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

2022; Multidisciplinary Digital Publishing Institute; Volume: 15; Issue: 1 Linguagem: Inglês

10.3390/v15010132

1999-4915

Mayra Colunga-Saucedo, E.I. Rubio-Hernández, Miguel Angel Coronado-Ipiña, Sergio Rosales-Méndoza, Claudia G. Castillo, Mauricio Comas-García,

Virus-based gene therapy research

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.