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-EF1α-Oct4-Sox2-Klf4-Myc-IRES-GFP Human 4-in-1 iPSC PiggyBac Vector—to generate human iPSCs. This pre-built vector is ready-to-co-transfect with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1), and delivers human Oct4, Sox2, Klf4, and Myc co-expressed from a moderate EF1α promoter. The vector also includes a 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 human cells with the PiggyBac Transposon System
Figure 2. Easily reprogram human cells with the PiggyBac Transposon System. Human neonatal skin fibroblasts were co-transfected with PB-EF1α-Oct4-Sox2-Klf4-Myc-IRES-GFP Human 4-in-1 iPSC PiggyBac Vector (Cat.# PB630A-1) and the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1). iPS cells were derived using morphological selection criteria. When cultured under standard human ES cell culture conditions, the morphology of the custom human iPS cells was identical to that of human ES cells. Additionally, the cells express the pluripotency markers Oct4, Nanog, SSEA3, TRA-1-60, and demonstrate strong endogenous alkaline phosphatase staining.