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Fetal gene transfer by transuterine injection of cationic liposome–DNA complexes

Nature Biotechnology volume 17, pages 1188–1192 (1999)Cite this article

Abstract

In utero injection of cationic liposome–DNA complexes (CLDCs) containing chloramphenicol acetyltransferase, β-galactosidase (β-gal), or human granulocyte colony-stimulating factor (hG-CSF) expression plasmids produced high-level gene expression in fetal rats. Tissues adjacent to the injection site exhibited the highest levels of gene expression. Chloramphenicol acetyltransferase expression persisted for at least 14 days and was reexpressed following postnatal reinjection of CLDCs. Intraperitoneal administration of the hG-CSF gene produced high serum hG-CSF levels. X-gal staining demonstrated widespread β-gal expression in multiple fetal tissues and cell types. No toxic or inflammatory responses were observed, nor was there evidence of fetal–maternal or maternal–fetal gene transfer, suggesting that CLDCs may provide a useful alternative to viral vectors for in utero gene transfer.

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Figure 1: (A) Chloramphenicol acetyltransferase activity in liver and spleen extracts from fetal rats after receiving a single i.p. injection of either 12.5 μg (0.25 μg/μl) or 50 μg (1.0 μg/μl) of a CMV-CAT expression plasmid, either complexed to 50 or 200 nmol of DOTIM:cholesterol liposomes, respectively, or as DNA alone.
Figure 2: Chloramphenicol acetyltransferase activity in lung and spleen extracts from young rats that had initially received a single i.p. in utero injection of 25 μg (0.5 μg/μl) of a CMV-CAT expression plasmid complexed to 100 nmol DOTIM:cholesterol liposomes.
Figure 3: Chloramphenicol acetyltransferase activity in brain, lung, and liver extracts from groups of fetal rats after receiving a single i.p. or i.t. in utero injection of 25 μg (0.5 μg/μl) of a CMV-CAT expression plasmid complexed to 100 nmol DOTIM:cholesterol liposomes, or a single i.c. injection of 10 μg (0.5 μg/μl) of a CMV-CAT expression plasmid complexed to 40 nmol DOTIM:cholesterol liposomes.
Figure 4: PCR amplification of DNA from gonadal tissues.
Figure 5: (A) Peritoneal surface of a fetus harvested 24 h after a single i.p. injection of 25 μg of a CMV—gal expression plasmid complexed to 100 nmol DOTIM:cholesterol liposomes.
Figure 6

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Acknowledgements

This work was supported by NIH grants DK45917, DK49550, and HL53762. We thank Patrick Lewis for performing the hG-CSF assays.

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Authors and Affiliations

  1. Department of Medicine, University of California, San Francisco, San Francisco, 94143, CA

    Karin M.L. Gaensler & Robert J. Debs

  2. California Pacific Medical Center Research Institute, San Francisco, 94115, CA

    Guanhuan Tu & Robert J. Debs

  3. Department of Surgery, University of California, San Francisco, San Francisco, 94143, CA

    Steven Bruch, Gerald S. Lipshutz, Andrea Metkus & Michael Harrison

  4. Department of Comparative Medicine, University of Washington, Seattle, 98195, WA

    Denny Liggitt

  5. School of Pharmacy, University of Wisconsin-Madison, Madison, 53706, WI

    Timothy D. Heath

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  1. Karin M.L. Gaensler
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Correspondence to Karin M.L. Gaensler.

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Gaensler, K., Tu, G., Bruch, S. et al. Fetal gene transfer by transuterine injection of cationic liposome–DNA complexes. Nat Biotechnol 17, 1188–1192 (1999). https://doi.org/10.1038/70729

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