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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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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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