Rossi J, Chang P, Stephens D, Ladne P, Cantin E, Zaia JA; International Conference on AIDS.

Int Conf AIDS. 1989 Jun 4-9; 5: 536 (abstract no. Th.C.O.22).

Beckman Research Institute of the City of Hope, Duarte, CA, USA

OBJECTIVE: To design and test autocatalytic RNA specific for HIV-1. METHODS: We have utilized two classes of catalytic ribozymes targeted to HIV-1 RNA: one class (RBZ) is targeted to a site within the gag gene containing part of the required autocleavage site (GAAAC----GUX). The remainder of the required sequence (CUGAXGA) is supplied in trans along with flanking sequences to promote specific hybrid formation. The second class of ribozyme (GUX-RBZ) is patterned from Haseloff and Gerlach (Nature 34: 585, 1988) and contains all required sequences for auto-cleavage (except the target site GUX) along with appropriate flanking sequences to promote hybridization to a specific GUX site in the HIV-1 RNA. RBZ and GUX-RBZ were synthesized by in vitro transcription, purified by PAGE, incubated under varying conditions with the target HIV-1 RNA transcripts, and analyzed by PAGE and autoradiography. RESULTS: GUX-RBZ targeted to gag cleaved the HIV-1 RNA more efficiently than did RBZ. The GUX-RBZ cleavage reaction is entirely magnesium dependent, optimal at 20mM MgCl2 and 45 degrees C. GUX-RBZ is catalytically active from 22 degrees C to 65 degrees C and completely cleaves large molar excesses of substrate. CONCLUSION: RBZs can be designed which can potentially target any GUX site in the HIV-1 genome and deserve evaluation as potential antiviral agents.

Publication Types:

• Meeting Abstracts

Keywords:

• Base Sequence
• DNA Primers
• Genes, gag
• HIV-1
• In Vitro
• RNA
• RNA, Catalytic
• Transcription, Genetic
• genetics
• physiology

Other ID:

• 00277789

UI: 102178679

From Meeting Abstracts