Alphavirus-based expression vectors: Strategies and applications
Alphavirus-based expression vectors: Strategies and applications
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DOI：
10.1073/pnas.93.21.11371
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发表时间：
1996-10-15
影响因子：
11.1
通讯作者：
Rice, CM
中科院分区：
文献类型：
--
作者：
Frolov, I;Hoffman, TA;Rice, CM
甲病毒是一种正链RNA病毒，可在昆虫和脊椎动物细胞中介导有效的胞质基因表达。通过重组DNA技术，甲病毒RNA复制机制已被工程化用于高水平表达异源RNA和蛋白质。可复制甲病毒RNA（复制子）的扩增可以通过RNA或DNA转染来启动，并且已经开发了多种包装系统用于产生高滴度的感染性病毒颗粒。尽管通常对脊椎动物细胞具有杀细胞作用，但具有允许非致细胞病变复制的适应性突变的变体已从持续感染的培养物中分离或使用显性选择标记进行选择。这种突变已经被定位并用于创建新的甲病毒载体，用于在哺乳动物细胞中表达非细胞病变基因。这些载体允许以中等水平长期表达，并补充了先前设计用于短期高水平表达的载体。除了用于越来越多的基础研究应用之外，重组甲病毒RNA复制子还可以促进基因疫苗接种和瞬时基因治疗。
Alphaviruses are positive-strand RNA viruses that can mediate efficent cytoplasmic gene expression in insect and vertebrate cells. Through recombinant DNA technology, the alphavirus RNA replication machinery has been engineered for high-level expression of heterologous RNAs and proteins. Amplification of replication-competent alphavirus RNAs (replicons) can be initiated by RNA or DNA transfection and a variety of packaging systems have been developed for producing high titers of infectious viral particles. Although normally cytocidal for vertebrate cells, variants with adaptive mutations allowing noncytopathic replication have been isolated from persistently infected cultures or selected using a dominant selectable marker. Such mutations have been mapped and used to create new alphavirus vectors for noncytopathic gene expression in mammalian cells. These vectors allow long-term expression at moderate levels and complement previous vectors designed for short-term high-level expression. Besides their use for a growing number of basic research applications, recombinant alphavirus RNA replicons may also facilitate genetic vaccination and transient gene therapy.