Overview
Get tight, titratable gene expression
With SBI’s SparQ™ Cumate Switch System, you can get inducible gene expression in mammalian cells with a range of cloning and expression lentivectors. The pCDH-CuO-MCS-IRES-GFP SparQ Cloning and Expression Lentivector (Cat.# QM530A-2) co-expresses both your gene- or miRNA-of-interest and the RFP marker from the inducible cumate switch promoter, enabling measurement of induction by RFP fluorescence. Co-expression is mediated by an IRES element.
With SBI’s SparQ™ Cumate Switch System, you can get inducible gene expression in mammalian cells through the binding of cumate, a non-toxic small molecule, to CymR. CymR can be delivered to cells on its own lentivector (Cat.#s QM200PA/VA-2 and QM300PA/VA-1), or on the same lentivector as your gene-of-interest (Cat.#s QM800A-1 and QM812B-1). Expression levels of your gene-of-interest are tightly controlled and increase with increasing cumate concentration until maximum induction is reached—see as much as a 32-fold increase in gene expression. Even better, induction is reversible, so you can turn expression levels on and off. Delivering negligible background expression in the absence of cumate, the SparQ System is an excellent choice for achieving controlled levels of gene expression.
- Robust—increase expression up to 32-fold
- Adjustable—tune expression levels by titrating the amount of cumate
- Reversible—turn expression on, then off, then on again
- Versatile—choose from all-in-one formats that co-express CymR and your gene-of-interest, or two-vector systems where CymR is expressed from a different plasmid
- Powerful—suitable for in vivo applications
How It Works
Tightly-controlled, inducible gene expression
SBI’s SparQ Cumate Switch System delivers robust, titratable gene expression with low background through three components:
- Cumate, a non-toxic, small-molecule inducer
- CymR, a repressor that binds to cumate operator sequences in the absence of cumate
- SparQ Lentivector that contains an MCS to clone-in your gene-of-interest, the cumate inducible promoter with cumate operator sequences (CuO) upstream of the MCS, and one or more markers
CymR has a high binding affinity for cumate and, as more cumate is added, fewer CymR molecules bind to the CuO sequences in the promoter resulting in increased expression. Exhibiting much lower background expression than similar systems, SBI’s cumate-inducible vectors can provide up to 32-fold induction of gene expression.
Supporting Data
Tight expression control with low background with the SparQ Cumate Switch System
Figure 1. Get lower background and higher induction with the SparQ Cumate Switch System than other inducible systems.
Figure 2. Gene expression with the SparQ Cumate SwitchSystem can be turned on and off, then on again.
Figure 3. Gene expression with the SparQ Cumate Switch System is titratable, with increasing amounts of cumate inducing a linear increase in gene expression.
Figure 4. With the SparQ System, gene expression can also be titrated by increasing the amount of transduced SparQ lentivirus, even up to 30 MOI.
Resources
Citations
- Shinmura, K, et al. (2017) WDR62 overexpression is associated with a poor prognosis in patients with lung adenocarcinoma. Mol. Carcinog..2017 Aug 1; 56(8):1984-1991. PM ID:28277612
