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Janich, Christopher [Verfasser:in]; Ivanusic, Daniel [Verfasser:in]; Giselbrecht, Julia [Verfasser:in]; Janich, Elena [Verfasser:in]; Pinnapireddy, Shashank Reddy [Verfasser:in]; Hause, Gerd [Verfasser:in]; Bakowsky, Udo [Verfasser:in]; Langner, Andreas [Verfasser:in]; Wölk, Christian [Verfasser:in]

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

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  • Medientyp: E-Artikel
  • Titel: Efficient Transfection of Large Plasmids Encoding HIV-1 into Human Cells—A High Potential Transfection System Based on a Peptide Mimicking Cationic Lipid
  • Beteiligte: Janich, Christopher [Verfasser:in]; Ivanusic, Daniel [Verfasser:in]; Giselbrecht, Julia [Verfasser:in]; Janich, Elena [Verfasser:in]; Pinnapireddy, Shashank Reddy [Verfasser:in]; Hause, Gerd [Verfasser:in]; Bakowsky, Udo [Verfasser:in]; Langner, Andreas [Verfasser:in]; Wölk, Christian [Verfasser:in]
  • Erschienen: Basel: MDPI, [2023]
  • Erschienen in: Pharmaceutics ; 12,9, (2020)
  • Sprache: Englisch
  • Schlagwörter: membrane fusion ; transfection ; HIV ; gene therapy ; cationic lipids ; large plasmids
  • Entstehung:
  • Anmerkungen:
  • Beschreibung: 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.
  • Zugangsstatus: Freier Zugang
  • Rechte-/Nutzungshinweise: Namensnennung (CC BY)