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查看更多产品信息 Silencer™ GAPDH Positive Control siRNA, in vivo ready - FAQs (4404025)
28 个常见问题解答
我们提供下列体内siRNA对照：
•Ambion In Vivo阴性对照 #1 siRNA（货号4457287, 5nmol; 4459405, 50 nmol; 4457289, 250 nmol）
•Ambion In Vivo GAPDH阳性对照siRNA（货号4457288, 5 nmol; 4459407, 50 nmol, 4457291, 250 nmol）
•Ambion In Vivo Factor VII阳性对照siRNA（货号4459408, 50 nmol; 4457292, 250 nmol）
•Silencer Select阴性对照1号 siRNA, in vivo ready（货号4404020, 250 nmol）
•Silencer GAPDH阳性对照siRNA, in vivo ready（货号4404025, 250 nmol）
Stealth siRNA比非修饰siRNA在血清中稳定的多。这一高稳定性对于体内实验尤其有好处，因为在体内RNAi分子会暴露于更多蛋白酶。通过与 Intradigm合作，Stealth RNAi进行瘤内注射时已被证明具有体内活性。
用于体内实验的Stealth siRNA双链可以使用BLOCK-iT RNAi Express订购，而Ambion in vivo siRNA可以使用此工具（https://www.thermofisher.com/us/en/home/life-science/rnai/introduction-to-in-vivo-rnai/ambion-in-vivo-sirna.html?icid=cvc-invivo-sirna-c3t1）订购。
推荐进行内毒素检测以减少引发免疫反应的风险。
请联系rnairesearcher@invitrogen.com咨询大规模体内siRNA订购或填写询价表格（ www.thermofisher.com/us/en/home/global/forms/custom-sirna-mirna-quote-request.html）。
有数种不同的方法可以用于siRNA导入，包括多种局部导入技术和系统导入技术。我们提供一种脂质体试剂Invivofectamine 3.0用于体内siRNA系统导入。点击此处（https://www.thermofisher.com/us/en/home/life-science/rnai/introduction-to-in-vivo-rnai/invivofectamine-reagent/invivofectamine-reagent-data.html）了解更多关于此试剂的信息。
RNAi可以使用两种不同方法导入——合成的siRNA双链或通过质粒或病毒载体表达的siRNA（shRNA，miRNAi）。siRNA正成为快速发展的治疗应用的首选方法。它们易于使用，易于设计，并且易于合成。siRNA可以被快速鉴定并且可以同时靶向多个基因。对于RNAi载体而言，其表达可能由于质粒在基因组的稳定整合而更稳定持久，而它们也可以靶向非分裂细胞例如干细胞、淋巴细胞和神经元。RNAi载体的不足是插入突变导致的致癌风险，以及长期基因沉默和/或细胞内大量siRNA的存在导致的不可预测的毒性（Grimm D. et al.: Nature 441: 537-541 (2006)）。
对于 Stealth siRNA我们推荐选用HPLC in vivo纯化级别。对于Ambion In Vivo siRNA，我们推荐in vivo ready纯化级别。
我们已经证明标记的Stealth和Silencer siRNA不会影响它们的基因敲低能力。一个替代的方法是可以将非标记siRNA的和标记的对照siRNA混合；该方法更多用于体内siRNA实验，并可以使临床研究得以推进而不受标记可能带来的效应的影响。
化学修饰的siRNA双链相比标准siRNA双链有很多优势，包括低脱靶效应，高稳定性，以及低毒性。因为这些原因，我们推荐使用化学修饰的siRNA用于体内实验。对于一些目的只是确定生物分布的探索实验，非修饰的siRNA和荧光对照会更有帮助并且稍微便宜一些。
•比非修饰siRNA稳定性>100倍——体内基因敲低效果更持久
•兼容于任何21-mer的序列——敲低能力不会损失
•无毒——实际上降低了siRNA序列特异的免疫反应
•高特异性——脱靶效应最高能降低90%
•提供预设计和经验证两种Ambion In Vivo siRNA
请参考下列用于体内研究的不同类型的siRNA：
AmbionIn Vivo预设计siRNA使用Silencer Select算法设计并带有额外化学修饰以获得超强血清稳定性（在37°C下90%小鼠血清中的半衰期>5小时）从而适用于体内实验。 AmbionIn Vivo siRNA在体外（外周血单核细胞；PBMC）和体内（小鼠）均无毒和无免疫源性。在细胞实验中，AmbionIn Vivo siRNA表现出和Silencer Select siRNA相当的效果和特异性。
定制设计的Ambion In VivosiRNA针对您的特定目的基因而设计，使用Silencer Select算法并带有Ambion In Vivo化学修饰。
定制合成的Ambion In VivosiRNA根据您提供的序列合成，带有Ambion In Vivo化学修饰以提高血清稳定性。
我们的AmbionIn Vivo siRNA可以以下列几种纯化级别订购：
脱盐：脱盐并使用MALDI-TOF质谱分析
HPLC：HPLC纯化并使用MALDI-TOF和分析级HPLC分析
In Vivo Ready：HPLC纯化，使用MALDI-TOF和分析级HPLC分析，透析脱盐，过滤灭菌，并经内毒素检测。
使用我们的GeneAssist工具（https://www.thermofisher.com/order/custom-genomic-products/tools/sirna/）订购您的定制siRNA。
