SignalSilence? HELLS siRNA I

• 产品编号：CST-11822S      品牌：CST       原厂货号：11822S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNA干扰对照 > RNAi阳性对照
• 应用分类：

描述：

来自Cell Signaling Technology （CST）的SignalSilence® HELLS siRNA I可以通过RNA干扰特异性抑制HELLS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建议使用100 nM SignalSilence® HELLS siRNA II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。

HELLS或LSH1是一种淋巴特异性解旋酶，被认为参与细胞增殖和白血病发生（1，2）。它被认为是一个染色质重塑因子， DNMT1介导的的甲基化维持以及DNMT3A/DNMT3B介导的从头甲基化都需要它。人们认为HELLS在甲基化维持中的作用主要限制在重复DNA序列上包括主要和次要卫星序列，而不是单拷贝基因（3,4）；最近有证据表明其在全基因组非重复序列的半胱氨酸甲基化中起作用（5）。从头甲基化维持与一些已知的参与细胞多能性和谱系决定的特定基因的沉默有关。

1. Lee, D.W. et al. (2000) Cancer Res 60, 3612-22.
2. Fan, T. et al. Epigenetics 3, 134-42.
3. Dennis, K. et al. (2001) Genes Dev 15, 2940-4.
4. Muegge, K. (2005) Biochem Cell Biol 83, 548-54.
5. Tao, Y. et al. (2011) Proc Natl Acad Sci USA 108, 5626-31.
6. Xi, S. et al. (2009) Stem Cells 27, 2691-702.
7. Xi, S. et al. (2007) Proc Natl Acad Sci USA 104, 14366-71.

原厂资料：

Description

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

HELLS, or LSH1, is a lymphoid-specific helicase thought to be involved in cellular proliferation and leukemogenesis (1,2). It is believed to be a chromatin remodeler and is required for DNMT1-mediated methylation maintenance and DNMT3A/DNMT3B-mediated de novo methylation. The role of HELLS in methylation maintenance was thought to be largely confined to repetitive DNA sequences, including major and minor satellite sequences, rather than single copy genes (3,4); recent evidence suggests a role in genome-wide cysteine methylation at non-repeat sequences (5). De novo methylation maintenance is associated with silencing of specific genes, some known to be involved in pluripotency and lineage commitment (6,7).

1. Lee, D.W. et al. (2000) Cancer Res 60, 3612-22.
2. Fan, T. et al. Epigenetics 3, 134-42.
3. Dennis, K. et al. (2001) Genes Dev 15, 2940-4.
4. Muegge, K. (2005) Biochem Cell Biol 83, 548-54.
5. Tao, Y. et al. (2011) Proc Natl Acad Sci USA 108, 5626-31.
6. Xi, S. et al. (2009) Stem Cells 27, 2691-702.
7. Xi, S. et al. (2007) Proc Natl Acad Sci USA 104, 14366-71.

注意事项：

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