Short peptide nucleic acids (PNA) inhibit hepatitis C virus internal ribosome entry site (IRES) dependent translation in vitro
Short peptide nucleic acids (PNA) inhibit hepatitis C virus internal ribosome entry site (IRES) dependent translation in vitro
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DOI：
10.1016/j.antiviral.2008.06.011
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发表时间：
2008-12-01
影响因子：
7.6
通讯作者：
Hantz, Olivier
中科院分区：
文献类型：
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作者：
Alotte, Christine;Martin, Amaury;Hantz, Olivier
丙型肝炎病毒的内部核糖体进入位点(IRES)控制着病毒RNA合成蛋白质的起始，是反义方法的理想靶点。使用原始的双顺反子质粒，我们首先确定了从6名患者克隆的丙型肝炎病毒IRESS的序列和翻译活性是保守的，无论是否对联合治疗有应答。然后，我们验证了反义分子，即短肽核酸(PNA)，可以通过与IRES的高度保守的IIID或IV环区结合来抑制丙型肝炎病毒翻译的假设。设计了5个6-10 MET PNA。在兔网织红细胞裂解物实验中，它们强烈抑制丙型肝炎病毒IRES驱动的翻译。这种抑制具有高度的特异性，因为只有一个错配的对应PNA是无效的。相同序列的短硫代寡核苷酸不能抑制丙型肝炎病毒的翻译。当PNA分子与全长的丙型肝炎病毒基因组构建物电穿孔时，在Huh-7.5细胞中显示了抗丙型肝炎病毒的活性。以寡核苷酸为载体，将PNA导入携带丙型肝炎病毒复制子的细胞和JFH1感染的Huh-7.5细胞。(C)2008爱思唯尔B.V.保留所有权利。
The internal ribosome entry site (IRES) of hepatitis C virus (HCV) which governs the initiation of protein synthesis from viral RNA represents an ideal target for antisense approaches. Using an original bicistronic plasmid, we first established that sequence and translational activity of HCV IRESs cloned from six patients, whether responders or not to combination therapy, were conserved. We then tested the hypothesis that antisense molecules, i.e. short peptide nucleic acids (PNA), could inhibit HCV translation by binding to the highly conserved IIId or IV loop regions of the IRES. Five 6-10 met PNAs were designed. They strongly inhibit HCV IRES-driven translation in a rabbit reticulocyte lysate assay. This inhibition was highly specific since corresponding PNAs with only one mismatch were inactive. Short phosphorothioate oligonucleotides of same sequence were unable to inhibit HCV translation. PNA molecule was shown to have anti-HCV activity in Huh-7.5 cells when electroporated with a full-length HCV genome construct. Using oligonucleotide as carrier, PNA was also transfected in HCV replicon-harboring cells and in JFH1 infected Huh-7.5 cells. (C) 2008 Elsevier B.V. All rights reserved.