阳性对照siRNA例如Silencer Select GAPDH siRNA将证明转染的有效性并有助于优化转染条件。阴性对照siRNA对于设定基因表达的基线是必要的。
对于Silencer和Silencer Select siRNA我们均提供多个阴性和阳性对照。
Silencer siRNA阳性对照有：
- Silencer Firefly Luciferase (GL2 + GL3) siRNA（货号AM4629, 5nmol）
- Silencer GAPDH siRNA（人）（货号AM4605, 5nmol; AM4633, 40 nmol）
- Silencer GAPDH siRNA (人，小鼠，大鼠) (货号AM4624, 5 nmol; AM4631, 40 nmol; AM4632, 5 x 40 nmol)
- Silencer GFP (eGFP) siRNA （货号AM4626, 5 nmol）
- Silencer KIF11 (Eg5) siRNA (人，小鼠，大鼠) (货号AM4639, 5 nmol）
如果使用荧光标记的Silencer siRNA，我们有：
- Silencer Cy-3标记GAPDH siRNA (人，小鼠，大鼠) (货号AM4649, 5 nmol)
- Silencer FAM-标记GAPDH siRNA(人，小鼠，大鼠) (货号AM4650, 5 nmol)
如果使用体内级别的Silencer siRNA，可以选：
- Silencer GAPDH阳性对照siRNA，in vivo ready（货号4404025, 250 nmol）
Silencer siRNA阴性对照有：
- Silencer阴性对照1号 siRNA（货号AM4611, 5 nmol; AM4635, 40 nmol; AM4636, 5 x 40 nmol）
- Silencer阴性对照2号siRNA（货号AM4613, 5 nmol; AM4637, 30 nmol）
- Silencer阴性对照3号 siRNA（货号AM4615, 5 nmol）
- Silencer阴性对照4号 siRNA（货号AM4641, 5 nmol）
- Silencer阴性对照5号 siRNA（货号AM4642, 5 nmol）
如果使用荧光标记的Silencer siRNA，我们有：
- SilencerCy 3-标记阴性对照1号siRNA（货号AM4621, 5 nmol）
- Silencer FAM-标记阴性对照1号 siRNA（货号AM4620, 5 nmol）
如果使用体内级别的Silencer siRNA，可以选：
- Silencer 阴性对照 1号 siRNA（货号4404021）
Silencer Select siRNA阳性对照有：
- Silencer Select GAPDH阳性对照siRNA（货号4390849, 5 nmol; 4390850, 40 nmol; 4404024, 250 nmol）
- Silencer Select MALAT1阳性对照siRNA，靶向非编码siRNA (货号4455877, 5 nmol)
如果使用体内级别的Silencer Select siRNA，可以选：
- Silencer Select GAPDH in vivo（货号44404024）
Silencer Select siRNA阴性对照有：
- Silencer Select阴性对照1号 siRNA (货号4390843, 5 nmol; 4390844, 40 nmol)
- Silencer Select阴性对照2号 siRNA (货号4390846, 5 nmol; 4390847, 40 nmol)
如果使用体内级别的Silencer Select siRNA，可以选：
- Silencer Select阴性对照1号 siRNA, 体内级别 (货号4404020, 250nmol)
We offer the following in vivo siRNA controls:
Invitrogen In Vivo Negative Control #1 siRNA (Cat. No. 4457287, 5nmol; 4459405, 50 nmol; 4457289, 250 nmol)
Invitrogen In Vivo GAPDH Positive control siRNA (Cat. No. 4457288, 5 nmol; 4459407, 50 nmol, 4457291, 250 nmol)
Invitrogen In Vivo Factor VII Positive control siRNA (Cat. No. 4459408, 50 nmol; 4457292, 250 nmol)
Silencer Select Negative Control No. 1 siRNA, in vivo ready (Cat. No. 4404020, 250 nmol)
Silencer GAPDH Positive Control siRNA, in vivo ready (Cat. No. 4404025, 250 nmol)
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
Stealth siRNA is markedly more stable in serum than unmodified siRNA. This increased stability is particularly beneficial in vivo where the RNAi may be exposed to more proteases. In collaboration with Intradigm, Stealth RNAi has been demonstrated to have activity in vivo when injected intra-tumorally.
