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Possible mechanism of gene transfer into early to mid-gestational mouse fetuses by tail vein injection

Gene Therapy volume 9, pages 1529–1541 (2002)Cite this article

Abstract

Our aim is to develop a simple gene transfer method into egg cylinder and mid-gestational murine embryos. We examined whether plasmid/lipid complexes injected into the tail veins of pregnant transgenic mice can be transferred to fetuses at E 4.5–13.5. When pregnant CETZ-17 mice carrying a transgene consisting of a ubiquitous promoter, floxed EGFP/CAT and the LacZ gene, were injected with a Cre expression vector DNA/lipid complex, Cre-mediated excision of the transgenes, as evaluated by X-gal staining, occurred in 10–50% of fetuses treated at E 11.5–13.5. Although younger embryos remained unstained, PCR analysis revealed low levels of the Cre vector DNA and recombined transgene. To examine the fate of a solution given intravenously, we injected trypan blue or fluorescence-labeled plasmid DNA/lipid complexes into females at E 5.5–11.5 and E 6.5, respectively. Both collected in the visceral endoderm (VE) lineage, but were undetectable in the embryo proper. These findings suggest that substances in maternal blood are delivered to post-implantation embryos via cells of the VE lineage and placenta, but that most are trapped in the VE. If significantly improved, gene transfer to fetuses by injection into the maternal circulation may become a promising tool in fetal gene therapy and embryological studies.

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Acknowledgements

We thank Drs Kimi Araki (Kumamoto University) and Jun-ichi Miyazaki (Osaka University) for providing us with CAG-Cre transgenic mice and pCAG-CAT-lacZ plasmid, respectively, and Dr Osamu Tanaka (Tokai University) for discussion. MS and MK were supported by Grants-in-Aid from the Ministry of Education, Science, Sports and Culture, Japan.

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Authors and Affiliations

  1. Division of Molecular Life Science, School of Medicine, Tokai University, Isehara, Japan

    N Kikuchi, M Ohtsuka & M Kimura

  2. Division of Nephrology and Metabolism, School of Medicine, Tokai University, Isehara, Japan

    S Nakamura

  3. The Institute of Medical Sciences, Tokai University, Isehara, Japan

    M Sato

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  1. N Kikuchi
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  2. S Nakamura
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Kikuchi, N., Nakamura, S., Ohtsuka, M. et al. Possible mechanism of gene transfer into early to mid-gestational mouse fetuses by tail vein injection. Gene Ther 9, 1529–1541 (2002). https://doi.org/10.1038/sj.gt.3301818

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