VSV-G envelope glycoprotein forms complexes with plasmid DNA and MLV retrovirus-like particles in cell-free conditions and enhances DNA transfection.
We have previously shown that vesicles containing the spike glycoprotein of the vesicular stomatitis virus (VSV-G) can associate efficiently with immature, non-infectious, envelope-deficient retrovirus-like particles assembled by packaging cells to produce infectious, pseudotyped viruses in cell-free conditions in vitro. We have also previously reported that VSV-G can enhance DNA lipofection efficiency by interacting with liposomes to form fusogenic, serum-stable liposomes with enhanced transfection properties. Here, we report that VSV-G can form a complex directly with naked plasmid DNA in the absence of a lipofection reagent and can thereby enhance the transfection efficiency of the naked plasmid vector. Sucrose gradient sedimentation analysis demonstrated that VSV-G can also associate with plasmid DNA and murine leukemia virus (MLV) gag-pol particles to form ternary complexes that co-sediment with high DNA transfecting activity. The increased transfection efficiency with VSV-G was dependent on the presence of the polycation (Polybrene) in the culture medium during transfection. Enhanced transfection was abolished by a neutralizing antibody to VSV-G. These results may be useful in the study of retrovirus assembly, in the further design of hybrid DNA-based retrovirus-like vectors, and in the full in vitro, cell-free assembly of infectious virus-like particles from component parts.
Okimoto
T
T
Program in Human Gene Therapy, Center for Molecular Genetics, UCSD School of Medicine, La Jolla, CA 92093-0692, USA.
Friedmann
T
T
Miyanohara
A
A
D000818:Animals; D002460:Cell Line; D002474:Cell-Free System; D017930:Genes, Reporter / Q000235:genetics; D009052:Leukemia Virus, Murine / Q000737:chemistry / Q000378:metabolism*; D008081:Liposomes / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins; D010957:Plasmids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000737:chemistry / Q000378:metabolism*
C028391:G protein, vesicular stomatitis virus; D008081:Liposomes; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D008156:Luciferases