Transgenic rhesus monkeys produced by gene transfer into early-cleavage-stage embryos using a simian immunodeficiency virus-based vector.
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.
Niu
Yuyu
Y
Kunming Primate Research Center and Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, People's Republic of China.
Yu
Yang
Y
Bernat
Agnieszka
A
Yang
Shihua
S
He
Xiechao
X
Guo
Xiangyu
X
Chen
Dongliang
D
Chen
Yongchang
Y
Ji
Shaohui
S
Si
Wei
W
Lv
Yongqin
Y
Tan
Tao
T
Wei
Qiang
Q
Wang
Hong
H
Shi
Lei
L
Guan
Jean
J
Zhu
Xuemei
X
Afanassieff
Marielle
M
Savatier
Pierre
P
Zhang
Kang
K
Zhou
Qi
Q
Ji
Weizhi
W
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D015139:Blotting, Southern; D002970:Cleavage Stage, Ovum; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D007150:Immunohistochemistry; D008253:Macaca mulatta / Q000235:genetics*; D023421:Models, Animal; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D015302:Simian Immunodeficiency Virus
D017931:DNA Primers; D005456:Fluorescent Dyes; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins