Effective inhibition of HIV-1 replication in cultured cells by external guide sequences and ribonuclease.
We examined the suppression effect of HIV-1 expression by cleavage of the HIV-1 RNA gene, using the catalytic RNA subunit RNase P and the 3'-half tRNA [External Guide Sequence (EGS)] in vivo. The vectors were designed to express an anti-HIV EGS, U5, which targets the 5' leader sequence. We constructed an EGS expression vector, that used the tRNA(met) or U6 promoter as an expression cassette for EGS. RNase P cleaves the targeted HIV-1 mRNA when they are in a complex with the EGS. To test the antiviral efficacy of these EGS vectors, we have cotransfected into COS cells with the HIV-1 proviral DNA (pNL4-3 Luc) and the plasmid expressing the EGS from the tRNA(met) or U6 promoter. HIV-1 expression was inhibited by the tRNA-EGS-1 and U6-EGS-1 from the tRNA(met) and U6 promoters, respectively. No difference in the inhibitory effects on HIV-1 expression between the tRNA(met) and U6 promoters could be detected.
Endo
Y
Y
Department of Industrial Chemistry, Faculty of Engineering, Chiba Institute of Technology, 2-17-1 Tsudanuma, Narashino, Chiba 275-0016, Japan.
Miyano-Kurosaki
N
N
Kitano
M
M
Habu
Y
Y
Takaku
H
H
D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D004722:Endoribonucleases / Q000378:metabolism*; D015870:Gene Expression; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D046911:Macromolecular Substances; D011401:Promoter Regions, Genetic; D016337:RNA, Catalytic / Q000378:metabolism*; D012343:RNA, Transfer / Q000737:chemistry*; D012358:RNA, Transfer, Met / Q000235:genetics; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism; D043262:Ribonuclease P; D014779:Virus Replication
D046911:Macromolecular Substances; D016337:RNA, Catalytic; D012358:RNA, Transfer, Met; D012367:RNA, Viral; D012343:RNA, Transfer; D004722:Endoribonucleases; D043262:Ribonuclease P