Version 1
: Received: 27 April 2024 / Approved: 28 April 2024 / Online: 29 April 2024 (10:24:58 CEST)
How to cite:
Diebold, O.; Sharp, C. P.; Tesla, B.; Chook, H. W.; Digard, P.; Gaunt, E. R. Development of a Mini-Genome Assay for Species a Rotaviruses. Preprints2024, 2024041878. https://doi.org/10.20944/preprints202404.1878.v1
Diebold, O.; Sharp, C. P.; Tesla, B.; Chook, H. W.; Digard, P.; Gaunt, E. R. Development of a Mini-Genome Assay for Species a Rotaviruses. Preprints 2024, 2024041878. https://doi.org/10.20944/preprints202404.1878.v1
Diebold, O.; Sharp, C. P.; Tesla, B.; Chook, H. W.; Digard, P.; Gaunt, E. R. Development of a Mini-Genome Assay for Species a Rotaviruses. Preprints2024, 2024041878. https://doi.org/10.20944/preprints202404.1878.v1
APA Style
Diebold, O., Sharp, C. P., Tesla, B., Chook, H. W., Digard, P., & Gaunt, E. R. (2024). Development of a Mini-Genome Assay for Species a Rotaviruses. Preprints. https://doi.org/10.20944/preprints202404.1878.v1
Chicago/Turabian Style
Diebold, O., Paul Digard and Eleanor R Gaunt. 2024 "Development of a Mini-Genome Assay for Species a Rotaviruses" Preprints. https://doi.org/10.20944/preprints202404.1878.v1
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.
