An inducible packaging cell system for safe, efficient lentiviral vector production in the absence of HIV-1 accessory proteins

被引:51
作者
Pacchia, AL
Adelson, ME
Kaul, M
Ron, Y
Dougherty, JP
机构
[1] Robert Wood Johnson Med Sch, Dept Mol Genet & Microbiol, Piscataway, NJ 08854 USA
[2] Rutgers State Univ, Grad Program Microbiol & Mol Genet, New Brunswick, NJ 08903 USA
关键词
HIV-1; lentiviral vector; cell line; regulated viral gene expression; inducible; ecdysone; GFP; gene therapy; transgene; nondividing cells; HeLa cells;
D O I
10.1006/viro.2000.0787
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Lentiviral vectors based on human immunodeficiency virus type 1 (HIV-1) possess the ability to deliver exogenous genes to both dividing and nondividing cells and to subsequently establish a stable provirus in these target cells, which can allow long-term expression of the transferred gene. Herein we describe a stable packaging cell line that is devoid of HIV-1 tat, vif, vpr, vpu, and nef. In order to avoid any risk of cytotoxicity associated with constitutive expression of HIV-I protease or the VSV-G envelope protein, transcription of the packaging and envelope constructs was tightly controlled by employing the ecdysone-inducible system. Using this cell line, we have been able to consistently generate concentrated pseudotyped vector virus stocks with titers in the range of 10(8) IU/ml, which can efficiently transduce actively dividing and growth-arrested cells in vitro. This novel packaging cell line for lentiviral vectors facilitates the production of high-titer virus stocks in the absence of replication-competent virus and provides us with an important tool for use in future gene transfer studies. (C) 2001 Academic Press.
引用
收藏
页码:77 / 86
页数:10
相关论文
共 33 条
[1]  
ADELSON ME, UNPUB
[2]   A SMALL ELEMENT FROM THE MASON-PFIZER MONKEY VIRUS GENOME MAKES HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 EXPRESSION AND REPLICATION REV-INDEPENDENT [J].
BRAY, M ;
PRASAD, S ;
DUBAY, JW ;
HUNTER, E ;
JEANG, KT ;
REKOSH, D ;
HAMMARSKJOLD, ML .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (04) :1256-1260
[3]   VESICULAR STOMATITIS-VIRUS G GLYCOPROTEIN PSEUDOTYPED RETROVIRAL VECTORS - CONCENTRATION TO VERY HIGH-TITER AND EFFICIENT GENE-TRANSFER INTO MAMMALIAN AND NONMAMMALIAN CELLS [J].
BURNS, JC ;
FRIEDMANN, T ;
DRIEVER, W ;
BURRASCANO, M ;
YEE, JK .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (17) :8033-8037
[4]   ANALYSIS OF MUTATION IN HUMAN-CELLS BY USING AN EPSTEIN-BARR-VIRUS SHUTTLE SYSTEM [J].
DUBRIDGE, RB ;
TANG, P ;
HSIA, HC ;
LEONG, PM ;
MILLER, JH ;
CALOS, MP .
MOLECULAR AND CELLULAR BIOLOGY, 1987, 7 (01) :379-387
[5]   A third-generation lentivirus vector with a conditional packaging system [J].
Dull, T ;
Zufferey, R ;
Kelly, M ;
Mandel, RJ ;
Nguyen, M ;
Trono, D ;
Naldini, L .
JOURNAL OF VIROLOGY, 1998, 72 (11) :8463-8471
[6]   Recombinant retroviruses pseudotyped with the vesicular stomatitis virus G glycoprotein mediate both stable gene transfer and pseudotransduction in human peripheral blood lymphocytes [J].
Gallardo, HF ;
Tan, C ;
Ory, D ;
Sadelain, M .
BLOOD, 1997, 90 (03) :952-957
[7]   Requirements for efficient production and transduction of human immunodeficiency virus type 1-based vectors [J].
Gasmi, M ;
Glynn, J ;
Jin, MJ ;
Jolly, DJ ;
Yee, JK ;
Chen, ST .
JOURNAL OF VIROLOGY, 1999, 73 (03) :1828-1834
[8]  
Gorman C., 1985, DNA CLONING PRACTICA, V1, P143
[9]   TIGHT CONTROL OF GENE-EXPRESSION IN MAMMALIAN-CELLS BY TETRACYCLINE-RESPONSIVE PROMOTERS [J].
GOSSEN, M ;
BUJARD, H .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (12) :5547-5551
[10]  
JONES HW, 1971, OBSTET GYNECOL, V38, P945