PB-Cuo-shMCS-IRES-GFP-EF1α-CymR-Puro Inducible shRNA Cloning and Expression Vector

Combining easy PiggyBac transgenesis with our robust and titratable cumate-inducible expression system, this PiggyBac Vector is designed for shRNA expression

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PB-Cuo-shMCS-IRES-GFP-EF1α-CymR-Puro Inducible shRNA Cloning and Expression Vector

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PB-Cuo-shMCS-IRES-GFP-EF1α-CymR-Puro Inducible shRNA Cloning and Expression Vector

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10 µg
PBQMSH812A-1
$ 921
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Overview

Robust, titratable shRNA expression delivered using the PiggyBac Transposon System

More than just easy, consistent transgenesis with PiggyBac, the PB-Cuo-shMCS-IRES-GFP-EF1α-CymR-Puro Inducible shRNA Cloning and Expression Vector (Cat.# PBQMSH812A-1) adds in the robust and titratable gene expression control of SBI’s cumate-inducible expression system. Clone your shRNA-of-interest into the shMCS for cumate-inducible expression, which you can quantitatively monitor with the co-expressed GFP. This vector also co-expresses the cumate repressor, CymR, and puromycin resistance from the EF1α promoter.

PB-Cuo-shMCS-IRES-GFP-EF1α-CymR-Puro Inducible shRNA Cloning and Expression Vector

With the PiggyBac Transposon System, you can:

  • Make transgenic cell lines with a single transfection
  • Integrate multiple PiggyBac Vectors in a single transfection
  • Insert an expression cassette into human, mouse, and rat cells
  • Deliver virtually any-sized DNA insert, from 10 – 100 kb
  • Choose from PiggyBac Vectors that express your gene-of-interest from constitutive or inducible promoters and include a variety of markers
  • Determine the number of integration events with the PiggyBac qPCR Copy Number Kit (# PBC100A-1)

Customer Agreements
Academic customers can purchase PiggyBac Transposon System components for internal research purposes for indefinite use, whereas commercial customers must sign a customer agreement for a six-month, limited-use license to test the technology.
For end user license information, see the following:

* SBI is fully licensed to distribute PiggyBac vectors as a partnership with Transposagen Biopharmaceuticals, Inc.

How It Works

The PiggyBac Transposon System’s Cut-and-Paste Mechanism

The efficient PiggyBac Transposon System uses a cut-and-paste mechanism to transfer DNA from the PiggyBac Vector into the genome. If only temporary genomic integration is desired, the Excision-only PiggyBac Transposase can be transiently expressed for footprint-free removal of the insert, resulting in reconstitution of the original genome sequence.

The PiggyBac Transposon System’s cut-and-paste mechanism

Figure 1. The PiggyBac Transposon System’s cut-and-paste mechanism.

  • The Super PiggyBac Transposase binds to specific inverted terminal repeats (ITRs) in the PiggyBac Cloning and Expression Vector and excises the ITRs and intervening DNA.
  • The Super PiggyBac Transposase inserts the ITR-Expression Cassette-ITR segment into the genome at TTAA sites.
  • The Excision-only Super PiggyBac Transposase can be used to remove the ITR-Expression Cassette-ITR segment from the genome, for footprint-free removal

Tightly-controlled, inducible gene expression

Get robust, titratable gene expression with low background using SBI’s cumate-inducible vectors. These vectors take advantage of CymR, a repressor that binds to cumate operator sequences (CuO) with high affinity in the absence of cumate, a non-toxic small molecule. Providing much lower background expression than similar systems, SBI’s cumate-inducible vectors can provide up to 32-fold induction of gene expression.How the cumate operator switch works

  • 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
  • Powerful—suitable for in vivo applications

Supporting Data

Tight expression control with low background

In the absence of cumate, the cumate-inducible PiggyBac Vector shows undetectable levels of expression

Figure 2. In the absence of cumate, the cumate-inducible PiggyBac Vector shows undetectable levels of expression.

The PiggyBac cumate switch is titratable and can be turned off

Figure 3. The PiggyBac cumate switch is titratable and can be turned off.


Citations

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  • Cammareri, P, et al. (2017) TGFβ pathway limits dedifferentiation following WNT and MAPK pathway activation to suppress intestinal tumourigenesis. Cell Death Differ.. 2017 Jun 16;. PM ID: 28622298
  • Kimura, T, et al. (2017) Hsp90 inhibitor geldanamycin attenuates the cytotoxicity of sunitinib in cardiomyocytes via inhibition of the autophagy pathway. Toxicol. Appl. Pharmacol.. 2017 Jun 15; 329:282-292. PM ID: 28624441
  • 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
  • Liu, H, et al. (2017) Stochastic Protein Labeling Enables Long-Term Single Molecule Observation in vivo. bioRxiv. 2017 Jun 5;. Link: bioRxiv
  • 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
  • Lin, W & Li, Z. (2017) Blueberries inhibit cyclooxygenase-1 and cyclooxygenase-2 activity in human epithelial ovarian cancer. Oncol Lett. 2017 Jun 1; 13(6):4897-4904. PM ID: 28599493
  • Yamazaki, T, et al. (2017) Targeted DNA methylation in pericentromeres with genome editing-based artificial DNA methyltransferase. PLoS ONE. 2017 May 25; 12(5):e0177764. PM ID: 28542388
  • 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
  • Cruz-Molina, S, et al. (2017) PRC2 Facilitates the Regulatory Topology Required for Poised Enhancer Function during Pluripotent Stem Cell Differentiation. Cell Stem Cell. 2017 May 4; 20(5):689-705.e9. PM ID: 28285903
  • Wang, L, et al. (2017) Derivation and characterization of primordial germ cells from Guangxi yellow-feather chickens.. Poult. Sci.. 2017 May 1; 96(5):1419-1425. PM ID: 28158811
  • Uehara, T, et al. (2017) Selective degradation of splicing factor CAPER [alpha] by anticancer sulfonamides. Nature Chemical Biology. 2017 Apr 27; 13:675–680. Link: Nature Chemical Biology
  • 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
  • Wen, Y, et al. (2017) A stable but reversible integrated surrogate reporter for assaying CRISPR/Cas9-stimulated homology-directed repair. J. Biol. Chem.. 2017 Apr 14; 292(15):6148-6162. PM ID: 28228480
  • Mitra, A, et al. (2017) IL6-mediated inflammatory loop reprograms normal to epithelial-mesenchymal transition(+) metastatic cancer stem cells in preneoplastic liver of transforming growth factor beta-deficient β2-spectrin(+/-) mice. Hepatology. 2017 Apr 1; 65(4):1222-1236. PM ID: 27863449
  • Zamboni, CG, et al. (2017) Polymeric nanoparticles as cancer-specific DNA delivery vectors to human hepatocellular carcinoma. J Control Release. 2017 Mar 27;. PM ID: 28351668
  • Katayama, H, et al. (2017) Generation of non-viral, transgene-free hepatocyte like cells with piggyBac transposon. Sci Rep. 2017 Mar 15; 7:44498. PM ID: 28295042
  • Xu, X, et al. (2017) Reversal of Phenotypic Abnormalities by CRISPR/Cas9-Mediated Gene Correction in Huntington Disease Patient-Derived Induced Pluripotent Stem Cells.. Stem Cell Reports. 2017 Mar 14; 8(3):619-633. PM ID: 28238795