Long-term health and germline transmission in transgenic cattle following transposon-mediated gene transfer

Abstract

BackgroundTransposon-mediated, non-viral gene delivery is a powerful tool for generating stable cell lines and transgenic animals. However, as multi-copy insertion is the preferred integration pattern, there is the potential for uncontrolled changes in endogenous gene expression and detrimental effects in cells or animals. Our group has previously reported on the generation of several transgenic cattle by using microinjection of the Sleeping Beauty (SB) and PiggyBac (PB) transposons and seeks to explore the long-term effects of this technology on cattle.ResultsTransgenic cattle, one female (SNU-SB-1) and one male (SNU-PB-1), reached over 36 months of age with no significant health issues and normal blood parameters. The detection of transgene integration and fluorescent signal in oocytes and sperm suggested the capacity for germline transmission in both of the founder animals. After natural breeding, the founder transgenic cow delivered a male calf and secreted milk containing fluorescent transgenic proteins. The calf expressed green fluorescent protein in primary cells from ear skin, with no significant change in overall genomic stability and blood parameters. Three sites of transgene integration were identified by next-generation sequencing of the calf’s genome.ConclusionsOverall, these data demonstrate that transposon-mediated transgenesis can be applied to cattle without being detrimental to their long-term genomic stability or general health. We further suggest that this technology may be usefully applied in other fields, such as the generation of transgenic animal models.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-4760-4) contains supplementary material, which is available to authorized users.

Application of Transposon Systems in The Transgenesis of Bovine Somatic And Germ Cells

“…As such, various characteristics are divided according to the type of transposon. Previous in vivo experiments in our laboratory have produced transgenic cattle using PB and SB and have shown germ line transmission for more than 6 years without health problems (23). The embryo results of this experiment showed that not only PB and SB, but also Tol2, are highly e cient and Tol2 can be used as an alternative to PB and SB.…”

Complete blood count and selected serum parameters of Venus transgenic rabbits

“…Moreover, all selected serum biochemistry parameters were also within physiological ranges, thus, rabbits had no any severe diseases ( Table 2 ). Normal serum biochemistry and hematology parameters were also observed in CAG-EGFP transgenic cattle (Yum et al . 2018), EGFP transgenic piglets (Carter et al .…”

Application of transposon systems in the transgenesis of bovine somatic and germ cells

“…Therefore, various characteristics are divided according to the type of transposon. Previous in vivo experiments in our laboratory have produced transgenic cattle using PB and SB, which have shown germline transmission for more than 6 years without health problems [ 25 ]. The embryo results of this experiment demonstrate that PB, SB, and Tol2 are highly efficient and that Tol2 can be used as an alternative to PB and SB.…”