Generation of transgenic cynomolgus monkeys that express green fluorescent protein throughout the whole body

全身表达绿色荧光蛋白的转基因食蟹猴的产生

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作者:Yasunari Seita, Tomoyuki Tsukiyama, Chizuru Iwatani, Hideaki Tsuchiya, Jun Matsushita, Takuya Azami, Junko Okahara, Shinichiro Nakamura, Yoshitaka Hayashi, Seiji Hitoshi, Yasushi Itoh, Takeshi Imamura, Masaki Nishimura, Ikuo Tooyama, Hiroyuki Miyoshi, Mitinori Saitou, Kazumasa Ogasawara, Erika Sasak

Abstract

Nonhuman primates are valuable for human disease modelling, because rodents poorly recapitulate some human diseases such as Parkinson's disease and Alzheimer's disease amongst others. Here, we report for the first time, the generation of green fluorescent protein (GFP) transgenic cynomolgus monkeys by lentivirus infection. Our data show that the use of a human cytomegalovirus immediate-early enhancer and chicken beta actin promoter (CAG) directed the ubiquitous expression of the transgene in cynomolgus monkeys. We also found that injection into mature oocytes before fertilization achieved homogenous expression of GFP in each tissue, including the amnion, and fibroblasts, whereas injection into fertilized oocytes generated a transgenic cynomolgus monkey with mosaic GFP expression. Thus, the injection timing was important to create transgenic cynomolgus monkeys that expressed GFP homogenously in each of the various tissues. The strategy established in this work will be useful for the generation of transgenic cynomolgus monkeys for transplantation studies as well as biomedical research.

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