{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,3]],"date-time":"2025-12-03T17:36:34Z","timestamp":1764783394670},"reference-count":11,"publisher":"Cold Spring Harbor Laboratory","issue":"4","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cold Spring Harb Protoc"],"published-print":{"date-parts":[[2012,4]]},"abstract":"<jats:p>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\u2013DNA 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\u2013DNA complex-mediated gene transfer resembles that of VSV-G\u2013pseudotyped 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\u2013VSV-G complexes.<\/jats:p>","DOI":"10.1101\/pdb.prot068528","type":"journal-article","created":{"date-parts":[[2012,4,2]],"date-time":"2012-04-02T14:23:58Z","timestamp":1333376638000},"page":"pdb.prot068528","source":"Crossref","is-referenced-by-count":7,"title":["Preparation of Vesicular Stomatitis Virus-G (VSV-G) Conjugate and Its Use in Gene Transfer"],"prefix":"10.1101","volume":"2012","author":[{"given":"Atsushi","family":"Miyanohara","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"246","published-online":{"date-parts":[[2012,4,2]]},"reference":[{"key":"2021111008355984000_2012.4.pdb.prot068528.1","doi-asserted-by":"crossref","first-page":"6159","DOI":"10.1128\/JVI.72.7.6159-6163.1998","article-title":"Enhanced gene transfer with fusogenic liposomes containing vesicular stomatitis virus G glycoprotein","volume":"72","year":"1998","journal-title":"J Virol"},{"key":"2021111008355984000_2012.4.pdb.prot068528.2","doi-asserted-by":"crossref","first-page":"6356","DOI":"10.1128\/JVI.72.8.6356-6361.1998","article-title":"In vitro cell-free conversion of noninfectious Moloney retrovirus particles to an infectious form by the addition of the vesicular stomatitis virus surrogate envelope G protein","volume":"72","year":"1998","journal-title":"J Virol"},{"key":"2021111008355984000_2012.4.pdb.prot068528.3","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.17.8033"},{"key":"2021111008355984000_2012.4.pdb.prot068528.4","doi-asserted-by":"crossref","first-page":"1202","DOI":"10.1128\/jvi.65.3.1202-1207.1991","article-title":"Pseudotype formation of murine leukemia virus with the G protein of vesicular stomatits virus","volume":"65","year":"1991","journal-title":"J Virol"},{"key":"2021111008355984000_2012.4.pdb.prot068528.5","doi-asserted-by":"publisher","DOI":"10.1126\/science.272.5259.263"},{"key":"2021111008355984000_2012.4.pdb.prot068528.6","doi-asserted-by":"publisher","DOI":"10.1006\/mthe.2001.0443"},{"key":"2021111008355984000_2012.4.pdb.prot068528.7","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.93.21.11395"},{"key":"2021111008355984000_2012.4.pdb.prot068528.8","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.74.22.10790-10795.2000"},{"key":"2021111008355984000_2012.4.pdb.prot068528.9","doi-asserted-by":"publisher","DOI":"10.1016\/S0091-679X(08)60600-7"},{"key":"2021111008355984000_2012.4.pdb.prot068528.10","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.20.9564"},{"key":"2021111008355984000_2012.4.pdb.prot068528.11","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1099\/0022-1317-63-1-15","article-title":"The pseudotype paradox","volume":"63","year":"1982","journal-title":"J Gen Virol"}],"container-title":["Cold Spring Harbor Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/pdb.prot068528","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,10]],"date-time":"2021-11-10T18:42:17Z","timestamp":1636569737000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.cshprotocols.org\/lookup\/doi\/10.1101\/pdb.prot068528"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,4]]},"references-count":11,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2012,4,2]]},"published-print":{"date-parts":[[2012,4]]}},"alternative-id":["10.1101\/pdb.prot068528"],"URL":"https:\/\/doi.org\/10.1101\/pdb.prot068528","relation":{},"ISSN":["1940-3402","1559-6095"],"issn-type":[{"value":"1940-3402","type":"print"},{"value":"1559-6095","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,4]]}}}