The authors present a non-viral transfection method for reprogramming somatic cells to pluripotency, creating induced pluripotent stem (iPS) cells. They developed a single multiprotein expression vector containing the coding sequences of c-Myc, Klf4, Oct4, and Sox2 linked with 2A peptides, which can be used to reprogram both mouse and human fibroblasts. The transgene can be removed once reprogramming is achieved, minimizing genome modification in iPS cells. The iPS cells produced with this method show robust expression of pluripotency markers and can be functionally confirmed by in vitro differentiation assays and the formation of adult chimeric mice. When combined with a piggyBac transposon, the method successfully established reprogrammed human cell lines from embryonic fibroblasts. This system enables the complete elimination of exogenous reprogramming factors, providing iPS cells suitable for regenerative medicine, drug screening, and disease modeling.The authors present a non-viral transfection method for reprogramming somatic cells to pluripotency, creating induced pluripotent stem (iPS) cells. They developed a single multiprotein expression vector containing the coding sequences of c-Myc, Klf4, Oct4, and Sox2 linked with 2A peptides, which can be used to reprogram both mouse and human fibroblasts. The transgene can be removed once reprogramming is achieved, minimizing genome modification in iPS cells. The iPS cells produced with this method show robust expression of pluripotency markers and can be functionally confirmed by in vitro differentiation assays and the formation of adult chimeric mice. When combined with a piggyBac transposon, the method successfully established reprogrammed human cell lines from embryonic fibroblasts. This system enables the complete elimination of exogenous reprogramming factors, providing iPS cells suitable for regenerative medicine, drug screening, and disease modeling.