1, 151-8.
5. Dobi, A. et al. (2006) J Biol Chem 281, 28889-900.
6. Jiao, R. et al. (2006) J Cell Biochem 98, 1257-70.
7. Eversole, A. and Maizels, N. (2000) Mol Cell Biol 20, 5425-32.
8. Pont, A.R. et al. (2012) Mol Cell 47, 5-15.
9. Dempsey, L.A. et al. (1998) Genomics 49, 378-84.
10. Wagner, B.J. et al. (1998) Genomics 48, 195-202.
11. Zhang, W. et al. (1993) Mol Cell Biol 13, 7652-65.
12. Sarkar, B. et al. (2003) J Biol Chem 278, 20700-7.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence® AUF1/hnRNP D siRNA I inhibits human, mouse, and monkey AUF1/hnRNP D expression.

Description

SignalSilence® AUF1/hnRNP D siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit AUF1/hnRNP D 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® AUF1/hnRNP D 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

AU-rich element RNA binding protein 1 (AUF1) is also known as heterogeneous ribonucleoprotein D (hnRNP D). AUF1 binds to the AU rich element (ARE) of target mRNA and regulates mRNA decay (1,2). It has a broad range of target genes including IL-1, IL-2, IL-3, Myc, TNF-α, and cyclin D1 (2). Binding of AUF1 to Myc mRNA also affects translation of Myc (3). Recent studies have provided evidence that AUF1 is also involved in the regulation of transcription. AUF1 binds to the promoters of various genes including complement receptor 2 (4), enkephalin (5), and α-fetoprotein (6). AUF1 also binds to the telomerase catalytic subunit Tert promoter and the G-rich telomeric repeat, thus regulating telomere maintenance and normal aging (7,8). AUF1 has four isoforms produced by alternative splicing of a single transcript: p37, p40, p42, and p45 (9,10). All AUF1 isoforms shuttle between the nucleus and cytoplasm (11, 12). These isoforms have distinct localization and bind to different target mRNAs that contribute to the diversity of AUF1 function (2).

1. Brewer, G. (1991) Mol Cell Biol 11, 2460-6.
2. Gratacós, F.M. and Brewer, G. (2010) Wiley Interdiscip Rev RNA 1, 457-73.
3. Liao, B. et al. (2007) Nat Struct Mol Biol 14, 511-8.
4. Tolnay, M. et al. (2000) Biochem J 348 Pt 1, 151-8.
5. Dobi, A. et al. (2006) J Biol Chem 281, 28889-900.
6. Jiao, R. et al. (2006) J Cell Biochem 98, 1257-70.
7. Eversole, A. and Maizels, N. (2000) Mol Cell Biol 20, 5425-32.
8. Pont, A.R. et al. (2012) Mol Cell 47, 5-15.
9. Dempsey, L.A. et al. (1998) Genomics 49, 378-84.
10. Wagner, B.J. et al. (1998) Genomics 48, 195-202.
11. Zhang, W. et al. (1993) Mol Cell Biol 13, 7652-65.
12. Sarkar, B. et al. (2003) J Biol Chem 278, 20700-7.

注意事项：

Storage: AUF1/hnRNP D siRNA I is supplied in RNAse-free
water. Aliquot and store at -20ºC.