4.6 Article

Construction of a Chikungunya Virus, Replicon, and Helper Plasmids for Transfection of Mammalian Cells

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VIRUSES-BASEL
卷 15, 期 1, 页码 -

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MDPI
DOI: 10.3390/v15010132

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Chikungunya virus; alphavirus DNA vectors; replicon plasmid; helper plasmid

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In this study, three plasmids were generated for the transfection of mammalian cells: an infectious clone of Chikungunya virus (CHIKV), a helper plasmid coding for the structural proteins, and a replicon plasmid coding for nonstructural proteins. All plasmids contain a reporter gene (mKate2). Co-transfection of the helper and replicon plasmids has biotechnological/biomedical applications as it enables the delivery of self-replicating RNA for transient gene expression in target cells.
The genome of Alphaviruses can be modified to produce self-replicating RNAs and virus-like particles, which are useful virological tools. In this work, we generated three plasmids for the transfection of mammalian cells: an infectious clone of Chikungunya virus (CHIKV), one that codes for the structural proteins (helper plasmid), and another one that codes nonstructural proteins (replicon plasmid). All of these plasmids contain a reporter gene (mKate2). The reporter gene in the replicon RNA and the infectious clone are synthesized from subgenomic RNA. Co-transfection with the helper and replicon plasmids has biotechnological/biomedical applications because they allow for the delivery of self-replicating RNA for the transient expression of one or more genes to the target cells.

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