Inhibition of influenza virus RNA (PB2 mRNA) expression by a modified DNA enzyme.
DNA enzymes are RNA-cleaving single stranded DNA molecules. The structure and the catalytic domain of a DNA enzyme were determined by Santro et al. in 1997. In this study, we have designed several types of DNA enzymes (PB2Dz) targeted to the PB2 mRNA translation initiation region of influenza A virus, and examined their cleavage kinetics, nuclease resistance, and a luciferase gene reporter assay. Using a synthetic substrate, these DNA enzymes were shown to have cleavage activity that is dependent on the length of the substrate recognition domain. To confer serum nuclease resistance to the DNA enzymes, we designed a new type of DNA enzyme that has the N3'-P5' phosphoramidate modification (PB2Dz-N) at each terminal. We examined the activity of this DNA enzyme in vivo. The DNA enzymes used in this study inhibited the expression of the PB2-luciferase gene in COS cells. These results suggest that DNA enzymes are potentially useful as gene inactivating agents of influenza A virus.
Takahashi
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H
Department of Industrial Chemistry, Chiba Institute of Technology, 2-17-1 Tsudanuma, Narashino, Chiba 275-0016, Japan.
Abe
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Takai
K
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Takaku
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H
D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D021881:DNA, Catalytic; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D066298:In Vitro Techniques; D009980:Influenza A virus / Q000235:genetics* / Q000378:metabolism*; D007700:Kinetics; D008156:Luciferases / Q000235:genetics; D009690:Nucleic Acid Conformation; D012333:RNA, Messenger / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D012324:RNA-Dependent RNA Polymerase; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics
D021881:DNA, Catalytic; D004277:DNA, Single-Stranded; C057027:PB2 protein, Influenzavirus A; D012333:RNA, Messenger; D012367:RNA, Viral; C402582:RNA-cleaving DNA 10-23; D014764:Viral Proteins; D008156:Luciferases; D012324:RNA-Dependent RNA Polymerase