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siRNA pools(siPOOLs)

更新时间:2023-05-04

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简要描述:
siRNA pools(siPOOLs)是30种优化设计的高复
杂性siRNA池,经证明可有效消除脱靶效应,并提
高结果的可靠性(Hannus et al., 2014)。Pack Hunter (pooling) 方法通过将当个siRNA的浓度稀释到刺激表型的阈值以下来对抗单个siRNA的脱靶。

 

 

siRNA pools(siPOOLs)

一款高效、特异性的RNAi产品

 

对于靶向基因沉默,RNA干扰(RNAi)具有易于操作、快速结果、高效率、广泛适用于各种细胞类型的多重优 势。其具有瞬转效应和剂量依赖效应,这点与小分子非常相似。然而,目前的siRNA试剂其特异性和基因沉默 效率可变,阻碍了它们作为药物发现和基因研究工具的应用。

siTOOLs Biotech由Michael Hannus博士和Gunter Meister教授于2013年9月成立,由德国雷根斯堡大学 和Intana Bioscience制药服务公司联合运营。目前推出有3个产品系列:(1)siPOOLs:一种siRNA混合物,有 效增加基因沉默特异性,“稀释”传统RNA干扰试剂的脱靶效应。(2)raPOOLs:一种稳健的RNA亲和纯化方 案,适用于基因功能和相互作用的生物化学研究。(3)riboPOOLs:适合任意物种的核糖体RNA去除试剂,高效且经济。


可靠表型的特异性基因沉默工具

siPOOLs是一款经过优化设计的高复杂性的包含30条siRNA的混合物,经证明可有效消除脱靶效应,并提高结果的可靠性(Hannus et al., 2014)。 Pack Hunter (pooling) 方法通过将当个siRNA的 浓度稀释到刺激表型的阈值以下来对抗单个siRNA的脱靶。 借助专有的设计算法,siPOOLs中的siRNA序列经 过优化,以实现最大的转录本覆盖率,高效杂交并 对旁系同源基因进行过滤,从而实现高效和特异性 的基因沉默。


siPOOLs产品优势

使用简单快捷:siPOOLs与多种转染试剂兼容,几天内就能看 到结果。 

高度特异性且有效:siPOOLs在标准细胞系中将脱靶率降低5-25 倍,且在1-3 nM下实现基因敲低率≥70%。 

一致的表型:与siRNA相反,由序列非依赖性的siPOOL产生 的表型高度一致。 

确保经过检验:通过RT-qPCR进行siPOOL验证,如果在最佳 转染下敲低率低于70%,则有可能重新设计。 

使用最新注释进行定制设计:专业的设计,确保优化热力学特性且避免旁系 同源基因。 

HPLC纯化且无毒:所有siPOOL均经过HPLC纯化,可降低污染物 和副作用的风险。


关键问题:siRNAs的脱靶效应

科学家们一直在使用RNA干扰(RNAi)作为研究基因功能的快速有效的工具。然而,短干扰RNA(siRNA)的脱靶效应和可变性能仍然是一个令人头疼的缺点,在验证工作中消耗了宝贵的时间和资源。

638157808712364521244.png

siRNA通常与靶RNA转录本互补结合,通过RNAi机制指导其降解。脱靶效应主要是由siRNA模拟内源性 基因调节因子microRNA(miRNA)引起的。由于miRNA只需要6个碱基种子匹配到3'非翻译区(UTR)即可触 发转录本下调,因此siRNA在通过这种机制处理时可以改变许多意外靶标的表达。


siPOOLs如何提高特异性

单个siRNA或含有3-4个siRNA的低复杂度siRNA库,经常击中多个脱靶基因并表现出易变的靶基因敲低。 siPOOL是高度复杂且确定的30个siRNA池,每个siRNA以皮摩尔工作浓度存在。因此, · 稀释了每个siRNA的脱靶特征,提高了靶向特异性。 · 确保了靶基因的协同敲低,产生更稳健、更可靠的结果。

 

638157808723771273149.png

 

使用siPOOL具有更高的特异性

 

使用siPOOL具有更高的特异性.png

特异性好的试剂应仅影响其靶标基因。 

HeLa细胞中微阵列的表达谱分析显示,单个siRNA可以诱导许多脱靶基因(红点),而针对同一靶基因(绿 点)并包含非特异性siRNA的siPOOL大大降低了脱靶效应。


通过siPOOL实现更好的重现性和有效的敲低效率

 

通过siPOOL实现更好的重现性和有效的敲低效率.png

普通siRNA的敲低效率差异很大。 

与单个siRNA相比,针对同一基因的siPOOLs具有相似的敲低效率,表明具有更高的稳健性和可重复性(左 图和中图:靶RNA水平的实时定量PCR测量的相关图)。 siPOOLs也表现出有效的基因敲低。在常用的细胞系中,许多基因通常在1 nM浓度下实现75-98%的基 因敲低效率(右图)。


您可以信赖的表型

 

您可以信赖的表型.png

 

如果RNAi效果可靠且特异,靶向同一基因的两种试剂应产生相似的表型。 

每个基因分别使用两个siPOOL池,一个来自我们的人类激酶siPOOL文库,一个是市售的每个基因含3条 siRNAs的文库,科学家在A549细胞中筛选了36个基因并检测了细胞活力。仅siPOOLs产生了较好的一致性表型。


siTOOLs生物技术始终致力于帮助学术研究人员、科学家、制药/生物技术公司和RNAi筛选人员。目前,已帮助许多论文 发表在《自然》、《细胞》、《自然医学》、《科学报告》等顶级期刊上。期待与您的合作,我们将为您的RNA研究提供量身定制 的分子工具。


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