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HIV-1 RT-dependent DNAzyme expression inhibits HIV-1 replication without the emergence of escape viruses.

ABSTRACT: 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)D?ARMtR(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.

SUBMITTER: Sugiyama R

PROVIDER: S-EPMC3025543 | biostudies-literature | 2011 Jan

REPOSITORIES: biostudies-literature

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HIV-1 RT-dependent DNAzyme expression inhibits HIV-1 replication without the emergence of escape viruses.

Sugiyama Ryuichi R Hayafune Masaaki M Habu Yuichiro Y Yamamoto Norio N Takaku Hiroshi H

Nucleic acids research 20100909 2

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), ...[more]

PMID: 20833635

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