Functional characterization of a U5 ribozyme: intracellular suppression of human immunodeficiency virus type 1 expression.
We have designed a ribozyme that cleaves human immunodeficiency virus type 1 (HIV-1) RNA in U5 (at nucleotide +115). This ribozyme was tested in vitro and was found to give efficient and specific digestion of RNA containing the HIV-1 U5 sequence. When the U5 ribozyme was placed into the HIV-1 genome, virus replication was suppressed in tissue culture. Introduction of this ribozyme into cells by using an amphotropic retrovirus vector significantly reduced expression of U5-containing RNA in cells chronically infected with HIV-1. Naive T cells were cocultivated with packaging cells that produce defective amphotropic retroviruses containing the U5 ribozyme. These lymphocytes were found to be partially protected from HIV-1 infection.
Dropulić
B
B
Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Lin
N H
NH
Martin
M A
MA
Jeang
K T
KT
D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D015967:Gene Expression Regulation, Viral; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016337:RNA, Catalytic / Q000235:genetics*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics*; D013379:Substrate Specificity; D014779:Virus Replication
D004274:DNA, Recombinant; D009838:Oligodeoxyribonucleotides; D016337:RNA, Catalytic; D012367:RNA, Viral