biostudies-literatureCiccarelli MUtah State UniversityWashington State UniversityUSDA | National Institute of Food and AgricultureBiotechnology and Biological Sciences Research Council24195-24204https://www.ebi.ac.uk/biostudies/studies/S-EPMC7533891biostudies-literatureUnknown117(39)Spermatogonial stem cell transplantation (SSCT) is an experimental technique for transfer of germline between donor and recipient males that could be used as a tool for biomedical research, preservation of endangered species, and dissemination of desirable genetics in food animal populations. To fully realize these potentials, recipient males must be devoid of endogenous germline but possess normal testicular architecture and somatic cell function capable of supporting allogeneic donor stem cell engraftment and regeneration of spermatogenesis. Here we show that male mice, pigs, goats, and cattle harboring knockout alleles of the <i>NANOS2</i> gene generated by CRISPR-Cas9 editing have testes that are germline ablated but otherwise structurally normal. In adult pigs and goats, SSCT with allogeneic donor stem cells led to sustained donor-derived spermatogenesis. With prepubertal mice, allogeneic SSCT resulted in attainment of natural fertility. Collectively, these advancements represent a major step toward realizing the enormous potential of surrogate sires as a tool for dissemination and regeneration of germplasm in all mammalian species.Proceedings of the National Academy of Sciences of the United States of AmericaDonor-derived spermatogenesis following stem cell transplantation in sterile <i>NANOS2</i> knockout males.PMC7533891BB/L007371/1IntramuralBBS/E/D/30002275BBS/E/D/20221658BBS/E/D/10002071BBS/E/D/20221656BB/M028313/12018-064782017-05436BB/M018342/1BB/N015339/1Park KETibary ALillico SRegouski MFan ZOatley MJPark CHTelugu BWaqas MSOatley JMWhitelaw BLiu YPolejaeva IARobbins CMiao DCiccarelli MLopez-Biladeau BGiassetti MIfalseDonor-derived spermatogenesis following stem cell transplantation in sterile <i>NANOS2</i> knockout males.Spermatogonial stem cell transplantation (SSCT) is an experimental technique for transfer of germline between donor and recipient males that could be used as a tool for biomedical research, preservation of endangered species, and dissemination of desirable genetics in food animal populations. To fully realize these potentials, recipient males must be devoid of endogenous germline but possess normal testicular architecture and somatic cell function capable of supporting allogeneic donor stem cell engraftment and regeneration of spermatogenesis. Here we show that male mice, pigs, goats, and cattle harboring knockout alleles of the <i>NANOS2</i> gene generated by CRISPR-Cas9 editing have testes that are germline ablated but otherwise structurally normal. In adult pigs and goats, SSCT with allogeneic donor stem cells led to sustained donor-derived spermatogenesis. With prepubertal mice, allogeneic SSCT resulted in attainment of natural fertility. Collectively, these advancements represent a major step toward realizing the enormous potential of surrogate sires as a tool for dissemination and regeneration of germplasm in all mammalian species.2020-01-01T00:00:00Z2020 Sep2024-11-19T15:49:40.655Z2020-10-29T09:25:30ZS-EPMC75338913292901210.1073/pnas.2010102117