Towards the Development of a Minigenome Assay for Species A Rotaviruses

Autor: Ola Diebold, Shu Zhou, Colin Peter Sharp, Blanka Tesla, Hou Wei Chook, Paul Digard, Eleanor R. Gaunt
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: Viruses, Vol 16, Iss 9, p 1396 (2024)
Druh dokumentu: article
ISSN: 16091396
1999-4915
DOI: 10.3390/v16091396
Popis: RNA virus polymerases carry out multiple functions necessary for successful genome replication and transcription. A key tool for molecular studies of viral RNA-dependent RNA polymerases (RdRps) is a ‘minigenome’ or ‘minireplicon’ assay, in which viral RdRps are reconstituted in cells in the absence of full virus infection. Typically, plasmids expressing the viral polymerase protein(s) and other co-factors are co-transfected, along with a plasmid expressing an RNA encoding a fluorescent or luminescent reporter gene flanked by viral untranslated regions containing cis-acting elements required for viral RdRp recognition. This reconstitutes the viral transcription/replication machinery and allows the viral RdRp activity to be measured as a correlate of the reporter protein signal. Here, we report on the development of a ‘first-generation’ plasmid-based minigenome assay for species A rotavirus using a firefly luciferase reporter gene.
Databáze: Directory of Open Access Journals
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