Institute of Virology, Medical University of Innsbruck, Innsbruck, Austria.
Christian Doppler Laboratory for Viral Immunotherapy of Cancer, Medical University of Innsbruck, Innsbruck, Austria.
Nat Commun. 2021 Mar 1;12(1):1362. doi: 10.1038/s41467-021-21630-5.
Therapeutic application of RNA viruses as oncolytic agents or gene vectors requires a tight control of virus activity if toxicity is a concern. Here we present a regulator switch for RNA viruses using a conditional protease approach, in which the function of at least one viral protein essential for transcription and replication is linked to autocatalytical, exogenous human immunodeficiency virus (HIV) protease activity. Virus activity can be en- or disabled by various HIV protease inhibitors. Incorporating the HIV protease dimer in the genome of vesicular stomatitis virus (VSV) into the open reading frame of either the P- or L-protein resulted in an ON switch. Here, virus activity depends on co-application of protease inhibitor in a dose-dependent manner. Conversely, an N-terminal VSV polymerase tag with the HIV protease dimer constitutes an OFF switch, as application of protease inhibitor stops virus activity. This technology may also be applicable to other potentially therapeutic RNA viruses.
如果毒性是一个问题，那么 RNA 病毒作为溶瘤剂或基因载体的治疗应用需要严格控制病毒活性。在这里，我们使用条件蛋白酶方法为 RNA 病毒提供了一种调控开关，其中至少一种对转录和复制至关重要的病毒蛋白的功能与自身催化的外源性人类免疫缺陷病毒 (HIV) 蛋白酶活性相关联。病毒活性可以通过各种 HIV 蛋白酶抑制剂来开启或关闭。将 HIV 蛋白酶二聚体整合到水疱性口炎病毒 (VSV) 的基因组中，使其位于 P 蛋白或 L 蛋白的开放阅读框中，形成了 ON 开关。在这里，病毒活性取决于蛋白酶抑制剂的共同应用，且呈剂量依赖性。相反，带有 HIV 蛋白酶二聚体的 N 端 VSV 聚合酶标签构成了 OFF 开关，因为蛋白酶抑制剂的应用会停止病毒活性。这项技术也可能适用于其他潜在的治疗性 RNA 病毒。
