RNA cleavage by a mammalian tRNA 3'processing endoribonuclease (3'tRNase) reduces HIV-1 expression
Open Access
- 1 November 2002
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Symposium Series
- Vol. 2 (1) , 297-298
- https://doi.org/10.1093/nass/2.1.297
Abstract
We examined the suppression of HIV-1 expression by cleavage of the HIV-1 RNA gene, using a mammalian tRNA 3′processing endoribonuclease (3′tRNase) as an External Guide Sequence oligozyme (EGS) in vivo. The enzyme can also recognize and cleave a target RNA that forms a pre-tRNA like complex with a substrate RNA. We designed an EGS to specifically recognize the packaging signal region (p.s.) and the upstream portion of the gag region (g.s.) of HIV-1 mRNA, and constructed EGS expression vectors that used the tRNAmet promoter as an expression cassette for the EGS. Their cleavage activities were assessed in cell culture, using a transient assay system. The EGS efficiencies were determined by co-transfection of the EGS expression vectors and the HTV-1 gene plasmid vector (pNL-luc) into COS cells. The assay results showed a significant inhibition of the HIV-1 gag p24 antigen expression. In addition, we constructed MLV-based VSV-G pseudotyped retrovirus vectors that express the EGS. They were co-infected into MT-4 cells with the HIV-1 virus (NL4-3). These EGS expression retrovirus vectors exhibited a similar inhibitory effectKeywords
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