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

Virology. 2001 Mar 30;282(1):77-86. doi: 10.1006/viro.2000.0787.

Abstract

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-1 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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Line*
  • Ecdysone / pharmacology
  • Gene Products, tat / deficiency
  • Gene Products, tat / genetics
  • Gene Products, vif / deficiency
  • Gene Products, vif / genetics
  • Gene Products, vpr / deficiency
  • Gene Products, vpr / genetics
  • Genetic Vectors*
  • Green Fluorescent Proteins
  • HIV-1 / genetics*
  • Human Immunodeficiency Virus Proteins
  • Humans
  • Lentivirus / genetics*
  • Luminescent Proteins / genetics
  • Membrane Glycoproteins*
  • Transduction, Genetic
  • Transfection
  • Viral Envelope Proteins / genetics
  • Viral Proteins / genetics
  • Viral Regulatory and Accessory Proteins / deficiency
  • Viral Regulatory and Accessory Proteins / genetics
  • Virus Replication
  • tat Gene Products, Human Immunodeficiency Virus
  • vif Gene Products, Human Immunodeficiency Virus
  • vpr Gene Products, Human Immunodeficiency Virus

Substances

  • G protein, vesicular stomatitis virus
  • Gene Products, tat
  • Gene Products, vif
  • Gene Products, vpr
  • Human Immunodeficiency Virus Proteins
  • Luminescent Proteins
  • Membrane Glycoproteins
  • Viral Envelope Proteins
  • Viral Proteins
  • Viral Regulatory and Accessory Proteins
  • tat Gene Products, Human Immunodeficiency Virus
  • vif Gene Products, Human Immunodeficiency Virus
  • vpr Gene Products, Human Immunodeficiency Virus
  • vpu protein, Human immunodeficiency virus 1
  • Green Fluorescent Proteins
  • Ecdysone