Intracellular immunization against SIVmac utilizing a hairpin ribozyme.
A hairpin ribozyme targeting the 3' LTR region (9456) of SIVmac238 was cloned into a murine retroviral vector. This target sequence is conserved among various SIV, as well as most HIV-2, strains. The ribozyme cassette is driven from a polymerase III promoter, that of the human tRNAval gene. Hybrid human B-/T-cell lines (CEM/174) were transduced with the retroviral constructs and selected for G418 resistance. Cells stably expressing the 9456 ribozyme exhibited long-term resistance to infection by a pathogenic molecular clone of SIV and two strains of HIV-2. The ribozyme was also able to effectively reduce the proviral DNA burden. Its efficient protection against SIV/HIV-2 infection constitutes an important step toward evaluating ribozyme gene therapy in a primate model.
Heusch
M
M
Department of Biology, University of California at San Diego, La Jolla 92093-0665, USA.
Kraus
G
G
Johnson
P
P
Wong-Staal
F
F
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004279:DNA, Viral; D015316:Genetic Therapy; D015498:HIV-2 / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015302:Simian Immunodeficiency Virus / Q000378:metabolism*
D004279:DNA, Viral; D016337:RNA, Catalytic; D011994:Recombinant Proteins