腺病毒载体缺失病毒相关 RNA 表达可增强 shRNA 活性，抑制丙型肝炎病毒复制。
Adenovirus vectors lacking virus-associated RNA expression enhance shRNA activity to suppress hepatitis C virus replication.
机构信息
Laboratory of Molecular Genetics, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan.
Department of Infectious Diseases, Hamamatsu University School of Medicine, Hamamatsu 431-3192, Japan.
出版信息
Sci Rep. 2013 Dec 20;3:3575. doi: 10.1038/srep03575.
First-generation adenovirus vectors (FG AdVs) expressing short-hairpin RNA (shRNA) effectively downregulate the expressions of target genes. However, this vector, in fact, expresses not only the transgene product, but also virus-associated RNAs (VA RNAs) that disturb cellular RNAi machinery. We have established a production method for VA-deleted AdVs lacking expression of VA RNAs. Here, we showed that the highest shRNA activity was obtained when the shRNA was inserted not at the popularly used E1 site, but at the E4 site. We then compared the activities of shRNAs against hepatitis C virus (HCV) expressed from VA-deleted AdVs or conventional AdVs. The VA-deleted AdVs inhibited HCV production much more efficiently. Therefore, VA-deleted AdVs were more effective than the currently used AdVs for shRNA downregulation, probably because of the lack of competition between VA RNAs and the shRNAs. These VA-deleted AdVs might enable more effective gene therapies for chronic hepatitis C.
第一代腺病毒载体（FGAdV）表达短发夹 RNA（shRNA）可有效下调靶基因的表达。然而，该载体实际上不仅表达转基因产物，还表达干扰细胞 RNAi 机制的病毒相关 RNA（VA RNA）。我们已经建立了一种生产方法，用于生产缺乏 VA RNA 表达的 VA 缺失型 AdV。在这里，我们表明，当 shRNA 不是插入到常用的 E1 位点，而是插入到 E4 位点时，可获得最高的 shRNA 活性。然后，我们比较了 VA 缺失型 AdV 或常规 AdV 表达的针对丙型肝炎病毒（HCV）的 shRNA 的活性。VA 缺失型 AdV 更有效地抑制 HCV 的产生。因此，VA 缺失型 AdV 比目前使用的 AdV 更有效地用于 shRNA 下调，可能是因为 VA RNA 与 shRNA 之间不存在竞争。这些 VA 缺失型 AdV 可能为慢性丙型肝炎的更有效的基因治疗提供了可能。
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