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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Animal Biosciences & Biotechnology Laboratory » Research » Publications at this Location » Publication #334806
Title: Generation of germline ablated male pigs by CRISPR/Cas9 editing of the NANOS2 gene

Author
item KI-EUN, PARK - University Of Maryland
item KAUCHER, AMY - Washington State University
item Powell, Anne
item WAGAS, MUHAMAD SALMAN - Washington State University
item SANDMAIER, SHELLEY E.S. - University Of Maryland
item OATLEY, MELISSA - Washington State University
item PARK, CHI-HUN - University Of Maryland
item TIBARY, AHMED - Washington State University
item Donovan, David
item Blomberg, Le Ann
item LILLICO, SIMON - University Of Edinburgh
item WHITELAW, BRUCE - University Of Edinburgh
item MILEHAM, ALAN - Genus
item TELUGU, BHANU - University Of Maryland
item OATLEY, JON - Washington State University

Submitted to: Nature Communications
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/2/2016
Publication Date: 1/10/2017
Citation: Ki-Eun, P., Kaucher, A., Powell, A.M., Wagas, M., Sandmaier, S., Oatley, M.J., Park, C., Tibary, A., Donovan, D.M., Blomberg, L., Lillico, S., Whitelaw, B., Mileham, A., Telugu, B., Oatley, J.M. 2017. Generation of germline ablated male pigs by CRISPR/Cas9 editing of the NANOS2 gene. Nature Communications. doi: 10.1038/srep40176.

Interpretive Summary: To increase breeding efficiency in livestock, it is important to understand the genes that control development of the reproductive system. In mice the NANOS2 gene has been shown (via gene knockout technology) to be important for the development of sperm but is not essential for female reproductive organs. This information was not known for pig until this report in which gene editing technology allowed for the production of NANOS2 knockout pigs. The pig matches the mouse in phenotype, with the female NANOS2 knockout animals being fertile, while the male knockouts are sterile. This work will help scientists world-wide to recognize the increased value of the pig as a good model organism for studying reproduction and in particular, the role of NANOS2 in sperm production. It is also a goal of this work to eventually use the NANOS2 knockout male pigs for spermatagonial stem cell transplant and thus expand the source of elite animals that can make this elite sperm, making these desirable germplasm more readily available to the pig farmer.

Technical Abstract: Genome editing tools have revolutionized the generation of genetically modified animals including livestock. In particular, the domestic pig is a proven model of human physiology and an agriculturally important species. In this study, we utilized the CRISPR/Cas9 system to edit the NANOS2 gene in pig embryos to generate offspring with mono-allelic and bi-allelic mutations. We found that NANOS2 knockout pigs phenocopy knockout mice with male specific germline ablation but other aspects of testicular development are normal. Moreover, male pigs with one intact NANOS2 allele and female knockout pigs are fertile. From an agriculture perspective, NANOS2 knockout male pigs are expected to serve as an ideal surrogate for transplantation of donor spermatogonial stem cells to expand the availability of gametes from genetically desirable sires.