ArticleNucleic acids research2019
Cell-free expression of RNA encoded genes using MS2 replicase.
Article in Nucleic acids research, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 13 citations in OpenAlex.
- Membraneless protocell confined by a heat flow.Nature physics · 2025Article
- Design of Novel Synthetic RNA Replicons Based onACS synthetic biology · 2024Article
- Emergence of linkage between cooperative RNA replicators encoding replication and metabolic enzymes through experimental evolution.PLoS genetics · 2023Article
- In vitro characterisation of the MS2 RNA polymerase complex reveals host factors that modulate emesviral replicase activity.Communications biology · 2022Article
- Clinical and experimental bacteriophage studies: Recommendations for possible approaches for standing against SARS-CoV-2.Microbial pathogenesis · 2022Review
- Optimising protein synthesis in cell-free systems, a review.Engineering biology · 2021Article
- Applications of phage-derived RNA-based technologies in synthetic biology.Synthetic and systems biotechnology · 2020Review
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
RNA replicases catalyse transcription and replication of viral RNA genomes. Of particular interest for in vitro studies are phage replicases due to their small number of host factors required for activity and their ability to initiate replication in the absence of any primers. However, the requirements for template recognition by most phage replicases are still only poorly understood. Here, we show that the active replicase of the archetypical RNA phage MS2 can be produced in a recombinant cell-free expression system. We find that the 3' terminal fusion of antisense RNAs with a domain derived from the reverse complement of the wild type MS2 genome generates efficient templates for transcription by the MS2 replicase. The new system enables DNA-independent gene expression both in batch reactions and in microcompartments. Finally, we demonstrate that MS2-based RNA-dependent transcription-translation reactions can be used to control DNA-dependent gene expression by encoding a viral DNA-dependent RNA polymerase on a MS2 RNA template. Our study sheds light on the template requirements of the MS2 replicase and paves the way for new in vitro applications including the design of genetic circuits combining both DNA- and RNA-encoded systems.
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Registered trials
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