Continuous high-titer HIV-1 vector production
- 7 April 2003
- journal article
- technical report
- Published by Springer Nature in Nature Biotechnology
- Vol. 21 (5) , 569-572
- https://doi.org/10.1038/nbt815
Abstract
Human immunodeficiency virus type 1 (HIV-1)–based vectors are currently made by transient transfection, or using packaging cell lines in which expression of HIV-1 Gag and Pol proteins is induced1,2,3. Continuous vector production by cells in which HIV-1 Gag-Pol is stably expressed would allow rapid and reproducible generation of large vector batches. However, attempts to make stable HIV-1 packaging cells by transfection of plasmids encoding HIV-1 Gag-Pol have resulted in cells which secrete only low levels of p24 antigen (20–80 ng/ml)4,5,6, possibly because of the cytotoxicity of HIV-1 protease7. Infection of cells with HIV-1 can result in stable virus production8; cell clones that produce up to 1,000 ng/ml secreted p24 antigen have been described9. Here we report that expression of HIV-1 Gag-Pol by a murine leukemia virus (MLV) vector allows constitutive, long-term, high-level (up to 850 ng/ml p24) expression of HIV-1 Gag. Stable packaging cells were constructed using codon-optimized HIV-1 Gag-Pol10 and envelope proteins of gammaretroviruses; these producer cells could make up to 107 293T infectious units (i.u.)/ml (20 293T i.u./cell/day) for at least three months in culture.Keywords
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