title: Transgenic rhesus monkeys produced by gene transfer into early-cleavage-stage embryos using a simian immunodeficiency virus-based vector.
abstract: 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.
authors.affiliation: Kunming Primate Research Center and Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, People's Republic of China.
authors.forename: Yuyu
authors.initials: Y
authors.lastname: Niu
authors.forename: Yang
authors.initials: Y
authors.lastname: Yu
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Bernat
authors.forename: Shihua
authors.initials: S
authors.lastname: Yang
authors.forename: Xiechao
authors.initials: X
authors.lastname: He
authors.forename: Xiangyu
authors.initials: X
authors.lastname: Guo
authors.forename: Dongliang
authors.initials: D
authors.lastname: Chen
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Chen
authors.forename: Shaohui
authors.initials: S
authors.lastname: Ji
authors.forename: Wei
authors.initials: W
authors.lastname: Si
authors.forename: Yongqin
authors.initials: Y
authors.lastname: Lv
authors.forename: Tao
authors.initials: T
authors.lastname: Tan
authors.forename: Qiang
authors.initials: Q
authors.lastname: Wei
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Lei
authors.initials: L
authors.lastname: Shi
authors.forename: Jean
authors.initials: J
authors.lastname: Guan
authors.forename: Xuemei
authors.initials: X
authors.lastname: Zhu
authors.forename: Marielle
authors.initials: M
authors.lastname: Afanassieff
authors.forename: Pierre
authors.initials: P
authors.lastname: Savatier
authors.forename: Kang
authors.initials: K
authors.lastname: Zhang
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhou
authors.forename: Weizhi
authors.initials: W
authors.lastname: Ji
mesh_terms: 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
chemical_list: D017931:DNA Primers; D005456:Fluorescent Dyes; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins