PB-EF1-MCS-IRES-RFP cDNA Cloning and Expression Vector

One transfection can integrate one or more genes that can be precisely removed

Figure 2. Efficient transgenesis with the Super PiggyBac Transposase and both single- and dual-promoter PiggyBac Vectors. (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.

Figure 3. Simultaneous integration of multiple PiggyBac Vectors is also highly efficient. METHODS: Three different PiggyBac transposon vectors (Cat.# PB513B-1, Cat.# PB533A-2, and Cat.# PB531A-2) were co-transfected with (left panels) or without (right panels) the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) into Human HT1080 cells. Puromycin and neomycin selection was applied for seven days. The cells that were co-transfected with the Super PiggyBac Transposase Expression Vector were puro and neo resistant, GFP-positive, and RFP-positive. Background GFP-positive cells that are puro resistant stem from random PB513B-1 integrations during the puromycin selection. The non-PiggyBac-mediated integration rate in those cells was extremely low and no RFP-positive cells were identified.