Transgenic rhesus monkeys produced by gene transfer into early-cleavage-stage embryos using a simian immunodeficiency virus-based vector
2010
Niu, Yuyu | Yu, Yang | Bernat, Agnieszka | Yang, Shihua | He, Xiechao | Guo, Xiangyu | Chen, Dongliang | Chen, Yougchang | Ji, Shaohui | Si, Wei | Lv, Yongqin | Tan, Tao | Wei, Qiang | Wang, Hong | Shi, Lei | Guan, Jean | Zhu, Xuemei | Afanassieff, Marielle | Savatier, Pierre, Savatier | Zhang, Kang | Zhou, Qi | Ji, Weizhi | Chinese Academy of Sciences (CAS) | Kunming Institute of Zoology (KIZ) ; Chinese Academy of Sciences [Beijing] (CAS) | Institut cellule souche et cerveau (SBRI) ; Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL) ; Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM) | Université de Lyon | Kunming University of Science and Technology (KMUST) | University of California (UC) | Chinese Academy of Sciences [KSCX1-05-02, KSCX2-YW-R-47] ; Major State Basic Development Program [2006CB701505, 2007CB947701] ; National High Technology Research and Development Program [2006AA02A116, 2006AA02A101] ; National Key Technology RD Program [2007BAI33B00] ; China National Science Foundation [30701059, 30700425] ; R&D Infrastructure and Facility Development Program of Yunnan Province [2006PT08-2] ; Social Science and Technology Development Program of Yunnan Province [2007GH] ; National Science and Technology Major Project [2009ZX09501-028] ; Chinese National 985 Project ; US National Eye Institute/National Institutes of Health ; Veterans Administration ; Burroughs Wellcome Fund Clinical Scientist Award in Translational Research
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Show more [+] Less [-]English. 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.
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