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Mechanisms of gene transfer mediated by lipoplexes associated with targeting ligands or pH-sensitive peptides

Gene Therapy volume 6, pages 1798–1807 (1999)Cite this article

Abstract

Association of a targeting ligand such as transferrin, or an endosome disrupting peptide such as GALA, with cationic liposome–DNA complexes (‘lipoplexes’) results in a significant enhancement of transfection of several cell types (Simões S et al, Gene Therapy 1998; 5: 955–964). Although these strategies can overcome some of the barriers to gene delivery by lipoplexes, the mechanisms by which they actually enhance tranfection is not known. In studies designed to establish the targeting specificity of transferrin, we found that apo-transferrin enhances transfection to the same extent as transferrin, indicating that internalization of the lipoplexes is mostly independent of transferrin receptors. These observations were reinforced by results obtained from competitive inhibition studies either by preincubating the cells with an excess of free ligand or with various ‘receptor-blocking’ lipoplexes. Transfection of cells in the presence of drugs that interfere with the endocytotic pathway provided additional insights into the mechanisms of gene delivery by transferrin- or GALA-lipoplexes. Our results indicate that transferrin-lipoplexes deliver transgenes by endocytosis primarily via a non-receptor-mediated mechanism, and that acidification of the endosomes is partially involved in this process.

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Acknowledgements

This work was supported by JNICT, PRAXIS XXI (BD 4056/94), the University of the Pacific School of Dentistry, in part by the National Institutes of Health (AI 35231), PRAXIS/PCNA/BIO/45/96, Portugal, and grant BIO4-CT97-2191 from the European Union. We thank Dr K Konopka (University of the Pacific) for helpful discussions.

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  1. V Slepushkin

    Present address: VIRxSYS, 200 Perry Parkway, Suite 1A, Gaithersburg, MD, USA

Authors and Affiliations

  1. Department of Microbiology, School of Dentistry, University of the Pacific, San Francisco, CA, USA

    S Simões, V Slepushkin, P Pires & N Düzgüneş

  2. Laboratory of Pharmaceutical Technology, Faculty of Pharmacy, University of Coimbra, Coimbra, Portugal

    S Simões & R Gaspar

  3. Center for Neurosciences, University of Coimbra, Coimbra, Portugal

    S Simões, P Pires, R Gaspar & MC Pedroso de Lima

  4. Department of Biochemistry, University of Coimbra, Coimbra, Portugal

    P Pires & MC Pedroso de Lima

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Simões, S., Slepushkin, V., Pires, P. et al. Mechanisms of gene transfer mediated by lipoplexes associated with targeting ligands or pH-sensitive peptides. Gene Ther 6, 1798–1807 (1999). https://doi.org/10.1038/sj.gt.3301015

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