SignalSilence? Mitofusin-1 siRNA I

• 产品编号：CST-13274S      品牌：CST       原厂货号：13274S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence® Mitofusin-1 siRNA I能抑制人，小鼠和大鼠mitofusin-1表达。Cell Signaling Technology (CST) 公司的SignalSilence® Mitofusin-1 siRNA I f帮助研究人员使用RNA干扰技术特异性抑制LIMK1表达，该技术通过向细胞内导入双链RNA分子以选择性的抑制某些基因的表达。所有CST公司的SignalSilence® siRNA都经过了严格的内部测试，western分析显示能够能够降低目标蛋白的表达。寡核苷酸合成经过了三苯甲基的逐碱基分析以确保耦合效率。随后经过亲和固相萃取纯化。退火的RNA双链在经过质谱分析以确保双链的准确性。新批次都会经过质谱分析与之前的批次进行比对以保证不同批次之间的稳定性。CST建议转染100 nM SignalSilence® Mitofusin-1 siRNA I，48-72小时后裂解细胞。转染的步骤请参阅转染试剂的说明书。关于使用问题，可以随时咨询CST。 每管试剂能进行100次转染，每次转染量相当于在转染24孔板时，每孔总体积为300μl培养基中siRNA的终浓度为100nM。线粒体融合蛋白是线粒体跨膜GTPases可以调控线粒体融合，出现在线粒体分裂过程中，对于维持结构和遗传的完整性是必须的(1,2)。哺乳动物中已经描述过两个线粒体融合蛋白，mitofusin-1和-2，它们的氨基酸序列有60%相同，并且共同作用以调控线粒体融合(3)。普遍认为线粒体融合对于正常细胞生长和发育是非常重要的(4)，可能演变为一种抵消mtDNA突变有害影响的机制(3)。小鼠体内任一线粒体融合蛋白发生无意突变都会导致胚胎致死，条件性敲除实验证明骨骼肌中同时敲除mitofusin-1和mitofusin-2会导致严重的线粒体功能失调(3)。

1. Zhang, Y. and Chan, D.C. (2007) FEBS Lett 581, 2168-73.
2. Chan, D.C. (2006) Annu Rev Cell Dev Biol 22, 79-99.
3. Chen, H. et al. (2010) Cell 141, 280-9.
4. Bereiter-Hahn, J. and Vöth, M. (1994) Microsc Res Tech 27, 198-219.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence^® Mitofusin-1 siRNA I inhibits human, mouse and rat mitofusin-1 expression.

Mitofusin-1 siRNA II inhibits human and mouse mitofusin-1 expression.

Description

SignalSilence^® Mitofusin-1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit mitofusin-1 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^® Mitofusin-1 siRNA I 48 to 72 hours prior to cell lysis. For transfection procedure, follow the 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

Mitofusins are mitochondrial transmembrane GTPases that function to regulate mitochondrial fusion, a process that occurs in concert with mitochondrial division and is necessary for the maintenance of structural and genetic mitochondrial integrity (1,2). Two mitofusins have been described in mammals, mitofusin-1 and -2, which share 60% amino acid identity and appear to function coordinately to regulate mitochondrial fusion (3). Mitochondrial fusion is widely recognized as important for normal cell growth and development (4), and may have evolved as a mechanism to offset the deleterious effects of mtDNA mutations (3). Null mutations in either mitofusin are embryonic lethal in mice, whereas conditional knockout studies have shown that combined deletion of mitofusin-1 and mitofusin-2 in skeletal muscle results in severe mitochondrial dysfunction (3).

1. Zhang, Y. and Chan, D.C. (2007) FEBS Lett 581, 2168-73.
2. Chan, D.C. (2006) Annu Rev Cell Dev Biol 22, 79-99.
3. Chen, H. et al. (2010) Cell 141, 280-9.
4. Bereiter-Hahn, J. and Vöth, M. (1994) Microsc Res Tech 27, 198-219.

注意事项：

Storage: Mitofusin-1 siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC