A new modified DNA enzyme that targets influenza virus A mRNA inhibits viral infection in cultured cells

FEBS Letters

Hayashi, Mieko¹  Takahashi, Hitoshi¹  Abe, Takayuki¹  Takaku, Hiroshi¹  Habu, Yuichiro¹  Hamazaki, Hiroyuki¹  Miyano-Kurosaki, Naoko¹ 

[1] Department of Life and Environmental Sciences, Chiba Institute of Technology, 2-17-1 Tsudanuma, Narashino, Chiba 275-0016, Japan

关键词: Modified DNA enzyme;    N3′-P5′ phosphoramidate oligonucleotide;    Influenza A virus;    Kinetic analysis;    Nuclease resistance;    Plaque formation assay;    NP;    nucleoprotein;    PB;    basic polymerase;    Dz;    DNA enzyme;    N;    oligonucleotide N3′-P5′ phosphoramidate;    M;    2′-O-methyl-nucleoside;    I;    inactive DNA enzyme;    PR/8;    influenza virus A/Puerto Rico/8/34 (H1N1);    DOTAP;    {N-[1-(2;    3-dioleoyloxy)propyl]-N;    N;    N-trimethylammonium methylsulfate;    MDCK cell;    Madin–Darby canine kidney cell;    moi;    multiplicity of infection;    FITC;    fluorescein isothiocyanate;   

DOI  :  10.1016/S0014-5793(04)00073-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

DNA enzymes are RNA-cleaving single-stranded DNA molecules. We designed DNA enzymes targeting the PB2 mRNA translation initiation (AUG) region of the influenza A virus (A/PR/8/34). The modified DNA enzymes have one or two N3′-P5′ phosphoramidate bonds at both the 3′- and 5′-termini of the oligonucleotides, which significantly enhanced their nuclease resistance. These modified DNA enzymes had the same cleavage activity as the unmodified DNA enzymes, determined by kinetic analyses, and reduced influenza A virus replication by more than 99%, determined by plaque formation. These DNA enzymes are highly specific; their protective effect was not observed in influenza B virus (B/Ibaraki)-infected Madin–Darby canine kidney cells.

【 授权许可】

Unknown