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
Pre-designed Stealth RNAi and Ambion In Vivo siRNA can be ordered directly from our pre-designed siRNA search interface (https://www.thermofisher.com/us/en/home/life-science/rnai/introduction-to-in-vivo-rnai.html?open=faq#search) to target human, mouse, or rat genes.
Custom siRNA designs for in vivo experiments can be ordered from two interfaces, depending on the product format being requested: Custom Ambion In Vivo siRNA is ordered from the GeneAssist Custom siRNA Builder (https://www.thermofisher.com/order/custom-genomic-products/tools/sirna/), while Stealth RNAi is ordered using the BLOCK-iT RNAi Express for in vivo synthetics (https://rnaidesigner.thermofisher.com/rnaiexpress/inVivoService.jsp).
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
It is recommended to reduce the chance of eliciting an immune response.
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
Please contact rnairesearcher@invitrogen.com regarding large scale in vivo siRNA or fill out the quote request form (www.thermofisher.com/us/en/home/global/forms/custom-sirna-mirna-quote-request.html).
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
Several different approaches have been used for siRNA delivery, including various local delivery techniques and systemic delivery. We offer a lipid reagent, Invivofectamine 3.0 Transfection Reagent for in vivo siRNA systemic delivery. Learn more about this reagent here (https://www.thermofisher.com/us/en/home/life-science/rnai/introduction-to-in-vivo-rnai/invivofectamine-reagent.html).
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
RNAi can be delivered using two different approaches-siRNA synthetic duplexes or siRNA expressed from plasmids or viral vectors (shRNA, miRNAi). siRNA are becoming the method of choice for the fast development of therapeutics. They are easy to use, easy to design, and easy to synthesize. siRNA can be rapidly identified and multiple genes can be targeted at the same time. With RNAi vectors, the expression will be steadier as a result of the possibility of stable integration of the plasmid into the genome, and they have the ability to target nondividing cells such as stem cells, lymphocytes and neurons. The drawbacks are the danger of oncogenic transformation from insertional mutagenesis, and unanticipated toxicity from long-term silencing of human genes and/or having high amounts of siRNA inside the cell (Grimm D. et al.: Nature 441: 537-541 (2006)).
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
For Stealth siRNA we recommend to select HPLC in vivo purity. For Invitrogen In Vivo siRNA, we recommend in vivo ready purity.
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
We have demonstrated that labeling Stealth and Silencer siRNA does not hamper their knockdown potency. An alternative approach is to mix unlabeled duplexes with labeled control duplexes; this method is more commonly used with in vivo siRNA, and allows progression to clinical research unhindered by questions about the possible effects of a label.
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
Chemically modified siRNA duplexes have a number of advantages over standard siRNA duplexes, including the minimization of off-target effects, enhanced stability, and reduced toxicity. For these reasons, chemically modified siRNA duplexes are recommended for in vivo siRNA experiments. For some pilot experiments, where the only aim is to determine biodistribution, unmodified siRNA and fluorescent controls are useful and slightly more cost-effective.
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
- 100x more stable than unmodified siRNA - longer knockdown in vivo
- Compatible with any 21-mer sequence - no loss of potency
- Non-toxic - actually reduces siRNA sequence specific immune responses
- Enhanced specificity - up to 90% reduction in off target effects
- Both Pre-designed and Validated Invitrogen In Vivo siRNAs offered
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
Please see the different siRNA we offer for in vivo studies:
Ambion In Vivo Pre-designed siRNAs are designed using the Silencer Select algorithm and incorporate additional chemical modifications for superior serum stability (half life >5 hours at 37 degrees C in 90% mouse serum) with in vivo applications. Ambion In Vivo siRNAs are non-toxic and non-immunogenic in vitro (peripheral blood nononuclear cells; PBMC) and in vivo (mouse). In cell-based assays, Ambion In Vivo siRNAs exhibit potency and specificity equivalent to Silencer Select siRNAs.
