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Janich C, Ivanusic D, Giselbrecht J, et al. Efficient Transfection of Large Plasmids Encoding HIV-1 into Human Cells-A High Potential Transfection System Based on a Peptide Mimicking Cationic Lipid. Pharmaceutics. 2020;12(9)doi: 10.3390/pharmaceutics12090805.
Janich, C., Ivanusic, D., Giselbrecht, J., Janich, E., Pinnapireddy, S. R., Hause, G., Bakowsky, U., Langner, A., & Wölk, C. (2020). Efficient Transfection of Large Plasmids Encoding HIV-1 into Human Cells-A High Potential Transfection System Based on a Peptide Mimicking Cationic Lipid. Pharmaceutics, 12(9), . https://doi.org/10.3390/pharmaceutics12090805
Janich, Christopher, et al. "Efficient Transfection of Large Plasmids Encoding HIV-1 into Human Cells-A High Potential Transfection System Based on a Peptide Mimicking Cationic Lipid." Pharmaceutics vol. 12,9 (2020). doi: https://doi.org/10.3390/pharmaceutics12090805
Janich C, Ivanusic D, Giselbrecht J, Janich E, Pinnapireddy SR, Hause G, Bakowsky U, Langner A, Wölk C. Efficient Transfection of Large Plasmids Encoding HIV-1 into Human Cells-A High Potential Transfection System Based on a Peptide Mimicking Cationic Lipid. Pharmaceutics. 2020 Aug 25;12(9). doi: 10.3390/pharmaceutics12090805. PMID: 32854383; PMCID: PMC7559901.
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Pharmaceutics. 2020 Aug 25;12(9). doi: 10.3390/pharmaceutics12090805.

Efficient Transfection of Large Plasmids Encoding HIV-1 into Human Cells-A High Potential Transfection System Based on a Peptide Mimicking Cationic Lipid.

Pharmaceutics

Christopher Janich, Daniel Ivanusic, Julia Giselbrecht, Elena Janich, Shashank Reddy Pinnapireddy, Gerd Hause, Udo Bakowsky, Andreas Langner, Christian Wölk

Affiliations

  1. Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Strasse 4, 06120 Halle (Saale), Germany.
  2. Department of Pharmaceutics and Biopharmaceutics, University of Marburg, Robert-Koch-Str. 4, 35037 Marburg, Germany.
  3. Robert Koch Institute, Division 18: HIV and Other Retroviruses, Nordufer 20, 13353 Berlin, Germany.
  4. Biocenter, MLU Halle-Wittenberg, Weinbergweg 22, 06120 Halle (Saale), Germany.
  5. Institute of Pharmacy, Pharmaceutical Technology, Faculty of Medicine, Leipzig University, Eilenburger Strasse 15a, 04317 Leipzig, Germany.

PMID: 32854383 PMCID: PMC7559901 DOI: 10.3390/pharmaceutics12090805

Abstract

One major disadvantage of nucleic acid delivery systems is the low transfection or transduction efficiency of large-sized plasmids into cells. In this communication, we demonstrate the efficient transfection of a 15.5 kb green fluorescent protein (GFP)-fused HIV-1 molecular clone with a nucleic acid delivery system prepared from the highly potent peptide-mimicking cationic lipid OH4 in a mixture with the phospholipid DOPE (co-lipid). For the transfection, liposomes were loaded using a large-sized plasmid (15.5 kb), which encodes a replication-competent HIV type 1 molecular clone that carries a Gag-internal green fluorescent protein (HIV-1 JR-FL Gag-iGFP). The particle size and charge of the generated nanocarriers with 15.5 kb were compared to those of a standardized 4.7 kb plasmid formulation. Stable, small-sized lipoplexes could be generated independently of the length of the used DNA. The transfer of fluorescently labeled pDNA-HIV1-Gag-iGFP in HEK293T cells was monitored using confocal laser scanning microscopy (cLSM). After efficient plasmid delivery, virus particles were detectable as budding structures on the plasma membrane. Moreover, we observed a randomized distribution of fluorescently labeled lipids over the plasma membrane. Obviously, a significant exchange of lipids between the drug delivery system and the cellular membranes occurs, which hints toward a fusion process. The mechanism of membrane fusion for the internalization of lipid-based drug delivery systems into cells is still a frequently discussed topic.

Keywords: HIV; cationic lipids; gene therapy; large plasmids; membrane fusion; transfection

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