Transposon iPSC reprogramming system

The piggyBac Transposon System from System Biosciences (SBI) enables large insert capabilities to simultaneously deliver four reprogramming factors for iPSC generation. These 4-in-1 iPSC vectors enable the reprogramming of human and mouse cells. 

How the piggyBac transposon system works
The pre-built vector is ready-to-co-transfect with the Super PiggyBac Transposase Expression Vector, and delivers cMyc, Klf4, Oct4, and Sox2 co-expressed from a strong CAGs promoter for mouse and delivers Oct4, Sox2, Klf4, and Myc co-expressed from a moderate EF1α promoter for human. The vector also includes a GFP reporter to simplify selection of transfectants.

Benefits of using the piggyBac transposon system
• Create 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

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.

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.

Please note: 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:

• End-user license information fro academic customers
• End-user license information for commercial customers
   

Material available for download
PiggyBac Transposon Vector System datasheet
PiggyBac Transposon Vector System user manual

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