Ambion In Vivo Custom Designed siRNAs are designed to your specified target, using the Silencer Select algorithm and Ambion In Vivo chemical modifications.
Ambion In Vivo Custom Synthesized siRNAs are synthesized with your sequence, with Ambion In Vivo chemical modifications for serum stability.
Our Ambion In Vivo siRNA can be ordered with the following purities:
Desalted: Desalted and analyzed by MALDI-TOF mass spectrometry.
HPLC: HPLC purified and analyzed by MALDI-TOF and analytical HPLC
In Vivo Ready : HPLC purified, analyzed by MALDI-TOF and analytical HPLC, dialyzed to remove salts, sterile filtered, and endotoxin tested
Use our GeneAssist tool to order your custom siRNA (http://www.thermofisher.com/order/custom-genomic-products/tools/sirna/).
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
A positive control siRNA such as the Silencer Select GAPDH siRNA will demonstrate efficient transfection and help to optimize transfection conditions. A negative control siRNA is necessary to serve as a baseline for gene expression.
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
We offer several negative and positive controls for both Silencer and Silencer Select siRNA.
For Silencer siRNA positive control:
- Silencer Firefly Luciferase (GL2 + GL3) siRNA (Cat. No. AM4629, 5nmol)
- Silencer GAPDH siRNA (human) (Cat. No AM4605, 5nmol; AM4633, 40 nmol)
- Silencer GAPDH siRNA (human, mouse, rat) (Cat. No. AM4624, 5 nmol; AM4631, 40 nmol; AM4632, 5 x 40 nmol)
- Silencer GFP (eGFP) siRNA (Cat. No. AM4626, 5 nmol)
- Silencer KIF11 (Eg5) siRNA (human, mouse, rat) (Cat. No. AM4639, 5 nmol)
If working with Fluorescently labeled Silencer siRNA:
- Silencer Cy-3 labeled GAPDH siRNA (human, mouse, rat) (Cat. No. AM4649, 5 nmol)
- Silencer FAM-labeled GAPDH siRNA (human, mouse, rat) (Cat. NO. AM4650, 5 nmol)
If working with Silencer siRNA in vivo ready:
- Silencer GAPDH Positive Control siRNA, in vivo ready (Cat. No. 4404025, 250 nmol)
For Silencer siRNA negative controls:
- Silencer Negative Control No. 1 siRNA (Cat. Nos. AM4611, 5 nmol; AM4635, 40 nmol; AM4636, 5 x 40 nmol)
- Silencer Negative Control No. 2 siRNA (Cat. Nos. AM4613, 5 nmol; AM4637, 30 nmol)
- Silencer Negative Control No. 3 siRNA (Cat. No. AM4615, 5 nmol)
- Silencer Negative Control No. 4 siRNA (Cat. No. AM4641, 5 nmol)
- Silencer Negative Control No. 5 siRNA (Cat. No. AM4642, 5 nmol)
If working with Fluorescently labeled Silencer siRNA:
- Silencer Cy 3-labeled Negative Control No. 1 siRNA (Cat. No AM4621, 5 nmol)
- Silencer FAM-labeled Negative Control No. 1 siRNA (Cat. No. AM4620, 5 nmol)
If working with Silencer siRNA in vivo ready:
- Silencer in vivo ready (Cat. No. 4404021)
For Silencer Select siRNA Positive control:
- Silencer Select GAPDH Positive Control siRNA (Cat. No. 4390849, 5 nmol; 4390850, 40 nmol; 4404024, 250 nmol)
- Silencer Select MALAT1 Positive Control siRNA for non-coding siRNA (Cat. No. 4455877, 5 nmol)
For Silencer Select siRNA negative control:
- Silencer Select Negative Control No. 1 siRNA (Cat. No. 4390843, 5 nmol; 4390844, 40 nmol)
- Silencer Select Negative Control No. 2 siRNA (Cat. No. 4390846, 5 nmol; 4390847, 40 nmol)
If working with Silencer Select siRNA in vivo ready:
- Silencer Select Negative Control No. 1 siRNA, in vivo ready (Cat. No. 4404020, 250nmol)
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.