The 4-in-1 nonviral PiggyBac™ transposon vectors from SBI enable the reprogramming of human and mouse cells. Featuring a 2A-mediated coexpression cassette of human/murine c-Myc, Klf4, Oct4 and Sox2 iPSC genes, reprogram somatic cells into induced pluripotent stem cells with the highly efficient PiggyBac™ transposon system.
Reprogramming of human cells with PiggyBac™
Human neonatal skin fibroblasts (1x10^5 cells) were cotransfected using electroporation with 2.5 ug PiggyBac 4-in-1 vector (PB630A-1) and 0.5 ug Super PiggyBac transposase (PB210A-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.
Reprogramming of mouse cells with PiggyBac™
Panel A Gene Expression analysis of 4-factor PiggyBac transposon vector was tested in 3T3 Cells transfected with either no plasmid (-) or 2ug PiggyBac 4-in-1 vector and 0.5ug PiggyBac transposase (+). Cells were harvested two-weeks post-transfection, RNA was isolated and reverse transcribed to cDNA. Gene expression was evaluated via PCR using primers specific for 4-factors and Gapdh loading control. Panel B 3T3 cells were visualised two weeks post-transfection of 2ug PiggyBac 4-in-1 transposon vector + 0.5ug PiggyBac transposase for GFP expression and -/+ stained for anti-Sox2 antibody (Texas Red). DAPI stain was used as a positive control to identify cell nuclei.
Material available for download
PiggyBac Transposon Vector System datasheet
PiggyBac Transposon Vector System user manual
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