The piggyBac-Based Gene Delivery System Can Confer Successful Production of Cloned Porcine Blastocysts with Multigene Constructs

摘要: The introduction of multigene constructs into single cells is important for improving the performance domestic animals, as well understanding basic biological processes. In particular, allow engineering and integration multiple genes related to xenotransplantation porcine genome. piggyBac (PB) transposon system allows be stably integrated target genomes through a transfection event. However, our knowledge, no attempt introduce genome has been made using this system. study, we simultaneously introduced seven transposons embryonic fibroblast (PEF). PEFs were transfected with containing five drug resistance proteins two (red green) fluorescent proteins, together PB transposase expression vector, pTrans (experimental group). above (without pTrans) concomitantly (control Selection these in presence selection drugs resulted survival several clones derived from experimental group, but not control. PCR analysis demonstrated that approximately 90% (12/13 tested) surviving possessed all transposons. Splinkerette inserted TTAA sites PB. Somatic cell nuclear transfer (SCNT) PEF clone successful production cloned blastocysts expressing both red green fluorescence. These results indicate feasibility PB-mediated method simultaneous genome, which useful transgenic pigs transgenes.