ArticleBMC biotechnology2013
Optimisation of the foot-and-mouth disease virus 2A co-expression system for biomedical applications.
Article in BMC biotechnology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers.
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Who cites it
38 citing papers in PubMed.
- Ribosome dynamics at the conserved PGP motif governs 2A peptide-bond-skipping efficiency.Nucleic acids research · 2026Article
- Vectored Immunoprophylaxis for Mucosal Immunity: Advances and Challenges Associated with Recombinant Secretory IgA Expression.Vaccines · 2026Review
- High Throughput Characterization of Eukaryotic 2A-Like Peptides Identifies Novel Leucine-Associated Reduction in Protein Abundance.bioRxiv : the preprint server for biology · 2026Article
- Molecular architecture and diversity of StopGo/2A translational recoding.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- RNA elements and their biotechnological applications in plants.The New phytologist · 2025Review
- Guidelines for minimal reporting requirements, design and interpretation of experiments involving the use of eukaryotic dual gene expression reporters (MINDR).Nature structural & molecular biology · 2025Review
- Article
- Picornavirus Evolution: Genomes Encoding Multiple 2AViruses · 2024Article
- Highlighter: An optogenetic system for high-resolution gene expression control in plants.PLoS biology · 2023Article
- Immunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine development.Scientific reports · 2023Article
- Easy efficient HDR-based targeted knock-in inBioengineered · 2022Article
- Advanced Fusion Strategies for the Production of Functionalized Potato Virus X Virions.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Synthetic polycistronic sequences in eukaryotes.Synthetic and systems biotechnology · 2021Review
- Proangiogenic Effect of 2A-Peptide Based Multicistronic Recombinant Constructs Encoding VEGF and FGF2 Growth Factors.International journal of molecular sciences · 2021Article
- Light control of the peptide-loading complex synchronizes antigen translocation and MHC I trafficking.Communications biology · 2021Article
- Mammalian expression of virus-like particles as a proof of principle for next generation polio vaccines.NPJ vaccines · 2021Article
- Purification of Pluripotent Stem Cell-Derived Cardiomyocytes Using CRISPR/Cas9-Mediated Integration of Fluorescent Reporters.Methods in molecular biology (Clifton, N.J.) · 2021Article
- A series of dual-reporter vectors for ratiometric analysis of protein abundance in plants.Plant direct · 2020Article
- Validation of a New Multicistronic Plasmid for the Efficient and Stable Expression of Transgenes in Microalgae.International journal of molecular sciences · 2020Article
- Optimization of a 2A self-cleaving peptide-based multigene expression system for efficient expression of upstream and downstream genes in silkworm.Molecular genetics and genomics : MGG · 2019Article
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3 authors.
Funding
Abstract
backgroundMany biomedical applications require the expression or production of therapeutic hetero-multimeric proteins/protein complexes: in most cases only accomplished by co-ordinated co-expression within the same cell. Foot-and-mouth disease virus 2A (F2A) and '2A-like' sequences are now widely used for this purpose. Since 2A mediates a co-translational 'cleavage' at its own C-terminus, sequences encoding multiple proteins (linked via 2As) can be concatenated into a single ORF: a single transgene. It has been shown that in some cases, however, the cleavage efficiency of shorter versions of F2A may be inhibited by the C-terminus of certain gene sequences immediately upstream of F2A. This paper describes further work to optimise F2A for co-expression strategies.
resultsWe have inserted F2A of various lengths in between GFP and CherryFP 'reporter' proteins (in reciprocal or tandem arrangements). The co-expression of these proteins and cleavage efficiencies of F2As of various lengths were studied by in vitro coupled transcription and translation in rabbit reticulocyte lysates, western blotting of HeLa cell lysates and fluorescence microscopy.
conclusionsOptimal and suboptimal lengths of F2A sequences were identified as a result of detailed 'fine-tuning' of the F2A sequence. Based on our data and the model according to which 2A activity is a product of its interaction with the exit tunnel of the ribosome, we suggest the length of the F2A sequence which is not 'sensitive' to the C-terminus of the upstream protein that can be successfully used for co-expression of two proteins for biomedical applications.
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