Reproduction Abstracts

Introduction: Nonhuman primates are considerered valuable human disease models. There are not many reports about transgenic monkey and especially no report about GFP cynomolgus monkey. Here, we report for the first time generation of whole body GFP expressing cynomolgus monkey.

Methods: We constructed a lentiviral vector carries GFP under control CAG promoter. To generate a transgenic cynomolgus monkey expressed GFP uniformly, we compared injection timing of lentiviral vector. Lentivirus were injected perivitelline space of oocytes befor 4 h of fertilization via intracytoplasmic sperm injection (ICSI) (PreI)or 24 h after fertilization via ICSI (PostI).

Results and Discussion: Transfer of five PostI embryos into three recipients resulted in two pregnancies, one of which ended in miscarriage of twins on 92 of gestation. Although one of the twins (#1) showed no detectable fluorescence, the other (#2) showed strong fluorescence in the face, skin, placenta and brain. Partial GFP expression in the peripheral blood and fibroblast established from tail were observed. An offspring (named PreI Tg #1) from the PreI embryo was born successfully and showed strong fluorescence in the face and placenta. Amnion of PreI Tg #1 monkey showed the uniform GFP expression at the cellular level. Another offspring (named PreI Tg #2) was born, but died 3 days after birth. Uniform GFP expression in the peripheral blood and fibroblast established from tail were observed. Collectively, these results demonstrated that the GFP cynomolgus monkey created by PreI technique expressed GFP uniformly.