Assessing Tn5 and Sleeping Beauty for transpositional transgenesis by cytoplasmic injection into bovine and ovine zygotes

摘要: Transgenic domestic animals represent an alternative to bioreactors for large-scale production of biopharmaceuticals and could also provide more accurate biomedical models than rodents. However, their generation remains inefficient. Recently, DNA transposons allowed improved transgenesis efficiencies in mice pigs. In this work, Tn5 Sleeping Beauty (SB) transposon systems were evaluated by simple cytoplasmic injection livestock zygotes. the case Tn5, transposome complex nucleic acid protein was injected. SB, supercoiled plasmids encoding a SB transposase co-injected. vitro produced bovine zygotes used establish conditions. The cultured blastocysts reporter gene expression genotyped. Subsequently, both injected seasonally available ovine zygotes, employing carrying recombinant human factor IX driven beta-lactoglobulin promoter. approach did not result transgenic lambs. contrast, resulted 2 lambs (29%) transgene. Both exhibited cellular mosaicism extraembryonic tissues (placenta or umbilical cord) three additional transgenic. These results show that transpositional components can be applied pharmaceutical interest.