HIV-1 RT-dependent DNAzyme expression inhibits HIV-1 replication without the emergence of escape viruses.
DNAzymes are easier to prepare and less sensitive to chemical and enzymatic degradation than ribozymes; however, a DNA enzyme expression system has not yet been developed. In this study, we exploited the mechanism of HIV-1 reverse transcription (RT) in a DNA enzyme expression system. We constructed HIV-1 RT-dependent lentiviral DNAzyme expression vectors including the HIV-1 primer binding site, the DNA enzyme, and either a native tRNA (Lys-3), tR(M)DtR(L), or one of two truncated tRNAs (Lys-3), tR(M)DΔARMtR(L) or tR(M)D3'-endtR(L). Lentiviral vector-mediated DNAzyme expression showed high levels of inhibition of HIV-1 replication in SupT1 cells. We also demonstrated the usefulness of this approach in a long-term assay, in which we found that the DNAzymes prevented escape from inhibition of HIV. These results suggest that HIV-1 RT-dependent lentiviral vector-derived DNAzymes prevent the emergence of escape mutations.
Sugiyama
Ryuichi
R
Department of Life and Environmental Science, Chiba Institute of Technology, Narashino-shi, Chiba, Japan.
Hayafune
Masaaki
M
Habu
Yuichiro
Y
Yamamoto
Norio
N
Takaku
Hiroshi
H
D000998:Antiviral Agents / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D021881:DNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D009154:Mutation; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D012343:RNA, Transfer / Q000235:genetics; D012367:RNA, Viral / Q000378:metabolism; D048348:Reverse Transcription; D014779:Virus Replication; D054299:env Gene Products, Human Immunodeficiency Virus / Q000235:genetics
D000998:Antiviral Agents; D021881:DNA, Catalytic; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral; D054299:env Gene Products, Human Immunodeficiency Virus; D012343:RNA, Transfer