Humane Genomics, New York, NY 10014, USA.
Scott-Ritchey Research Center, College of Veterinary Medicine, Auburn University, Auburn, AL 36849, USA.
Viruses. 2024 Oct 21;16(10):1641. doi: 10.3390/v16101641.
Vesicular stomatitis virus (VSV) is a prototype RNA virus that has been instrumental in advancing our understanding of viral molecular biology and has applications in vaccine development, cancer therapy, antiviral screening, and more. Current VSV genome plasmids for purchase or contract virus services provide limited options for modification, restricted to predefined cloning sites and insert locations. Improved methods and tools to engineer VSV will unlock further insights into long-standing virology questions and new opportunities for innovative therapies. Here, we report the design and construction of a full-length VSV genome. The 11,161 base pair synthetic VSV (synVSV) was assembled from four modularized DNA fragments. Following rescue and titration, phenotypic analysis showed no significant differences between natural and synthetic viruses. To demonstrate the utility of a synthetic virology platform, we then engineered VSV with a foreign glycoprotein, a common use case for studying viral entry and developing anti-virals. To show the freedom of design afforded by this platform, we then modified the genome of VSV by rearranging the gene order, switching the positions of VSV-P and VSV-M genes. This work represents a significant technical advance, providing a flexible, cost-efficient platform for the rapid construction of VSV genomes, facilitating the development of innovative therapies.
水疱性口炎病毒（VSV）是一种 RNA 病毒原型，它在推进我们对病毒分子生物学的理解方面发挥了重要作用，并在疫苗开发、癌症治疗、抗病毒筛选等方面有应用。目前可购买或通过合同病毒服务获得的 VSV 基因组质粒提供了有限的修饰选择，仅限于预定义的克隆位点和插入位置。改进的 VSV 工程方法和工具将进一步深入了解长期存在的病毒学问题，并为创新疗法提供新的机会。在这里，我们报告了全长 VSV 基因组的设计和构建。全长 11161 个碱基的合成 VSV（synVSV）由四个模块化 DNA 片段组装而成。拯救和滴定后，表型分析表明天然病毒和合成病毒之间没有显著差异。为了展示合成病毒学平台的实用性，我们随后用一种外源糖蛋白工程改造了 VSV，这是研究病毒进入和开发抗病毒药物的常见用途。为了展示该平台设计的自由度，我们随后通过重新排列基因顺序，交换 VSV-P 和 VSV-M 基因的位置，对 VSV 基因组进行了修饰。这项工作代表了一项重大的技术进步，为 VSV 基因组的快速构建提供了一个灵活、经济高效的平台，有助于开发创新疗法。
