EGFP

Production of human factor VIII-FL in 293T cells using the bicistronic MGMT(P140K)-retroviral vector

A. M. Fontes, Melo, F. U. F., Greene, L. J., Faça, V. M., Lin, Y., Gerson, S. L., and Covas, D. T., Production of human factor VIII-FL in 293T cells using the bicistronic MGMT(P140K)-retroviral vector, vol. 11, pp. 775-789, 2012.

Hemophilia A is the most common X-linked bleeding disorder; it is caused by deficiency of coagulation factor VIII (FVIII). Replacement therapy with rFVIII produced from human cell line is a major goal for treating hemophilia patients. We prepared a full-length recombinant FVIII (FVIII-FL), using the pMFG-P140K retroviral vector. The IRES DNA fragment was cloned upstream to the P140K gene, providing a 9.34-kb bicistronic vector. FVIII-FL cDNA was then cloned upstream to IRES, resulting in a 16.6-kb construct.

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

B. C. Jiang, Kaleri, H. A., Zhang, H. X., Chen, J., and Liu, H. L., Application of the Sleeping Beauty system in Saanen goat fibroblast cells for establishing persistent transgene expression, vol. 10, pp. 3347-3355, 2011.

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.

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