灵活经济的RNAi筛选工具
  • 按需灵活选择siRNA、规格和孔板排列
  • 花费更少,筛查更多的靶基因
  • 在GeneGlobe可方便的查找和选择siRNA,并自定义孔板排列方式
  • 创新的siRNA设计最大程度减少脱靶效应
  • 供货速度快,加快实验进程

FlexiPlate siRNA是高度灵活的RNAi筛选工具。根据靶基因可选择每孔0.1 nmol、0.25 nmol或1 nmol的96孔板规格,以及每孔0.1 nmol或0.25 nmol的384孔板规格。可在GeneGlobe选择siRNA,并定义孔板排列方式,十分灵活便利。并可得到很多基因家族的siRNA列表。siRNA通过HP OnGuard siRNA Design设计,该方法结合神经网络技术,专有的同源性分析技术以及多项先进功能,例如3' UTR/seed region分析、序列不对称性分析、SNP位点屏蔽、干扰素序列屏蔽等。

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FlexiPlate siRNA, 0.1 nmol
Custom siRNA set in 96-well plate format, 0.1 nmol
1027411 Varies
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FlexiPlate siRNA, 0.25 nmol
Custom siRNA set in 96-well plate format, 0.25 nmol
1027412 Varies
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FlexiPlate siRNA, 1 nmol
Custom siRNA set in 96-well plate format, 1 nmol
1027413 Varies
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FlexiPlate siRNA (384), 0.1 nmol
Custom siRNA set in 384-well plate format, 0.1 nmol
1027421 Varies
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FlexiPlate siRNA (384), 0.25 nmol
Custom siRNA set in 384-well plate format, 0.25 nmol
1027422 Varies
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The FlexiPlate siRNA is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Performance
先进的siRNA设计

QIAGEN在siRNA设计中的进步性,确保其高度创新且精良的HP OnGuard siRNA Design,能提供有效特异的siRNA。根据来自RNAi试验的数据建立极大容量的数据集,并在此基础上利用神经网络技术进行siRNAs设计,然后使用先进的、非冗余序列数据库和同源分析工具,检查该序列与同一基因组内其他序列的同源性。HP OnGuard siRNA Design具有许多独特先进的特性(见下表)。

HP OnGuard siRNA Design特性
特性描述 参考
神经网络技术 使用BioPredsi神经网络设计siRNA,依托于极大的RNAi数据库。 1-3
全球最大的siRNA验证项目 该项目提供的数据使设计过程更完善,QIAGEN科学家已通过成千上万的siRNA确认其效能。该项目证实大量药用基因组siRNA功能至少有70%可被抑制。 4
同源性分析 该分析使用专用工具以及最新的,非冗余序列数据库。
Affymetrix基因芯片分析 基因组范围siRNA设计的创新性发展最大程度减少脱靶效应。
最新的siRNA靶序列 NCBI数据库的最新数据确保设计的准确性。
不对称性 对siRNA的设计基于5’端碱基对的不等稳定性,使得有义链被降解的同时,反义链在5’端的结合度降低而进入RISC。利用不对称性可制备高度可用的siRNA,且大大降低了错误链进入RISC而导致脱靶效应的风险。 5,6
3' UTR/seed区域分析 使用智能加权分析,多参数搜索与非目的靶标mRNA3’非编码区相匹配的siRNA反义链的源区域。 7-12
SNP位点屏蔽 采用RefSNP数据库排除单核苷酸多态(SNP)的siRNA,由于这种siRNA有效性不同从而提高了siRNA的效果。
干扰素序列屏蔽 屏蔽掉可导致干扰素反应的多重序列基序siRNA,弃用含有这种基序的siRNA。 13,14
1. Huesken, D. et al. (2005) Design of a genome-wide siRNA library using an artificial neural network. Nat. Biotechnol. 23, 995.
2. Mukherji, M. et al. (2006) Genome-wide functional analysis of human cell-cycle regulators. Proc. Natl. Acad. Sci. 103, 14819.
3. Matveeva, O. et al. (2007) Comparison of approaches for rational siRNA design leading to a new efficient and transparent method. Nucleic Acids Res. 35, e63.
4. Krueger, U. et al. (2007) Insights into effective RNAi gained from large-scale siRNA validation screening. Oligonucleotides 17, 237.
5. Aza-Blanc, P. et al. (2003) Identification of modulators of TRAIL-induced apoptosis via RNAi-based phenotypic screening. Mol. Cell 12, 627.
6. Schwarz, D.S. et al. (2003) Asymmetry in the assembly of the RNAi enzyme complex. Cell 115, 199.
7. Farh, K.K. et al. (2005) The widespread impact of mammalian microRNAs on mRNA repression and evolution. Science 310, 1817.
8. Grimson, A. et al. (2007) MicroRNA targeting specificity in mammals: determinants beyond seed pairing. Mol. Cell 27, 91.
9. Jackson, A.L. et al. (2003) Expression profiling reveals off-target gene regulation by RNAi. Nat. Biotechnol. 21, 635.
10. Lewis, B.P., Burge, C.B., and Bartel, D.P. (2005) Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120, 15.
11. Lim, L.P. et al. (2005) Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAs. Nature 433, 769.
12. Saxena, S., Jónsson, Z.O., and Dutta, A. (2003) Small RNAs with imperfect match to endogenous mRNA repress translation. Implications for off-target activity of small inhibitory RNA in mammalian cells. J. Biol. Chem. 278, 44312.
13. Judge, A.D., Sood, V., Shaw, J.R., Fang, D., McClintock, K., and MacLachlan, I. (2005) Sequence-dependent stimulation of the mammalian innate immune response by synthetic siRNA. Nat Biotechnol. 23, 457.
14. Hornung, V. et al. (2005) Sequence-specific potent induction of IFN-alpha by short interfering RNA in plasmacytoid dendritic cells through TLR7. Nat Med. 11, 263.
3' UTR/seed区域分析

