Research Article

Application of the Sleeping Beauty system in Saanen goat fibroblast cells for establishing persistent transgene expression

Published: December 08, 2011
Genet. Mol. Res. 10 (4) : 3347-3355 DOI: https://doi.org/10.4238/2011.October.25.4
Cite this Article:
B.C. Jiang, H.A. Kaleri, H.X. Zhang, J. Chen, H.L. Liu (2011). Application of the Sleeping Beauty system in Saanen goat fibroblast cells for establishing persistent transgene expression. Genet. Mol. Res. 10(4): 3347-3355. https://doi.org/10.4238/2011.October.25.4
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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.

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.