Transgenesis in one transfection: PiggyBac

The PiggyBac™ Transposon system from System Biosciences (SBI) is an easy system that provides consistent transgenesis with no limits on cargo size.

The system consists of a PiggyBac Vector and the Super PiggyBac Transposase which recognises transposon-specific inverted terminal repeats (ITRs) and efficiently integrates the ITRs and intervening DNA into the genome at TTAA sites. The Super PiggyBac Transposase is delivered to the cell via the Super PiggyBac Transposase Expression Vector, which is co-transfected with one or more PiggyBac Vectors.

Benefits of using the PiggyBac transposon vector system
• 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

How it works

1) The Super PiggyBac Transposase binds to the inverted terminal repeats (ITRs) in the PiggyBac Cloning and Expression Vector and excised the ITRs and intervening DNA. 2) The Super PiggyBac Transposase inserts the ITR-Expression Cassete-ITR segment into the genome at TTAA sites. 3) (Optional) The Excision-only Super PiggyBac Transposase can be used to remove the ITR-Expression Cassete-ITR segment from the genome, for footprint-free removal.

Efficient transgenesis with the Super PiggyBac Transposase and both single- and dual-promoter PiggyBac Vector

(Top four panels) Co-transfection with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) and a Dual Promoter PiggyBac Cloning and Expression Vector (Cat.# PB513B-1) into HeLa cells demonstrates the efficient integration delivered by SBI’s PiggyBac Transposon System. After ten days of puromycin selection, only the cells co-transfected with the Super PiggyBac Transposase (+PB, right two panels) show robust growth and GFP fluorescence. (Bottom four panels) Co-transfection with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) and a Single Promoter PiggyBac Cloning and Expression Vector (Cat.# PB531A-2) into HEK293 cells further demonstrates the efficient integration delivered by SBI’s PiggyBac Transposon System. After seven days of growth, the majority of cells that received the Super PiggyBac Transposase Expression Vector (+PB, right two panels) were RFP positive.

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. Please contact our SBI specialists for further details.

Material available for download
PiggyBac Transposon Vector System product sheet
PiggyBac Transposon Vector System user manual

PiggyBac transposon system citations