RNA polymerase I-driven minigenome system for Ebola viruses
RNA polymerase I-driven minigenome system for Ebola viruses
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DOI：
10.1128/jvi.79.7.4425-4433.2005
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发表时间：
2005-04-01
影响因子：
5.4
通讯作者：
Flick, R
中科院分区：
文献类型：
--
作者：
Groseth, A;Feldmann, H;Flick, R
一般来说，埃博拉病毒以其在人类和非人类灵长类动物中引起严重出血热的能力而闻名。然而，尽管与丝状病毒科的其他成员具有实质性的序列同源性，但雷斯顿埃博拉病毒对非人灵长类动物显示出降低的致病性，并且从未被证明在人类中引起临床疾病，尽管其能够引起感染。为了开发一种工具来探索转录和复制在降低莱斯顿埃博拉病毒的致病性中的潜在作用，我们开发了RNA聚合酶I（Pol I）驱动的微型基因组系统。在这里，我们展示了成功的莱斯顿埃博拉病毒微型基因组拯救，包括重组，转录和复制，以及包装成后代颗粒的微型基因组转录本。Pol I驱动的莱斯顿埃博拉病毒微型基因组系统提供了比同时开发的可比较的T7驱动的莱斯顿埃博拉病毒微型基因组系统更高的信号强度和更少的背景（更高的信噪比）。成功的莱斯顿埃博拉病毒微型基因组拯救也是通过使用来自密切相关的扎伊尔埃博拉病毒或更远相关的维多利亚马尔堡湖病毒的辅助质粒实现的。在雷斯顿埃博拉病毒微型基因组系统中使用异源辅助质粒产生的报告基因表达水平远远超过同源辅助质粒产生的水平。来自不同丝状病毒种属的微小基因组和辅助质粒之间的这种比较表明，致病性和非致病性丝状病毒的核糖核蛋白复合物的转录和/或复制能力可能存在固有差异，因为这些观察结果在维多利亚湖马尔堡病毒微小基因组系统中得到证实。
In general, Ebola viruses are well known for their ability to cause severe hemorrhagic fever in both human and nonhuman primates. However, despite substantial sequence homology to other members of the family Filoviridae, Reston ebolavirus displays reduced pathogenicity for nonhuman primates and has never been demonstrated to cause clinical disease in humans, despite its ability to cause infection. In order to develop a tool to explore potential roles for transcription and replication in the reduced pathogenicity of Reston ebolavirus, we developed an RNA polymerase I (Pol I)-driven minigenome system. Here we demonstrate successful Reston ebolavirus minigenome rescue, including encapsidation, transcription, and replication, as well as the packaging of minigenome transcripts into progeny particles. The Pol I-driven Reston ebolavirus minigenome system provides a higher signal intensity with less background (higher signal-to-noise ratio) than a comparable T7-driven Reston ebolavirus minigenome system which was developed simultaneously. Successful Reston ebolavirus minigenome rescue was also achieved by the use of helper plasmids derived from the closely related Zaire ebolavirus or the more distantly related Lake Victoria marburgvirus. The use of heterologous helper plasmids in the Reston ebolavirus minigenome system yielded levels of reporter expression which far exceeded the level produced by the homologous helper plasmids. This comparison between minigenomes and helper plasmids from different filovirus species and genera indicates that inherent differences in the transcription and/or replication capacities of the ribonucleoprotein complexes of pathogenic and apathogenic filoviruses may exist, as these observations were confirmed in a Lake Victoria marburgvirus minigenome system.