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• W2148879761 abstract "The structural motif formed between a hammerhead ribozyme and its substrate consists of three RNA double helices In which the sequence 5′ to the XUY is termed helix I and the sequence 3′ to the XUY helix III. Two hammerhead ribozymes targeted to the tat gene of HIV-1 SF2 were designed to study target specificity and the potential effect of helix I mismatch on ribozyme efficacy both in vitro and in vivo. The first ribozyme (Rz1) targeted to the 5′ splicing region of the tat gene was designed to cleave GUC*A. In HIV-11MB the A Is changed to a G. The second ribozyme (Rz2) was targeted to the translational Initiation region of the tat gene which is highly conserved among a variety of HIV-1 isolates, Including both HIV-1 SF2 and HIV-1 IIIB. In vitro cleavage studies demonstrated that Rz1 efficiently cleaved HIV-1 SF2 substrate RNA, but not HIV-1 IIIB, presumably due to the base change from A to G. In contrast, Rz2 cleaved HIV-1 SF2 or HIV-1 IIIB substrate with equal efficiency. Both ribozymes were cloned into the 3′ untranslated region of the neomycin gene (neo) within the pSV2neo vector and transfected into the SupTl human CD4+ T cell line. Following selection, stable transfectants were challenged with either HIV-1 SF2 or HIV-1 IIIB virus. While Rz1-expressing cells were significantly protected from HIV-1 SF2 infection, they exhibited no protection when infected with HIV-1 IIIB virus. In contrast, Rz2 was effective in inhibiting the replication of both HIV-1 SF2 and HIV-1 IIIB In SupT1 cells. Expression of both ribozymes in these cells was demonstrated by Northern analysis. RT-PCR sequencing analysis confirmed the respective HIV-1 target sequence integrity. These data demonstrate the importance of the first base pair distal to the XUY within helix I of the hammerhead structure for both In vitro and in vivo ribozyme activities and imply that the effectiveness of the anti-HIV-1 ribozymes against appropriate target sequences is due to their catalytic activities rather than any antisense effect" @default.
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• W2148879761 date "1995-01-01" @default.
• W2148879761 modified "2023-09-22" @default.
• W2148879761 title "Target sequence-specific inhibition of Hiv- replication by ribozymes directed to<i>tat</i>RNA" @default.
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• W2148879761 doi "https://doi.org/10.1093/nar/23.15.2909" @default.
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