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High-frequency generation of viable mice from engineered bi-maternal embryos

Nature Biotechnology volume 25, pages 1045–1050 (2007)Cite this article

Abstract

Mammalian development to adulthood typically requires both maternal and paternal genomes, because genomic imprinting places stringent limitations on mammalian development, strictly precluding parthenogenesis1,2. Here we report the generation of bi-maternal embryos that develop at a high success rate equivalent to the rate obtained with in vitro fertilization of normal embryos. These bi-maternal mice developed into viable and fertile female adults. The bi-maternal embryos, distinct from parthenogenetic or gynogenetic conceptuses, were produced by the construction of oocytes from fully grown oocytes and nongrowing oocytes that contain double deletions in the H19 differentially methylated region (DMR) and the Dlk1-Dio3 intergenic germline–derived DMR. The results provide conclusive evidence that imprinted genes regulated by these two paternally methylated imprinting-control regions are the only paternal barrier that prevents the normal development of bi-maternal mouse fetuses to term.

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Figure 1: The bi-maternal mice derived from the ngΔDouble/fg embryos.
Figure 2: Graphical representations of the expression of the two sets of imprinted genes in the bi-maternal fetuses.
Figure 3: Global gene expression analysis using microarray.

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References

  1. Surani, M.A., Barton, S.C. & Norris, M.L. Development of reconstituted mouse eggs suggests imprinting of the genome during gametogenesis. Nature 308, 548–550 (1984).

    Article  CAS  Google Scholar 

  2. McGrath, J. & Solter, D. Completion of mouse embryogenesis requires both the maternal and paternal genomes. Cell 37, 179–183 (1984).

    Article  CAS  Google Scholar 

  3. Kono, T., Obata, Y., Yoshimzu, T., Nakahara, T. & Carroll, J. Epigenetic modifications during oocyte growth correlates with extended parthenogenetic development in the mouse. Nat. Genet. 13, 91–94 (1996).

    Article  CAS  Google Scholar 

  4. Obata, Y. et al. Disruption of primary imprinting during oocyte growth leads to the modified expression of imprinted genes during embryogenesis. Development 125, 1553–1560 (1998).

    CAS  PubMed  Google Scholar 

  5. Obata, Y. & Kono, T. Maternal primary imprinting is established at a specific time for each gene throughout oocyte growth. J. Biol. Chem. 277, 5285–5289 (2002).

    Article  CAS  Google Scholar 

  6. Kono, T. et al. Birth of parthenogenetic mice that can develop to adulthood. Nature 428, 860–864 (2004).

    Article  CAS  Google Scholar 

  7. Tremblay, K.D., Saam, J.R., Ingram, R.S., Tilghman, S.M. & Bartolomei, M.S. A paternal-specific methylation imprint marks the alleles of the mouse H19 gene. Nat. Genet. 9, 407–413 (1995).

    Article  CAS  Google Scholar 

  8. Pearsall, R.S. et al. A direct repeat sequence at the Rasgrf1 locus and imprinted expression. Genomics 55, 194–201 (1999).

    Article  CAS  Google Scholar 

  9. Lin, S.P. et al. Asymmetric regulation of imprinting on the maternal and paternal chromosomes at the Dlk1-Gtl2 imprinted cluster on mouse chromosome 12. Nat. Genet. 35, 97–102 (2003).

    Article  CAS  Google Scholar 

  10. Leighton, P.A., Ingram, R.S., Eggenschwiler, J., Efstratiadis, A. & Tilghman, S.M. Disruption of imprinting caused by deletion of the H19 gene region in mice. Nature 375, 34–39 (1995).

    Article  CAS  Google Scholar 

  11. Clapcott, S.J., Peters, J., Orban, P.C., Brambilla, R. & Graham, C.F. Two ENU-induced mutations in Rasgrf1 and early mouse growth retardation. Mamm. Genome 14, 495–505 (2003).

    Article  CAS  Google Scholar 

  12. Brambilla, R. et al. A role for the Ras signalling pathway in synaptic transmission and long-term memory. Nature 390, 281–286 (1997).

    Article  CAS  Google Scholar 

  13. Kawahara, M., Wu, Q., Yaguchi, Y., Ferguson-Smith, A.C. & Kono, T. Complementary roles of genes regulated by two paternally methylated imprinted regions on chromosomes 7 and 12 in mouse placentation. Hum. Mol. Genet. 15, 2869–2879 (2006).

