Alphavirus-based expression vectors: strategies and applications.
Alphaviruses are positive-strand RNA viruses that can mediate efficient 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 alpha-virus 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.
Frolov
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Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110-1093, USA.
Hoffman
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Prágai
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Dryga
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Huang
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Schlesinger
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Rice
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D000524:Alphavirus / Q000235:genetics* / Q000502:physiology; D000818:Animals; D004742:Enhancer Elements, Genetic; D015967:Gene Expression Regulation, Viral; D017930:Genes, Reporter; D005818:Genetic Engineering; D005822:Genetic Vectors; D007313:Insecta; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012093:Replicon; D014162:Transfection / Q000379:methods*; D014714:Vertebrates; D014771:Virion / Q000235:genetics; D014779:Virus Replication
D011993:Recombinant Fusion Proteins