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-cleavagestage 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. lentiviral vector | transgenesis

Modeling human neurodegenerative diseases in transgenic systems

“…A transgenic Huntington's disease model in rhesus monkeys (Macaca mulatta) has been prepared by lentiviral injection into the oocyte of polyglutamine-expanded Huntingtin (HTT) followed by fertilization by ICSI ). Transgenic rhesus monkeys have also been generated using a simian immunodeWciency virus (SIV)-based vector to infect early-cleavage stage embryos (Niu et al 2010). A major obstacle to generating transgenic non-human primates is the low eYciency of assisted reproductive technologies in producing oocytes and embryos suitable for genetic engineering as well as the technical challenges of embryonic stem cell derivation and cloning.…”

Production of Transgenic Nonhuman Primates

“…Due to the significant resources required for the creation of a germ line of transgenic NHPs (Chan et al, 2001;Yang et al, 2008;Sasaki et al, 2009;Niu et al, 2010;Sun et al, 2008), focal transgenesis (Kordower et al, 2000;Mittoux et al, 2000;Palfi et al, 2007), widely used in rodents, is also commonly used in NHPs as a short-term and relatively lowcost method for evaluating gene functions in higher primates. Although a wide range of genetic manipulation tools have been successfully used in rodents and other livestock species such as cattle and swine (Hauschild et al, 2011;HauschildQuintern et al, 2013;Tessanne et al, 2012;Golding et al, 2006), the production of transgenic NHPs has been limited to the overexpression approach (Chan et al, 2001;Yang et al, 2008;Sasaki et al, 2009;Niu et al, 2010;Sun et al, 2008) and primarily focused on dominant genetic diseases such as HD (Chan, 2004;Chan et al, 2001;Yang and Chan, 2011;Yang et al, 2008).…”

Prefrontal dysfunction and a monkey model of schizophrenia

“…It used to be quite diffi cult to perform genetic manipulation in monkeys, but at present, these techniques are rapidly developing in nonhuman primates [53,54] . Thus it will be possible to develop genetic monkey models for schizophrenia by genetic manipulation techniques in the near future.…”

“…Partial knockout of NRG1 in mice causes social interaction problems, reduced prepulse inhibition, and higher levels of spontaneous locomotion. These symptoms are reduced by clozapine [51] .It used to be quite diffi cult to perform genetic manipulation in monkeys, but at present, these techniques are rapidly developing in nonhuman primates [53,54] . Thus it will be possible to develop genetic monkey models for schizophrenia by genetic manipulation techniques in the near future.…”