- Maegawa, KI, et al. (2017) The Highly Dynamic Nature of ERdj5 Is Key to Efficient Elimination of Aberrant Protein Oligomers through ER-Associated Degradation. Structure.2017 Jun 6; 25(6):846-857.e4. PM ID:28479060
- Park, TS, Kim, SW & Lee, JH. (2017) Efficient transgene expression system using a cumate-inducible promoter and Cre-loxP recombination in avian cells. Asian-australas. J. Anim. Sci..2017 Jun 1; 30(6):886-892. PM ID:27764912
- Herrington, KA, et al. (2017) Spatial analysis of Cdc42 activity reveals a role for plasma membrane-associated Cdc42 in centrosome regulation. Mol. Biol. Cell.2017 May 24;. PM ID:28539409
- Qi, Z, et al. (2017) An optimized, broadly applicable piggyBac transposon induction system. Nucleic Acids Res..2017 Apr 20; 45(7):e55. PM ID:28082389
- Jain, A, et al. (2017) Abl kinase regulation by BRAF/ERK and cooperation with Akt in melanoma. Oncogene.2017 Apr 3;. PM ID:28368422
- O"Hara, SP, et al. (2017) ETS Proto-oncogene 1 Transcriptionally Up-regulates the Cholangiocyte Senescence-associated Protein Cyclin-dependent Kinase Inhibitor 2A. J. Biol. Chem..2017 Mar 24; 292(12):4833-4846. PM ID:28184004
- Ikushima, S & Boeke, JD. (2017) New Orthogonal Transcriptional Switches Derived from Tet Repressor Homologues for Saccharomyces cerevisiae Regulated by 2,4-Diacetylphloroglucinol and Other Ligands. ACS Synth Biol.2017 Mar 17; 6(3):497-506. PM ID:28005347
- Shinmura, K, et al. (2017) Reduced expression of the DNA glycosylase gene MUTYH is associated with an increased number of somatic mutations via a reduction in the DNA repair capacity in prostate adenocarcinoma. Mol. Carcinog..2017 Feb 1; 56(2):781-788. PM ID:27253753
- Liu, L, Huang, W & Huang, JD. (2017) Synthetic circuits that process multiple light and chemical signal inputs. BMC Syst Biol.2017 Jan 19; 11(1):5. PM ID:28103878
- Ando, T, et al. (2017) Ameloblastin induces tumor suppressive phenotype and enhances chemosensitivity to Dox via Src-Stat3 inactivation in osteosarcoma. Sci Rep.2017 Jan 5; 7:40187. PM ID:28054649
- Barta, T, Peskova, L & Hampl, A. (2016) miRNAsong: a web-based tool for generation and testing of miRNA sponge constructs in silico. Sci Rep.2016 Nov 18; 6:36625. PM ID:27857164
- Choi, JS, et al. (2016) cIAPs promote the proteasomal degradation of mutant SOD1 linked to familial amyotrophic lateral sclerosis. Biochem. Biophys. Res. Commun..2016 Nov 18; 480(3):422-428. PM ID:27773815
- Ushioda, R, et al. (2016) Redox-assisted regulation of Ca2+ homeostasis in the endoplasmic reticulum by disulfide reductase ERdj5. Proc. Natl. Acad. Sci. U.S.A..2016 Oct 11; 113(41):E6055-E6063. PM ID:27694578
- Welti, J, et al. (2016) Analytical Validation and Clinical Qualification of a New Immunohistochemical Assay for Androgen Receptor Splice Variant-7 Protein Expression in Metastatic Castration-resistant Prostate Cancer. Eur. Urol..2016 Oct 1; 70(4):599-608. PM ID:27117751
- Narumi, S, et al. (2016) SAMD9 mutations cause a novel multisystem disorder, MIRAGE syndrome, and are associated with loss of chromosome 7. Nat. Genet..2016 Jul 1; 48(7):792-7. PM ID:27182967
- Park, BO, et al. (2016) Selective novel inverse agonists for human GPR43 augment GLP-1 secretion. Eur. J. Pharmacol..2016 Jan 15; 771:1-9. PM ID:26683635
