1. Inhibition of HIV-1 replication by an anti-tat hammerhead ribozyme.
- Author
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Jackson WH Jr, Moscoso H, Nechtman JF, Galileo DS, Garver FA, and Lanclos KD
- Subjects
- Animals, CD4-Positive T-Lymphocytes metabolism, COS Cells, Chloramphenicol O-Acetyltransferase genetics, Chloramphenicol O-Acetyltransferase metabolism, Genetic Vectors genetics, Moloney murine leukemia virus genetics, Plasmids genetics, RNA-Directed DNA Polymerase metabolism, Transduction, Genetic genetics, Transfection genetics, Viral Proteins antagonists & inhibitors, beta-Galactosidase genetics, tat Gene Products, Human Immunodeficiency Virus, Gene Products, tat antagonists & inhibitors, HIV-1 growth & development, RNA, Catalytic metabolism
- Abstract
Tat is a virally expressed regulatory protein involved in the replication of HIV-1, the etiological agent of AIDS. To investigate the effect of tat inhibition on HIV replication, we constructed a retroviral vector to express an anti-tat hammerhead ribozyme as part of the 3' untranslated region of beta-galactosidase transcripts. Initial testing of this vector in tat-expressing COS-7 cells reduced tat activity by 85-95% as measured by tat-dependent CAT assays. Amphotropic and HIV-pseudotyped retroviral particles generated with this vector were used in HIV challenge experiments to determine the ability of this reagent to control HIV replication. CD4(+) peripheral blood lymphocytes (PBLs) stably transduced with this vector were subsequently challenged with HIV. These cells were able to resist HIV infection for up to 20 days as measured by cell death and reverse transcriptase activity. These data yield proof of principle that a pseudotyped retroviral vector can target and deliver a protective ribozyme to CD4(+) cells., (Copyright 1998 Academic Press.)
- Published
- 1998
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