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Generation of transgenic non-human primates with germline transmission

Abstract

The common marmoset (Callithrix jacchus) is increasingly attractive for use as a non-human primate animal model in biomedical research. It has a relatively high reproduction rate for a primate, making it potentially suitable for transgenic modification. Although several attempts have been made to produce non-human transgenic primates, transgene expression in the somatic tissues of live infants has not been demonstrated by objective analyses such as polymerase chain reaction with reverse transcription or western blots. Here we show that the injection of a self-inactivating lentiviral vector in sucrose solution into marmoset embryos results in transgenic common marmosets that expressed the transgene in several organs. Notably, we achieved germline transmission of the transgene, and the transgenic offspring developed normally. The successful creation of transgenic marmosets provides a new animal model for human disease that has the great advantage of a close genetic relationship with humans. This model will be valuable to many fields of biomedical research.

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Figure 1: Self-inactivating lentiviral vector-derived EGFP transgenic marmosets.
Figure 2: Transgene insertions in several infant tissues.
Figure 3: Transgene transcription and expression in several infant tissues.
Figure 4: Germline transmission of the transgene.

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Acknowledgements

We thank F. Toyoda, S. Ohba, T. Inoue, Y. Sawada and M. Yokoyama for technical assistance with the animal experiments and care. E.S. is an associate professor of the Global COE program for human metabolomic systems biology assigned to Keio University. This study was also supported by the Global COE program for Education and Research Centre for Stem Cell Medicine from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), the Japanese Government to Keio University. This study was also supported by funds from Solution-Oriented Research for Science and Technology (SORST) of the Japan Science and Technology Agency and grants from MEXT to H.O. and from Special Coordination Funds for Promoting Science and Technology of MEXT to S.H.

Author Contributions E.S. designed the experiments, conducted the project, and wrote the paper. A.S., Y.S., T.E., I.T. and R.H. assisted in embryological technique development. K.H., R.O. and M.K. developed surgical techniques for embryo collection and transfer. H.S., C.K. and C.Y. performed or assisted with the real-time PCR and parentage evaluation test. S.S. and T.M. assisted with the Southern blot analysis and tissue collection. M.I. raised the anti-marmoset CD45 antibody. R.I. performed the FACS analysis, and K.K. performed the immunohistochemical analysis. H.M. provided the lentiviral vectors. Y.T., H.O., S.H., N.T. and T.N. designed the project, and H.O., S.H. and N.T. also participated in writing the paper. The whole project was supervised by E.S. and H.O.

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Correspondence to Erika Sasaki or Hideyuki Okano.

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Sasaki, E., Suemizu, H., Shimada, A. et al. Generation of transgenic non-human primates with germline transmission. Nature 459, 523–527 (2009). https://doi.org/10.1038/nature08090

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