A packaging cell line for lentivirus vectors.
Lentivirus vectors can transduce dividing and nondividing cells. Using three-plasmid transient transfections, high-titer (>10(9) IU/ml) recombinant lentivirus vectors pseudotyped with vesicular stomatitis virus G (VSV-G) protein can be generated (T. Kafri et al., Nat. Genet. 17:314-317, 1997; H. Miyoshi et al., Proc. Natl. Acad. Sci. USA 94:10319-10323, 1997; L. Naldini et al., Science 272:263-267, 1996). The recombinant lentiviruses can efficiently infect brain, liver, muscle, and retinal tissue in vivo. Furthermore, the transduced tissues demonstrated long-term expression of reporter genes in immunocompetent rodents. We now report the generation of a tetracycline-inducible VSV-G pseudotyped lentivirus packaging cell line which can generate virus particles at titers greater than 10(6) IU/ml for at least 3 to 4 days. The vector produced by the inducible cell line can be concentrated to titers of 10(9) IU/ml and can efficiently transduce nondividing cells in vitro and in vivo. The availability of a lentivirus packaging cell line will significantly facilitate the production of high-titer lentivirus vectors for gene therapy and study of human immunodeficiency virus biology.
Kafri
T
T
Laboratory of Genetics, The Salk Institute, La Jolla, California 92037, USA.
van Praag
H
H
Ouyang
L
L
Gage
F H
FH
Verma
I M
IM
D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D002460:Cell Line; D005260:Female; D005822:Genetic Vectors; D016325:HIV Long Terminal Repeat; D016086:Lentivirus / Q000235:genetics / Q000502:physiology*; D008562:Membrane Glycoproteins; D051381:Rats; D011916:Rats, Inbred F344; D013752:Tetracycline / Q000494:pharmacology; D014759:Viral Envelope Proteins / Q000235:genetics*; D019065:Virus Assembly
D002087:Butyrates; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D013752:Tetracycline