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Updated: May 28, 2026
Electroporation-Based Genetic Modification of Primary Human Pigment Epithelial Cells Using the Sleeping Beauty Transposon System
Published on: February 4, 2021
Application of the Sleeping Beauty system in Saanen goat fibroblast cells for establishing persistent transgene
B C Jiang1, H A Kaleri, H X Zhang
1College of Animal Sciences and Technology, Nanjing Agricultural University, Nanjing, China.
Abstract:
The Sleeping Beauty (SB) transposon system is a promising new method for establishing persistent transgene expression in vivo. We applied the SB system for enhancing transgenesis in Saanen dairy goat fibroblast cells. We constructed a pKT2/CMV-EGFP-IRES-PURO vector and investigated the influence of transposon and transposase vector ratios on transfection efficiency in the Saanen goat fibroblast cells. To enhance the SB system performance, we developed a new transfection technique (double-transfection method) for the SB system. The cultured cells were transfected with transposase and transposon vectors successively, with a 42-h interval. Consequently, the transposase and DNA donor (transposon vector) can interact, both at the highest level. Compared with the traditional transfection method, this new double-transfection method approximately doubled integration efficiency.
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