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

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.

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 four-month, limited-use license to evaluate the technology.
For end user license information, see the following:

• Academic Customers—End-User License Information
• Commercial Customers—End-User License Information

* SBI is fully licensed to distribute PiggyBac vectors as a partnership with Hera BioLabs, 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.

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.

• 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

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

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

Resources

Citations

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