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Monkeying about with transgenics

Nature Reviews Genetics volume 2, page 160 (2001)Cite this article

Chan et al. overcame the technical barrier to transgenic success in primates by adapting a vector that they had previously used to good effect in cattle. The key to this protocol is that the transgene-carrying retroviral vector is introduced into oocytes and not into embryos. Often retroviral vectors give rise to transgenic mosaics because they only integrate into dividing cells — cells in which the nuclear envelope is degraded during mitosis, so allowing the retroviral pre-integration complex access to the host cell DNA. Chan et al. therefore reasoned that, rather than targeting embryos, they would introduce their vector into metaphase II oocytes, which have no nuclear envelope. This timing should allow vector complexes to access the DNA. Additionally, as the genes are inserted before fertilization, the resulting offspring should not be mosaic.

So how successful was this strategy when transferred to rhesus monkeys? The authors injected their green fluorescent protein-encoding retroviral vector — either under the control of the cytomegalovirus early promoter or the human elongation factor-1 alpha promoter — into 224 mature rhesus oocytes. Six hours later, they fertilized them. Of these 224 fertilized oocytes, 126 developed to 4-cell stage embryos, 40 of which were selected by their morphology for transfer, in pairs, to 20 surrogate mothers. Five pregnancies resulted, three of which produced healthy males. One pregnancy miscarried fraternal twins mid-gestation, possibly because rhesus monkeys rarely sustain twin pregnancies.

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References

ORIGINAL RESEARCH PAPER

  1. Chan, A. W. S. et al. Transgenic monkeys produced by retroviral gene transfer into mature oocytes. Science 291, 309–312 ( 2001) [Contents page]

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FURTHER READING

  1. Chan, A. W. et al. Transgenic cattle produced by reverse-transcribed gene transfer in oocytes. Proc. Natl Acad. Sci. USA 95, 14028–14033 (1998) http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9826647&dopt=Abstract

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Authors
  1. Jane Alfred
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Alfred, J. Monkeying about with transgenics. Nat Rev Genet 2, 160 (2001). https://doi.org/10.1038/35056000

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