Easy transgenesis means easy reprogramming
With the large insert capabilities of the PiggyBac Transposon System and the system’s easy, consistent transgenesis, you can use a single vector—the PB-SV40-Neo-EF1α-GFP-CAGs-cMyc-Klf4-Oct4-Sox2 Mouse 4-in-1 iPSC Vector—to generate mouse iPSCs. This pre-built vector is ready-to-co-transfect with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1), and delivers mouse cMyc, Klf4, Oct4, and Sox2 co-expressed from a strong CAGs promoter. The vector also includes a neomycin resistance marker and GFP reporter to simplify selection of transfectants.
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
Easily reprogram mouse cells with the PiggyBac Transposon System
Figure 2. Easily reprogram mouse cells with the PiggyBac Transposon System. (A) Gene expression of the PiggyBac Mouse 4-in-1 iPSC Vector was analyzed in 3T3 Cells transfected with either (- lane) the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) alone or (+ lane) the Super PiggyBac Transposase Expression Vector and the PiggyBac Mouse 4-in-1 iPSC Vector. Cells were harvested at two weeks post-transfection, and RNA was isolated and reverse transcribed (RT) into cDNA. Gene expression was evaluated via PCR using primers specific for the four factors and a Gapdh loading control. Only the cells co-transfected with the PiggyBac Mouse 4-in-1 iPSC Vector showed expression of the transcripts. (B). Cells from (A) were also imaged for GFP expression and stained with anti-Sox2 antibody (Texas Red, antibody in the + panel only). DAPI stain was used as a positive control to identify cell nuclei.