Noncytopathic Sindbis virus RNA vectors for heterologous gene expression
Autor: | Charles M. Rice, Béla M. Prágai, Sondra Schlesinger, Ilya Frolov, Brett D. Lindenbach, Eugene Agapov |
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Rok vydání: | 1998 |
Předmět: |
Sindbis virus
Transcription Genetic viruses Genetic Vectors Green Fluorescent Proteins Kidney Transfection Cell Line Viral Proteins Plasmid Genes Reporter Cricetinae Gene expression Humans RNA Viruses Animals Replicon Cloning Molecular Gene Selectable marker Subgenomic mRNA Multidisciplinary biology DNA-Directed RNA Polymerases Biological Sciences biology.organism_classification beta-Galactosidase Virology Recombinant Proteins Luminescent Proteins Electroporation Viral replication Commentary RNA Viral Sindbis Virus Plasmids |
Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 95(22) |
ISSN: | 0027-8424 |
Popis: | Infection of vertebrate cells with alphaviruses normally leads to prodigious expression of virus-encoded genes and a dramatic inhibition of host protein synthesis. Recombinant Sindbis viruses and replicons have been useful as vectors for high level foreign gene expression, but the cytopathic effects of viral replication have limited their use to transient studies. We recently selected Sindbis replicons capable of persistent, noncytopathic growth in BHK cells and describe here a new generation of Sindbis vectors useful for long-term foreign gene expression based on such replicons. Foreign genes of interest as well as the dominant selectable marker puromycinN-acteyltransferase, which confers resistance to the drug puromycin, were expressed as subgenomic transcripts of noncytopathic replicons or defective-interfering genomes complemented in trans by a replicon. Based on these strategies, we developed vectors that can be initiated via either RNA or DNA transfection and analyzed them for their level and stability of foreign gene expression. Noncytopathic Sindbis vectors express reasonably high levels of protein in nearly every cell. These vectors should prove to be flexible tools for the rapid expression of heterologous genes under conditions in which cellular metabolism is not perturbed, and we illustrate their utility with a number of foreign proteins. |
Databáze: | OpenAIRE |
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