Proceedings of the National Academy of Sciences of the United States of America ( IF 9.412 ) Pub Date : 2020-09-14 , DOI: 10.1073/pnas.2010102117 Michela Ciccarelli, Mariana I. Giassetti, Deqiang Miao, Melissa J. Oatley, Colton Robbins, Blanca Lopez-Biladeau, Muhammad Salman Waqas, Ahmed Tibary, Bruce Whitelaw, Simon Lillico, Chi-Hun Park, Ki-Eun Park, Bhanu Telugu, Zhiqiang Fan, Ying Liu, Misha Regouski, Irina A. Polejaeva, Jon M. Oatley
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 NANOS2 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.