Application of the Sleeping Beauty system in Saanen goat fibroblast cells for establishing persistent transgene expression |
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Authors: | Jiang B C Kaleri H A Zhang H X Chen J Liu H L |
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Institution: | College of Animal Sciences and Technology, Nanjing Agricultural University, Nanjing, China. |
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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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