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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Miyanohara, Atsushi |
| Abstract | The fusiogenic envelope G glycoprotein of the vesicular stomatitis virus (VSV-G) that has been used to pseudotype retrovirus and lentivirus vectors can be used alone as an efficient vehicle for gene transfer. VSV-G protein is secreted into the culture medium as sendimentable vesicles from cells transfected with a VSV-G expression plasmid in the absence of other viral components. The VSV-G vesicles in the conditioned medium can be partially purified by pelleting through sucrose cushion ultracentrifugation. Protein-DNA complexes are formed by mixing the VSV-G vesicles with naked plasmid DNA. Such complexes show markedly enhanced transfection efficiency when added to the culture medium of recipient cells. The cell tropism of VSV-G-DNA complex-mediated gene transfer resembles that of VSV-G-pseudotyped retrovirus and lentivirus vectors, and the complex is therefore particularly useful for transfection of cells that are refractory to other methods. Still, some cells are refractory to VSV-G-mediated transfection. It should also be noted that overdose of VSV-G can be quite toxic to the recipient cells. The primitive complexes formed by mixing a viral fusiogenic envelope protein with naked DNA may represent a step toward fusing useful features of viral and nonviral vectors for safer and more efficient gene transfer. This protocol describes simple methods for preparation of VSV-G and for gene transfer with DNA-VSV-G complexes. |
| File Format | HTM / HTML |
| ISSN | 19403402 |
| Issue Number | 4 |
| Volume Number | 2012 |
| e-ISSN | 15596095 |
| Journal | Cold Spring Harbor Protocols |
| Language | English |
| Publisher | Cold Spring Harbor Laboratory Press |
| Publisher Date | 2012-04-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Clinical Laboratory Techniques Dna-binding Proteins Isolation & Purification Metabolism Dna Membrane Glycoproteins Transfection Methods Viral Envelope Proteins Cell Line Culture Media Chemistry Genetics Humans Plasmids Protein Binding Vesiculovirus Journal Article Research Support, N.i.h., Extramural |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biochemistry, Genetics and Molecular Biology |
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