Adeno‐associated virus (AAV)‐mediated transduction of male germ line stem cells results in transgene transmission after germ cell transplantation

Abstract

We explored whether exposure of mammalian germ line stem cells to adeno-associated virus (AAV), a gene therapy vector, would lead to stable transduction and transgene transmission. Mouse germ cells harvested from experimentally induced cryptorchid donor testes were exposed in vitro to AAV vectors carrying a GFP transgene and transplanted to germ cell-depleted syngeneic recipient testes, resulting in colonization of the recipient testes by transgenic donor cells. Mating of recipient males to wild-type females yielded 10% transgenic offspring. To broaden the approach to nonrodent species, AAV-transduced germ cells from goats were transplanted to recipient males in which endogenous germ cells had been depleted by fractionated testicular irradiation. Transgenic germ cells colonized recipient testes and produced transgenic sperm. When semen was used for in vitro fertilization (IVF), 10% of embryos were transgenic. Here, we report for the first time that AAV-mediated transduction of mammalian germ cells leads to transmission of the transgene through the male germ line. Equally important, this is also the first report of transgenesis via germ cell transplantation in a nonrodent species, a promising approach to generate transgenic large animal models for biomedical research.

Induced Pluripotent Stem Cell Clones Reprogrammed via Recombinant Adeno-Associated Virus-Mediated Transduction Contain Integrated Vector Sequences

“…This suggests that DNA breakage prior to or during iPSC induction might facilitate AAV integration. In support of this view, rAAV was shown to efficiently transduce male germ line stem cells, which undergo meiotic divisions and are accompanied by a frequent occurrence of DSBs, thus resulting in stable transgene transmission through generations (1,11). In addition, AAV genomes have been shown to integrate preferentially at actively transcribed genes in neonatal and adult mouse liver, and these integration events are frequently associated with chromosomal deletions (20).…”

Recent advances in the development of new transgenic animal technology

“…Transgenic germ cells colonized recipient testes and produced transgenic sperm. When semen was used for in vitro fertilization (IVF), 10 % of embryos were transgenic [36]. This study was the first attempt to produce large transgenic domestic animal embryos via SSCs transplantation and is encouraging for the production of large transgenic animals.…”

Can we grow sperm? A translational perspective on the current animal and human spermatogenesis models

“…[10][11][12][13] Furthermore, by modifying the SSC prior to transplantation, transgenic animals can be generated using this technique. [14][15][16][17] Clinically, there may be potential to apply the SSC transplantation model in humans, especially in the fertility preservation of treated pre-pubertal male cancer patients (as outlined in Figure 1 18 ). In postpubertal patients, mature sperm can be easily banked prior to cancer therapies.…”