修饰DNA酶对流感病毒RNA (PB2 mRNA)表达的抑制作用

H Takahashi, T Abe, K Takai, H Takaku
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引用次数: 3

摘要

DNA酶是切割rna的单链DNA分子。Santro等人于1997年测定了一种DNA酶的结构和催化结构域。在这项研究中,我们设计了几种针对甲型流感病毒PB2 mRNA翻译起始区的DNA酶(PB2Dz),并检测了它们的裂解动力学、核酸酶抗性和荧光素酶基因报告试验。使用合成底物,这些DNA酶被证明具有裂解活性,这取决于底物识别结构域的长度。为了赋予血清核酸酶对DNA酶的抗性,我们设计了一种新型DNA酶,在每个末端都有N3'-P5'磷酸化修饰(PB2Dz-N)。我们在体内检测了这种DNA酶的活性。本研究中使用的DNA酶抑制COS细胞中pb2荧光素酶基因的表达。这些结果表明,DNA酶作为甲型流感病毒的基因灭活剂可能有用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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.

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