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Transgenic rhesus monkeys produced by gene transfer into early-cleavage–stage embryos using a simian immunodeficiency virus-based vector
Yuyu Niua,b,c,d,1, Yang Yue,1, Agnieszka Bernatf,g,h,1, Shihua Yanga,b,i, Xiechao Hea,b,c,d, Xiangyu Guoa,b,c,d, Dongliang Chena,b,c,d, Yongchang Chena,b,c,d, Shaohui Jia,b,c, Wei Sia,b,c,d, Yongqin Lva,b,c,d, Tao Tana,b,c,d, Qiang Weia,b,c,d, Hong Wanga,b,c,d, Lei Shia,b,c,d, Jean Guanj,k,l,m, Xuemei Zhul,m, Marielle Afanassieffe,f,g, Pierre Savatierf,g,h,2, Kang Zhangj,k,l,m,2, Qi Zhoue,2, and Weizhi Jia,b,c,d,2
aKunming Primate Research Center and
bKunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, People's Republic of China;
cKunming Biomed International and
dNational Engineering Research Center of Biomedicine and Animal Science, Kunming 650051, People's Republic of China;
eInstitute of Zoology, Chinese Academy of Sciences, Beijing 100860, People's Republic of China;
fInstitut National de la Santé et de la Recherche Médicale, U846, and
gPrimaStem Stem Cell and Brain Research Institute, Bron 69500, France;
hUniversité de Lyon, Lyon 69003, France;
iBiotechnology Research Center, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650093, People's Republic of China;
jCenter for Molecular Medicine and Department of Ophthalmology, West China Hospital and
kSichuan University, Chengdu 610065, People's Republic of China; and
lInstitute for Genomic Medicine and
mShiley Eye Center, University of California, San Diego, La Jolla, CA 92093
The development of transgenic technologies in monkeys is important for creating valuable animal models of human physiology so that the etiology of diseases can be studied and potential therapies for their amelioration may be developed. However, the efficiency of producing transgenic primate animals is presently very low, and there are few reports of success. We have developed an improved methodology for the production of transgenic rhesus monkeys, making use of a simian immunodeficiency virus (SIV)-based vector that encodes EGFP and a protocol for infection of early-cleavage–stage embryos. We show that infection does not alter embryo development. Moreover, the timing of infection, either before or during embryonic genome activation, has no observable effect on the level and stability of transgene expression. Of 70 embryos injected with concentrated virus at the one- to two-cell stage or the four- to eight-cell stage and showing fluorescence, 30 were transferred to surrogate mothers. One transgenic fetus was obtained from a fraternal triple pregnancy. Four infant monkeys were produced from four singleton pregnancies, of which two expressed EGFP throughout the whole body. These results demonstrate the usefulness of SIV-based lentiviral vectors for the generation of transgenic monkeys and improve the efficiency of transgenic technology in nonhuman primates.
