Diebold, O.; Zhou, S.; Sharp, C.P.; Tesla, B.; Chook, H.W.; Digard, P.; Gaunt, E.R. Towards the Development of a Minigenome Assay for Species A Rotaviruses. Viruses2024, 16, 1396.
Diebold, O.; Zhou, S.; Sharp, C.P.; Tesla, B.; Chook, H.W.; Digard, P.; Gaunt, E.R. Towards the Development of a Minigenome Assay for Species A Rotaviruses. Viruses 2024, 16, 1396.
Diebold, O.; Zhou, S.; Sharp, C.P.; Tesla, B.; Chook, H.W.; Digard, P.; Gaunt, E.R. Towards the Development of a Minigenome Assay for Species A Rotaviruses. Viruses2024, 16, 1396.
Diebold, O.; Zhou, S.; Sharp, C.P.; Tesla, B.; Chook, H.W.; Digard, P.; Gaunt, E.R. Towards the Development of a Minigenome Assay for Species A Rotaviruses. Viruses 2024, 16, 1396.
Abstract
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 viral RdRp activity to be measured as a correlate of reporter protein signal. Here we report the first development of a plasmid-based minigenome assay for species A rotavirus, using a firefly luciferase reporter gene.
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
