title: 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.
authors.affiliation: Center for Animal Transgenesis and Germ Cell Research, University of Pennsylvania, 382 W. State Road, Kennett Square, PA 19348, USA.
authors.forename: Ali
authors.initials: A
authors.lastname: Honaramooz
authors.forename: Susan
authors.initials: S
authors.lastname: Megee
authors.forename: Wenxian
authors.initials: W
authors.lastname: Zeng
authors.forename: Margret M
authors.initials: MM
authors.lastname: Destrempes
authors.forename: Susan A
authors.initials: SA
authors.lastname: Overton
authors.forename: Jinping
authors.initials: J
authors.lastname: Luo
authors.forename: Hannah
authors.initials: H
authors.lastname: Galantino-Homer
authors.forename: Mark
authors.initials: M
authors.lastname: Modelski
authors.forename: Fangping
authors.initials: F
authors.lastname: Chen
authors.forename: Stephen
authors.initials: S
authors.lastname: Blash
authors.forename: David T
authors.initials: DT
authors.lastname: Melican
authors.forename: William G
authors.initials: WG
authors.lastname: Gavin
authors.forename: Sandra
authors.initials: S
authors.lastname: Ayres
authors.forename: Fang
authors.initials: F
authors.lastname: Yang
authors.forename: P Jeremy
authors.initials: PJ
authors.lastname: Wang
authors.forename: Yann
authors.initials: Y
authors.lastname: Echelard
authors.forename: Ina
authors.initials: I
authors.lastname: Dobrinski
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D005854:Germ Cells / Q000378:metabolism* / Q000637:transplantation*; D006041:Goats; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D023421:Models, Animal; D012671:Seminiferous Tubules / Q000378:metabolism; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000378:metabolism*; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes / Q000235:genetics*