Non-viral generation of transgenic non-human primates via the piggyBac transposon system
Non-human primates, such as cynomolgus monkeys, are invaluable experimental models for understanding human biology and disease. Their close genetic relationship to humans makes them essential for studying fundamental human developmental processes and disease progression. Although lentiviral methods for generating transgenic monkeys exist, several inherent technical difficulties limit their utility. To solve this problem, here we establish a non-viral method for generating transgenic cynomolgus monkeys using the piggyBac transposon system. After optimizing our protocol in mice, we show that the co-injection of piggyBac components with sperm into metaphase II-stage oocytes successfully generates transgenic monkeys expressing transgenes throughout their whole bodies. Transgene expression is observed in all examined tissue types, including germ cells, although the levels of expression vary. Insertion analysis further confirms the successful integration of the transgene. We propose that our method will be a practical non-viral protocol for generating transgenic non-human primates.
Authors
- Ichiro Terakado
- Mitinori Saitou (ORCID: https://orcid.org/0000-0002-2895-6798)
- Takuya Yamamoto (ORCID: https://orcid.org/0000-0002-0022-3947)
- Masataka Nakaya
- Ikuo Kawamoto
- Ai Mieda
- Setsuko Tsukiyama-Fujii
- Tomonori Nakamura (ORCID: https://orcid.org/0000-0002-0173-0780)
- Hideaki Tsuchiya
- Takahiro Nakagawa (ORCID: https://orcid.org/0009-0003-5045-1336)
- Iori Itagaki
- Takafumi Ichikawa (ORCID: https://orcid.org/0000-0002-0490-4176)
- Tomoyuki Tsukiyama (ORCID: https://orcid.org/0000-0002-2657-7719)
- Taro Tsujimura (ORCID: https://orcid.org/0000-0002-3281-0150)
- Chizuru Iwatani
- Shoko Tarumoto
Institutions
- Shiga University of Medical Science (JP)
- Kyoto University (JP)
- Japan Agency for Medical Research and Development (JP)
- Advanced Science, Technology & Management Research Institute of Kyoto (JP)
- RIKEN Center for Advanced Intelligence Project (JP)
- Corporation for Production and Research of Laboratory Primates (JP)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-03-24
- DOI
- https://doi.org/10.1038/s41467-025-57365-w
- Citations
- 1
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 0.81
Funders
- Takeda Science Foundation
- Mochida Memorial Foundation for Medical and Pharmaceutical Research
- Japan Society for the Promotion of Science