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Complementation of placental defects and embryonic lethality by trophoblast-specific lentiviral gene transfer

Nature Biotechnology volume 25, pages 233–237 (2007)Cite this article

Abstract

Placental dysfunction underlies many complications during pregnancy, and better understanding of gene function during placentation could have considerable clinical relevance. However, the lack of a facile method for placenta-specific gene manipulation has hampered investigation of placental organogenesis and the treatment of placental dysfunction. We showed previously that transduction of fertilized mouse eggs with lentiviral vectors leads to transgene expression in both the fetus and the placenta1,2. Here we report placenta-specific gene incorporation by lentiviral transduction of mouse blastocysts after removal of the zona pellucida. All of the placentas analyzed, but none of the fetuses, were transgenic. Application of this method substantially rescued mice deficient in Ets2, Mapk14 (also known as p38α) and Mapk1 (also known as Erk2) from embryonic lethality caused by placental defects. Ectopic expression of Mapk11 also complemented Mapk14 deficiency during placentation.

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Figure 1: Lentiviral vector transduction of blastocysts results in placenta-specific gene expression.
Figure 2: Restoration of placental defects and embryonic lethality in Ets2−/− mice by placenta-specific gene expression.
Figure 3: Functional complementation of Mapk14 with Mapk11during placentation.

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Acknowledgements

We thank H.A. Popiel for critical reading of the manuscript, and Y. Maruyama, A. Kawai, Y. Esaki, M. Tanaka and Y. Koreeda for technical assistance. Rat anti-GFP antibody was kindly provided by Fujita of Mitsubishi-Kasei Institute of Life Sciences. This research was supported in part by Uehara Memorial Foundation, Japan Chemical Industry Association (JCIA), and MEXT of Japan.

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Authors and Affiliations

  1. Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka, 565-0871, Japan

    Yuka Okada, Yuko Ueshin, Ayako Isotani, Tomoko Saito-Fujita, Masaru Okabe & Masahito Ikawa

  2. Graduate School of Pharmaceutical Sciences, Osaka University, 3-1 Yamada-oka, Suita, Osaka, 565-0871, Japan

    Yuko Ueshin, Ayako Isotani & Masaru Okabe

  3. School of Medicine, Mie University, 2-174 Edobashi, Tsu, 514-8507, Mie, Japan

    Hisako Nakashima, Kazushi Kimura, Akira Mizoguchi, Masatsugu Oh-hora, Yoshiko Mori & Masato Ogata

  4. The Burnham Institute for Medical Research, 10901 North Torrey Pines Road, La Jolla, 92037, California, USA

    Robert G Oshima

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Contributions

Y.O. performed research, analyzed data and wrote the manuscript; Y.U. performed research; A.I. performed tetraploid complementation; T.S.F. performed quantitative real-time RT-PCR; H.N. performed histological analysis; K.K. performed histological analysis; A.M. performed histological analysis and provided useful discussions; M.O. provided p38α mutant mice; Y.M. provided useful reagents; M.O. provided p38α and Mapk1 mutant mice and useful discussions; R.G.O. provided Ets2 mutant mice and useful discussions; M.O. provided useful discussions and wrote manuscript; M.I. designed, coordinated and performed research, analyzed data and wrote manuscript.

Corresponding author

Correspondence to Masahito Ikawa.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Figure 1

Chromosomal localization of the lentiviral transgenes after transduction of a zona-free blastocyst. (PDF 83 kb)

Supplementary Figure 2

E12.5 embryos transduced with LV-EGFP lentiviral vector by blastocoel injection (top) or co-incubation with zona-free blastocysts (bottom). (PDF 331 kb)

Supplementary Figure 3

Average body and placenta weight of Ets2 mutant mice at birth after lentiviral vector transduction. (PDF 86 kb)

Supplementary Table 1

Genotypes of E13.5 embryos after lentiviral vector transduction. (PDF 31 kb)

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Okada, Y., Ueshin, Y., Isotani, A. et al. Complementation of placental defects and embryonic lethality by trophoblast-specific lentiviral gene transfer. Nat Biotechnol 25, 233–237 (2007). https://doi.org/10.1038/nbt1280

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