The Non-human Primate Facility of ION was established in August 2009, which take cynomolgus monkey, rhesus monkey as the basic animal model to establish transgenic monkey and do relevant research work. Cynomolgus monkey and rhesus monkey are the main non-human primate models for our experiment. They play a very important role in the field of life science and biomedicine. With the establishment of monkey assisted reproductive technology and lentiviral vector transfection and the development of genome targeting methods (ZFN, TALEN, CRISPR/Cas9) , scientists have successfully obtained transgenic monkeys and gene modified monkeys. Our laboratory is dedicated to the construction of non-human primate gene modified models and the study of related reproductive, physiology, assisted reproduction, gene transfection, gene modification and stem cell technology.
1, Study on reproductive physiology and reproductive acceleration in non-human primates
Rhesus monkeys and cynomolgus monkeys are highly similar to humans in reproductive physiology, they are both monotocous species with the menstrual cycle. Assisted reproductive technology is the basis for constructing non-human primate gene modified model. In 2008, we established high efficient techniques for embryo construction and embryo transfer, and obtained the first test-tube cynomolgus monkeys in China. On this basis, we are cooperating with the assisted reproductive center of Shanghai No.9 People's Hospital, to make a study on the regulation mechanism of neural and endocrine of primate reproduction, we are committed to optimizing the ovulation strategies of non-human primates, and improving the embryo quality and pregnancy outcome.
A major obstacle to the use of nonhuman primates as model animals is their long reproductive cycle. The sexual maturity of the rhesus monkeys and cynomolgus monkeys is more than 4 years. In the previous work, we established a sperm maturation accelerated technique based on testicular xenografting, which successfully shortened the reproductive duration of cynomolgus monkeys from 4.5 years to 2.5 years, and obtained healthy cynomolgus monkeys offspring. We will further optimize the accelerated maturation of monkeys based on the study of the regulation mechanisms of reproductive hormones in primates.
2, The construction of non-human primate models and the optimization of the technology
We have established the lentiviral injection technology, and the gene modified （TALEN, CRISPR/Cas9） methods , and successfully obtained MeCP2, Prrt2, Fmr1 and Bmal1 and other gene modified founder monkeys. But most of these founder monkeys are mosaic. To solve this problem, we succeeded in using a combination of multiple sgRNA injection, and get fully mutated Prrt2 and Bmal1 monkeys. Based on the study, we will continue to optimize the technology of genome modification in monkeys, and cooperate with the research group in our institute to construct and study on the transgene or gene knockout non-human primate models related to the nerve disease.
3, Study on new gene modification technology for non-human primate models
In the construction of mouse model, we can also use ES-injection, spermatogonial stem cells injection, somatic cell nuclear transfer and haploid stem cell injection to obtain mouse models. Although these methods are not as simple and easy operation as CRISPR/Cas9 method, but we can get animal models without mosaicism by these methods. In the previous work, we have tried and established the parthenogenetic haploid stem cell line of cynomolgus monkeys, and we also established the embryonic stem cells from rhesus monkeys and cynomolgus monkeys. On this basis, we will continue to study the totipotent of monkey ES cells, the semi cloning and cloning of monkey.
