ArticleVirology journal2007
Expression of RNA virus proteins by RNA polymerase II dependent expression plasmids is hindered at multiple steps.
Article in Virology journal, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 34 citations in OpenAlex.
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- RSV F evolution escapes some monoclonal antibodies but does not strongly erode neutralization by human polyclonal sera.bioRxiv : the preprint server for biology · 2025Article
- Evaluation of adenoviral vector Ad19a encoding RSV-F as novel vaccine against respiratory syncytial virus.NPJ vaccines · 2024Article
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- Identification of a Human Respiratory Syncytial Virus Cell Entry Inhibitor by Using a Novel Lentiviral Pseudotype System.Journal of virology · 2015Article
- Development of an adenovirus-based respiratory syncytial virus vaccine: preclinical evaluation of efficacy, immunogenicity, and enhanced disease in a cotton rat model.Journal of virology · 2014Article
- Novel vaccine regimen elicits strong airway immune responses and control of respiratory syncytial virus in nonhuman primates.Journal of virology · 2014Article
- Protective efficacy and immunogenicity of a combinatory DNA vaccine against Influenza A Virus and the Respiratory Syncytial Virus.PloS one · 2013Article
- Structure and function of respiratory syncytial virus surface glycoproteins.Current topics in microbiology and immunology · 2013Review
- A respiratory syncytial virus replicon that is noncytotoxic and capable of long-term foreign gene expression.Journal of virology · 2011Article
- Soluble respiratory syncytial virus fusion protein in the fully cleaved, pretriggered state is triggered by exposure to low-molarity buffer.Journal of virology · 2011Article
- Assembly and immunological properties of Newcastle disease virus-like particles containing the respiratory syncytial virus F and G proteins.Journal of virology · 2011Article
- Recombinant respiratory syncytial virus F protein expression is hindered by inefficient nuclear export and mRNA processing.Virus genes · 2010Article
- Refined methods for propagating vesicular stomatitis virus vectors that are defective for G protein expression.Journal of virological methods · 2010Article
- Protective efficacy and immunogenicity of an adenoviral vector vaccine encoding the codon-optimized F protein of respiratory syncytial virus.Journal of virology · 2009Article
- Gene optimization leads to robust expression of human respiratory syncytial virus nucleoprotein and phosphoprotein in human cells and induction of humoral immunity in mice.Journal of virological methods · 2009Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundProteins of human and animal viruses are frequently expressed from RNA polymerase II dependent expression cassettes to study protein function and to develop gene-based vaccines. Initial attempts to express the G protein of vesicular stomatitis virus (VSV) and the F protein of respiratory syncytial virus (RSV) by eukaryotic promoters revealed restrictions at several steps of gene expression.
resultsInsertion of an intron flanked by exonic sequences 5'-terminal to the open reading frames (ORF) of VSV-G and RSV-F led to detectable cytoplasmic mRNA levels of both genes. While the exonic sequences were sufficient to stabilise the VSV-G mRNA, cytoplasmic mRNA levels of RSV-F were dependent on the presence of a functional intron. Cytoplasmic VSV-G mRNA levels led to readily detectable levels of VSV-G protein, whereas RSV-F protein expression remained undetectable. However, RSV-F expression was observed after mutating two of four consensus sites for polyadenylation present in the RSV-F ORF. Expression levels could be further enhanced by codon optimisation.
conclusionInsufficient cytoplasmic mRNA levels and premature polyadenylation prevent expression of RSV-F by RNA polymerase II dependent expression plasmids. Since RSV replicates in the cytoplasm, the presence of premature polyadenylation sites and elements leading to nuclear instability should not interfere with RSV-F expression during virus replication. The molecular mechanisms responsible for the destabilisation of the RSV-F and VSV-G mRNAs and the different requirements for their rescue by insertion of an intron remain to be defined.
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