研究表明siRNA反义链源区域与非目的靶标mRNA3’非编码区的匹配(见表),可能导致脱靶效应。siRNA双链的反义链上的2-7位是源区域,包括6个碱基。由于siRNA可模拟miRNA的作用,类似的匹配会促进非靶标mRNA的减少。使用QIAGEN设计的siRNA,与源自miRNA,siRNA和鼠RefSeq数据库的3’非编码区专利产品相配合,可对3’UTR/seed区进行分析研究。将每个siRNA与这些序列比对,检查任何可能导致miRNA类脱靶效应的同源序列。

siRNA源区域所有6个碱基与无关靶标3’非编码区序列完全匹配是常见情况,且无需除去这种siRNA。更罕见的是siRNA序列中出现源区域与10个或更多碱基同源匹配的情况,这种同源性很有可能导致脱靶效应,而这些siRNA可能因为与非靶标基因表现出更高的同源性而被弃用。

对于一些靶位点,很难选择到完全非同源性的siRNA。对于这种情况,非相关基因的EntrezGene ID可能是GeneGlobe提供的siRNA的非靶标基因。对这种类型同源性的关注并不意味这些基因一定会受到siRNA的影响,然而如有必要,后续分析中可以认定上述基因为潜在的非靶标基因。

Principle

FlexiPlate siRNA通过筛查实验设计RNAi,以满足特定需求。可按需选择用于阳性和阴性对照的用于任意人类或鼠基因的siRNA使用FlexiPlate siRNA可设计RNAi筛选实验,满足不同需求。可选择任何人类或小鼠基因的siRNA以及所需的阳性或阴性对照。网页界面友好,可任意排列siRNA和其对照在96孔板或384孔板中的位置,或使用多种预设的排列方式设置孔板的位置。siRNA具有0.1 nmol、0.25 nmol或1 nmol的规格以供订购;可按需进行小量或大量siRNA筛查,节省开支。96孔板的规格包括0.1 nmol、0.25 nmol或1 nmol。384孔板的规格包括0.1 nmol或0.25 nmol。

Procedure

使用GeneGlobe数据库可方便的搜索您感兴趣的siRNA,并根据筛查实验设置孔板布局。可上传基因名称、Entrez Gene ID、RefSeq ID、siRNA名称或货号,快速方便实现孔板的订购,可立即订购或保存起以备后用。板式可下载到本地以供记录。

Applications

FlexiPlate siRNA适用于多种RNAi应用:

  • 通路分析
  • 大规模RNAi筛选的后续实验
Feature
Specifications
Design Predesigned/HiPerformance siRNA Design Algorithm
Format Plate
Guarantee/validation No guarantee
Modification No
Scale or yield 0.1 nmol, 0.25 nmol, 1 nmol
Species Human, mouse
Target sequence provided Yes

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Kit Handbooks
1
For transfection of eukaryotic cells with siRNA and miRNA
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