    Article  CAS  Google Scholar 

  14. Zeng, F. & Schultz, R.M. RNA transcript profiling during zygotic gene activation in the preimplantation mouse embryo. Dev. Biol. 283, 40–57 (2005).

    Article  CAS  Google Scholar 

  15. Su, Y.Q. et al. Selective degradation of transcripts during meiotic maturation of mouse oocytes. Dev. Biol. 302, 104–117 (2007).

    Article  CAS  Google Scholar 

  16. Fielder, P.J., Thordarson, G., English, A., Rosenfeld, R.G. & Talamantes, F. Expression of a lactogen-dependent insulin-like growth factor-binding protein in cultured mouse mammary epithelial cells. Endocrinology 131, 261–267 (1992).

    Article  CAS  Google Scholar 

  17. Delany, A.M., Rydziel, S. & Canalis, E. Autocrine down-regulation of collagenase-3 in rat bone cell cultures by insulin-like growth factors. Endocrinology 137, 4665–4670 (1996).

    Article  CAS  Google Scholar 

  18. Stickens, D. et al. Altered endochondral bone development in matrix metalloproteinase 13-deficient mice. Development 131, 5883–5895 (2004).

    Article  CAS  Google Scholar 

  19. Moon, Y.S. et al. Mice lacking paternally expressed Pref-1/Dlk1 display growth retardation and accelerated adiposity. Mol. Cell. Biol. 22, 5585–5592 (2002).

    Article  CAS  Google Scholar 

  20. Sakajiri, S. et al. Dlk1 in normal and abnormal hematopoiesis. Leukemia 19, 1404–1410 (2005).

    Article  CAS  Google Scholar 

  21. Ostermeier, G.C., Miller, D., Huntriss, J.D., Diamond, M.P. & Krawetz, S.A. Reproductive biology: delivering spermatozoan RNA to the oocyte. Nature 429, 154 (2004).

    Article  CAS  Google Scholar 

  22. Kono, T., Sotomaru, Y., Katsuzawa, Y. & Dandolo, L. Mouse parthenogenetic embryos with monoallelic H19 expression can develop to day 17.5 of gestation. Dev. Biol. 243, 294–300 (2002).

    Article  CAS  Google Scholar 

  23. Wu, Q. et al. Regulated expression of two sets of paternally imprinted genes is necessary for mouse parthenogenetic development to term. Reproduction 131, 481–488 (2006).

    Article  CAS  Google Scholar 

  24. Seitz, H. et al. Imprinted microRNA genes transcribed antisense to a reciprocally imprinted retrotransposon-like gene. Nat. Genet. 34, 261–262 (2003).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Shirley Tilghman, Princeton University, for the gift of mutant mice and Hidehiko Ogawa, Yayoi Obata and Yusuke Sotomaru for discussions. This work was supported by grants from the Bio-oriented Technology Research Advancement Institution of Japan, the Ministry of Education, Science, Culture and Sports of Japan, and the UK Biotechnology and Biological Sciences Research Council.

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Author notes

  1. Qiong Wu

    Present address: Present address: Department of Life Science and Engineering Harbin Institute of Technology, No.92, West Da-Zhi Street, Harbin, Heilongjiang.,

Authors and Affiliations

  1. Department of BioScience Tokyo University of Agriculture, Setagaya-ku, Tokyo, 156-8502, Japan

    Manabu Kawahara, Qiong Wu, Nozomi Takahashi, Shinnosuke Morita, Kaori Yamada & Tomohiro Kono

  2. Bio-oriented Technology Research Advancement Institution (BRAIN), Minato-ku, Tokyo, 105-0001, Japan

    Manabu Kawahara, Qiong Wu & Tomohiro Kono

  3. Department of Physiology, Development & Neuroscience, University of Cambridge, Downing St., Cambridge, CB2 3DY, UK

    Mitsuteru Ito & Anne C Ferguson-Smith

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  1. Manabu Kawahara
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Contributions

M.K. and T.K. designed the experiments, analyzed the data and wrote the manuscript. M.K., Q.W., N.T., S.M., K.Y. and M.I. performed experiments. A.C.F.-S. provided mutant mice and wrote the manuscript.

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Correspondence to Tomohiro Kono.

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The authors declare no competing financial interests.

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Kawahara, M., Wu, Q., Takahashi, N. et al. High-frequency generation of viable mice from engineered bi-maternal embryos. Nat Biotechnol 25, 1045–1050 (2007). https://doi.org/10.1038/nbt1331

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