Borna disease virus (BoDV), a nonsegmented, negative-sense RNA virus, establishes persistent infection and replicates in the cell nucleus. Since BoDV genomic RNA exists as episomal RNA, the host genome is not invaded by BoDV infection. These unique features make BoDV a promising gene delivery system as an RNA virus-based episomal vector (REVec). Previously, the stable expression of genes of interest in vitro and in vivo using a REVec was reported. For the clinical application of a REVec, the fundamental properties under various physical and chemical conditions must be determined to develop purification processes, supply chains, and biosafety management. This study investigated the effects of the following conditions on the inducibility of transmission-defective ΔG-REVec: freeze–thaw cycles, dehydration, UV, temperature, pH, and reagents for virucides and laboratory experiments. Although the titer of ΔG-REVec was not influenced by the freeze–thaw process or 5 minute incubation at ≤50°C, ΔG-REVec was significantly inactivated by incubation at ≥70°C for 5 minutes. The induction titer of ΔG-REVec was decreased by long-term incubation, dehydration, and UV irradiation in a temperature- and time-dependent manner. ΔG-REVec was sensitive to lower pH and inactivated by chemical reagents under general conditions. These results provide important knowledge for developing the clinical use of REVec and biosafety management.

中文翻译：

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基于博尔纳病病毒的附加型载体在物理和化学刺激下的稳定性

博尔纳病病毒 (BoDV) 是一种非分段的负义 RNA 病毒，可建立持续感染并在细胞核中复制。由于 BoDV 基因组 RNA 作为附加体 RNA 存在，因此宿主基因组不会被 BoDV 感染侵入。这些独特的功能使 BoDV 作为基于 RNA 病毒的附加型载体 (REVec) 成为有前途的基因传递系统。以前，报道了使用 REVec 在体外和体内稳定表达感兴趣的基因。对于 REVec 的临床应用，必须确定各种物理和化学条件下的基本特性，以开发纯化工艺、供应链和生物安全管理。本研究调查了以下条件对透射缺陷型 ΔG-REVec 诱导性的影响：冻融循环、脱水、紫外线、温度、pH、以及用于杀病毒剂和实验室实验的试剂。尽管 ΔG-REVec 的滴度不受冻融过程或在 ≤50°C 下孵育 5 分钟的影响，但 ΔG-REVec 在≥70°C 下孵育 5 分钟显着失活。ΔG-REVec 的诱导效价因长期孵育、脱水和紫外线照射而以温度和时间依赖性方式降低。ΔG-REVec 对较低的 pH 值敏感，在一般条件下被化学试剂灭活。这些结果为开发 REVec 的临床应用和生物安全管理提供了重要知识。ΔG-REVec 的诱导效价因长期孵育、脱水和紫外线照射而以温度和时间依赖性方式降低。ΔG-REVec 对较低的 pH 值敏感，在一般条件下被化学试剂灭活。这些结果为开发 REVec 的临床应用和生物安全管理提供了重要知识。ΔG-REVec 的诱导效价因长期孵育、脱水和紫外线照射而以温度和时间依赖性方式降低。ΔG-REVec 对较低的 pH 值敏感，在一般条件下被化学试剂灭活。这些结果为开发 REVec 的临床应用和生物安全管理提供了重要知识。