Inhibition of HIV-1 Replication by an HIV-1 Dependent Ribozyme Expression Vector with the Cre/loxP (ON/OFF) System

• Published in: Antiviral chemistry & chemotherapy Vol. 13; no. 5; pp. 273 - 281
• Main Authors: Habu, Yuichiro, Miyano-Kurosaki, Naoko, Nagawa, Takashi, Matsumoto, Norihiko, Takeuchi, Hiroaki, Takaku, Hiroshi
• Format: Journal Article
• Language: English
• Published: London, England SAGE Publications 01.10.2002; International Medical Press; Sage Publications Ltd
• Subjects: Antibiotics. Antiinfectious agents. Antiparasitic agents; Antiviral agents; Attachment Sites, Microbiological - genetics; Biological and medical sciences; CD4 antigen; Cre recombinase; Gene expression; Gene Expression Regulation; Gene Expression Regulation, Viral; Gene therapy; Genetic Engineering; Genetic Therapy; Genetic Vectors - genetics; HeLa Cells; HIV; HIV-1 - genetics; HIV-1 - physiology; Homologous recombination; Human immunodeficiency virus; Human immunodeficiency virus 1; Humans; Integrases - genetics; Integrases - metabolism; Long terminal repeat; Medical sciences; Mutagenesis, Site-Directed; Pharmacology. Drug treatments; Promoter Regions, Genetic - genetics; Recombination, Genetic - genetics; Replication; Ribonucleic acid; RNA; RNA, Catalytic - genetics; RNA, Catalytic - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; RNA, Transfer, Met - genetics; RNA, Viral - genetics; RNA, Viral - metabolism; Tat protein; Viral Proteins - genetics; Viral Proteins - metabolism; Virus Replication; HIV-1; catalytic RNA; tat; virus replication; transfection; hammerhead ribozyme; gene therapy; antiviral agent; LTR; Cre recombinase; HIV-1 virus; Homologous recombination; Promoter; Transfection; Established cell line; Antiviral; LTR sequence; Vector; Ribozyme; Retroviridae; Gene expression; Lentivirus; Biological activity; Virus; Replication; Human immunodeficiency virus; Gene therapy
• ISSN: 2040-2066; 0956-3202; 2040-2066
• DOI: 10.1177/095632020201300502

Antiviral strategies to inhibit HIV-1 replication have included the generation of gene products that provide the intracellular inhibition of an essential viral protein or RNA. When used in conjunction with the HIV-1 long terminal repeat (LTR), an inducible promoter dependent on the virus-encoded trans-activator (tat), relatively high background activity is still observed in the absence of tat (Caruso & Klatzmann, 1992; Dinges et al., 1995). In order to circumvent this problem, we used the Cre/loxP (ON/OFF) recombination system as a tool for our investigation. In the present study, we constructed a loxP-cassette vector with the ribozyme (Rz) expression portion under the control of the tRNAiMet promoter between two loxP sequences (plox-Rz-U5). We also constructed an HIV-1 LTR promoter-driven Cre recombinase gene (pLTR-Cre). These vectors were triple-transfected into HeLa CD4 cells with the HIV-1 pseudo-type viral expression vector. Basal activity was not detectable before HIV-1 infection. The LTR-dependent Cre protein product in HIV-1 infected HeLa CD4 cells expressed the ribozyme by inducing loxP homologous recombination, which strongly inhibited the HIV-1 gene expression. These results demonstrate the potential of an anti-ribozyme with the Cre/loxP system for controlling HIV-1 infection via gene therapy.