Easy transgenesis means easy reprogramming
Easily and efficiently deliver the miR-302/367 cluster for iPSC generation using the PB-Cuo-miR-302/367-IRES-GFP-EF1α-CymR-Puro Inducible Human iPSC Vector. This pre-built vector is ready-to-co-transfect with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1), and expresses miR-302a, miR-302b, miR-302c, miR-302d, and miR-306 with SBI’s cumate-inducible promoter. The vector also includes a co-expressed GFP reporter to simplify selection of transfectants, and the EF1α promoter co-expressing the cumate repressor (CymR) and puromycin resistance.
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)
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.
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
Robust, cumate-inducible expression of the miR-302/367 cluster
Figure 2. Robust, cumate-inducible expression of the miR-302/367 cluster. The PiggyBac miR-302/367 iPSC Vector (Cat.# PBQM-miR302) was co-transfected with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) into 293T cells. After three days growth to enable integration of the PiggyBac insert into the genome, induction was initiated by adding cumate to a final concentration of 30 µg/mL. Three days later, cells were harvested and expression levels of the miR-302/367 miRNAs measured using SBI’s qPCR-based QuantiMir Kit (Cat.# RA420A-1). miR-302/367 expression levels were normalized to the U6 control to determine the fold induction.