- Herrington, KA. (2016) New technologies for a better understanding of the Golgi: FLIM-FRET and click chemistry. Thesis.;. Link:Thesis
- Shinmura, K, et al. (2015) NEIL1 p.Gln282Stop variant is predominantly localized in the cytoplasm and exhibits reduced activity in suppressing mutations. Gene.2015 Oct 15; 571(1):33-42. PM ID:26095805
- Cheng, YS, et al. (2015) PPP2R5C Couples Hepatic Glucose and Lipid Homeostasis. PLoS Genet..2015 Oct 1; 11(10):e1005561. PM ID:26440364
SystemBiosciences，简称SBI，美国加利福尼亚湾区新成立的生技公司，致力于独特，创新生物技术之开发，以研发利于基因及蛋白质功能鉴定，研究之崭新方法和工具为宗旨。
美国SBI代理SystemBiosciences,简称SBI，美国加州湾区新成立的生技公司，致力于独特、创新生物技术之开发，以研发利于基因及蛋白质功能鉴定、研究之崭新方法和工具为宗旨。现阶段研发重心为RNA干扰（RNAi）研究之相关工具。
现阶段研发重心为RNA干扰（RNAi）的研究之相关工具。系统Biosciences公司（SBI）致力于开发独特，革新的技术，为客户研究蛋白组学和基因组学功能提供研究工具.SBI是专业的慢病毒产品公司，提供基于慢病毒的所有相关产品，质粒，试剂盒及相关配套试剂和慢病毒扩展产品，如IPS细胞多功能性诱导试剂盒和RNAi筛选文库。System Biosciences继续创造新的独特产品以及改善我们完善的产品线。我们对提供领先技术的承诺与我们所有研究试剂和研究项目服务的高质量制造和质量控制相匹配。 2009年要寻找的东西：以下是即将推出的新产品。新型miRZips™：基于慢病毒载体的新型技术可永久敲低MicroRNA。双标记shRNA表达载体：SBI将发布新的功能强大的shRNA表达慢病毒载体pGreenPuro™，以便为稳定的RNAi实验选择稳定转导的细胞的GFP和Puro标记。甾醇反应pGreenFire™慢病毒报告子：基于慢病毒载体的转录报告子，用于监测与固醇感应转录因子相关的转录网络活性-心血管疾病途径的关键。诱导型表达慢载体：新的构建体将允许CDNA，shRNA和microRNA的“按需”表达。2007–2008年的新产品新的miR-SNaRE：MicroRNA小型非编码RNA富集系统，该系统使用带有表位标签的microRNA加工因子来提取蛋白质及其相关RNA。识别驱动RISC复合体的信使RNA。新的GeneNet™聚焦的shRNA库：这些聚焦的shRNA库编码一组针对所有与人类激酶，磷酸酶或细胞凋亡相关基因有关的特定功能或类别基因设计的shRNA集合。这些文库可进行有针对性的高通量RNAi筛选。新的miRNomeMicroRNA分析仪：qPCR阵列在预先格式化的板中包括microRNA分析，可用于人类的完全互补或小鼠单个microRNA的完全互补，每块板上带有三个内源参考RNA对照。所有基于SangermiRBasemicroRNA数据库的microRNA分析均已注册。新的Lenti-miR™MicroRNA前体克隆：在基于HIV的慢病毒载体中可获得更多的microRNA前体集合。超过580个单独的microRNA前体克隆可用阵列形式或合并的慢病毒形式用于HT筛选。新的基于慢病毒的干细胞报道者：SBI越来越多的慢病毒载体已被开发为将基因构建体在体内外几乎传递给任何细胞类型的最有效方法。SBI已将我们的慢病毒构建体系列扩展为干细胞报道分子。使用连接到GFP报告基因的细胞和途径特异性启动子，轻松创建转基因细胞系以监测细胞分化。QuantiMir™RT试剂盒：这项流行的新技术可通过一次cDNA合成同时进行实时qPCR定量分析数百种microRNA。设计您自己的microRNA测定法以进行创新性实验。癌症microRNAqPCR分析小组（OncoMir系列）：预格式化的microRNA分析小组，用于评估95种已知与癌症有关的microRNA。干细胞microRNA分析小组：介绍了95种参与干细胞分化的microRNA，可同时监测干细胞的自我维持，造血途径和神经分化。SBI完善的产品线FullSpectrum™完整信使RNA扩增试剂盒：利用针对mRNA最常见基序的mRNA特异性引物提供完整的mRNA转录物（5"和3"末端）的无偏见，完整代表。GeneNet™siRNA库：全基因组，即用型，预包装的慢病毒库为筛选与生物学反应相关的基因功能提供了令人兴奋的机会。干扰素反应检测试剂盒：区分真正的RNA干扰和压力相关的细胞反应。PathNet™转录报告基因慢病毒载体：一种独特的方法，可创建各种稳定的报告基因细胞系，用于信号通路的研究。我们感谢您过去的支持，并期待为您提供最佳的新试剂，技术和服务，以加速您成功的研究目标。
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本是小菜鸟，目前研究某个比较新的蛋白对内质网应激的影响，需要抑制这个蛋白的表达看下游信号的表达情况，但是无论如何夜找不到抑制剂，做RNA干扰的话经费就超出预算了！急求各位大神，帮忙出个解决方案吧！非常感谢！
将EntransterTM-invivo与AmbioninvivosiRNA(作用于凝血因子VII)和阴性siRNA通过尾静脉注射成年小鼠，2天后，取动物肝脏检测。在mRNA水平和蛋白水平观察干扰效果。见上图，图中最左侧组为对照组注射阴性siRNA为3mg/kg，后边3组为每kg动物注射阳性siRNA量分别为1mg/kg,2mg/kg和3mg/kg情况。
根据推荐用量注射EntransterTM-invivo和siRNA（作用于LaminA/C）和阴性siRNA到成年小鼠。注射后2天收集相应的组织，分离RNA，用qRT-PCR分析LaminA/C基因的表达水平。图4为尾静脉结果，图5为各器官分别局部注射结果。
英格恩生物体内转染试剂，3天可完成动物体内转染实验，让动物干扰，基因敲除实验变得简单、快速有效！
简单来说，Off-target效应就是指干扰shRNA序列进入了microRNA途径，通过microRNA途径，其可以不受完全互补的限制而调控大量靶基因的表达。原本需要19~23nt的RNA序列完全互补才能发生干扰作用，而如果进入microRNA途径，只需要11~15nt互补就可以产生干扰效果，这使得siRNA可能与非靶基因结合而导致非靶基因沉默，造成脱靶。 如果脱靶干扰的部分基因，正好与目的基因位于同一信号通路中，或者与目的基因的生物学功能相似，那么，如果因脱靶而干扰了其它基因，亦会造成和目的基因受到干扰后相同的细胞表型改变。而实际上，可能选择的这条shRNA序列并没有对目的基因造成有效干扰，或者虽然干扰了目的基因，但是并不会引起预期的细胞表型改变。
基于以上原因，自从Off-target效应被发现并被研究者们广泛认同后，越来越多的杂志要求研究者们在投稿时需要有相应的对照来说明Off-target效应。即您需要有相关对照或者实验来说明，您所获得的实验结果，不是由于Off-target效应而产生的。向左转|向右转
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