Efficient gene transfer by a human immunodeficiency virus type 1 (HIV-1)-derived vector utilizing a stable HIV packaging cell line.
By transfecting fibroblast cells with an HIV-1-MN molecular clone with a deletion of the major packaging sequence, we have developed a stable HIV-1 packaging cell line, psi 422, psi 422 cells form syncytia with CD4-positive cells, correctly express HIV-1 structural proteins, and produce a large amount of mature particles with normal reverse transcriptase activity. Yet these particles, in which RNA was not detected by reverse transcriptase-PCR, are not infectious. When stably transfected with an HIV-1-based retroviral vector, the psi 422 cell line produces virions capable of transducing CD4-positive cells with high efficiency (up to 10(5) cells per ml). The availability of this stable noninfectious HIV-1 packaging cell line capable of generating high-titer HIV-1 vectors represents a new step toward the use of an HIV-1 delivery system in gene therapy.
Corbeau
P
P
Department of Medicine and Biology, University of California at San Diego, La Jolla 92093-0665, USA.
Kraus
G
G
Wong-Staal
F
F
D001483:Base Sequence; D015704:CD4 Antigens / Q000502:physiology; D002460:Cell Line; D017931:DNA Primers; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D015726:Giant Cells; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D016133:Polymerase Chain Reaction; D014162:Transfection / Q000379:methods*; D014764:Viral Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D015678:Viral Structural Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D014771:Virion / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology
D015704:CD4 Antigens; D017931:DNA Primers; D014764:Viral Proteins; D015678:Viral Structural Proteins