scholarly journals Propagating and Detecting an Infectious Molecular Clone of Maedi-visna Virus that Expresses Green Fluorescent Protein

10.3791/3483 ◽  
2011 ◽  
Author(s):  
Stefán R. Jónsson ◽  
Valgerdur Andrésdóttir
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Infectious Molecular Clone ◽  
Molecular Clone ◽  
Visna Virus ◽  
Maedi Visna Virus ◽  
Green Fluorescent

Construction and characterization of an infectious molecular clone of maedi-visna virus that expresses green fluorescent protein

2010 ◽  
Vol 168 (1-2) ◽  
pp. 98-102 ◽  
Author(s):  
Hólmfrídur Sunna Gudmundsdóttir ◽  
Katrín Olafsdóttir ◽  
Sigrídur Rut Franzdóttir ◽  
Valgerdur Andrésdóttir
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Infectious Molecular Clone ◽  
Molecular Clone ◽  
Visna Virus ◽  
Maedi Visna Virus ◽  
Green Fluorescent

scholarly journals 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 ◽  
Vol 12 (9) ◽  
pp. 805
Author(s):  
Christopher Janich ◽  
Daniel Ivanusic ◽  
Julia Giselbrecht ◽  
Elena Janich ◽  
Shashank Reddy Pinnapireddy ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Plasma Membrane ◽  
Green Fluorescent Protein ◽  
Nucleic Acid ◽  
Fluorescent Protein ◽  
Cationic Lipid ◽  
Nucleic Acid Delivery ◽  
Molecular Clone ◽  
Green Fluorescent ◽  
Hiv 1

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.

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