ArticleJournal of neuroscience methods2015
The use of a viral 2A sequence for the simultaneous over-expression of both the vgf gene and enhanced green fluorescent protein (eGFP) in vitro and in vivo.
Article in Journal of neuroscience methods, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- Retrograde transduction of dopaminergic cells in substantia nigra of the rhesus monkey.Scientific reports · 2026Article
- Retrograde transduction of dopaminergic cells in substantia nigra of rhesus monkey.bioRxiv : the preprint server for biology · 2026Article
- Evaluation of the SH-SY5Y cell line as anFrontiers in molecular neuroscience · 2023Article
- Co-delivery of genes can be confounded by bicistronic vector design.MRS communications · 2022Article
- 2A and 2A-like Sequences: Distribution in Different Virus Species and Applications in Biotechnology.Viruses · 2021Review
- The Bacterial Enzyme RfxCas13d Is Less Neurotoxic Than PspCas13b and Could Be a Promising RNA Editing and Interference Tool in the Nervous System.Brain sciences · 2021Article
- Expression of aFrontiers in cellular and infection microbiology · 2020Article
- Adipocytes: A Novel Target for IL-15/IL-15Rα Cancer Gene Therapy.Molecular therapy : the journal of the American Society of Gene Therapy · 2019Article
- Article
- Hypothalamic over-expression of VGF in the Siberian hamster increases energy expenditure and reduces body weight gain.PloS one · 2017Article
- Thyroid hormone and vitamin D regulate VGF expression and promoter activity.Journal of molecular endocrinology · 2016Article
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
introductionThe viral 2A sequence has become an attractive alternative to the traditional internal ribosomal entry site (IRES) for simultaneous over-expression of two genes and in combination with recombinant adeno-associated viruses (rAAV) has been used to manipulate gene expression in vitro. NEW
methodTo develop a rAAV construct in combination with the viral 2A sequence to allow long-term over-expression of the vgf gene and fluorescent marker gene for tracking of the transfected neurones in vivo.
resultsTransient transfection of the AAV plasmid containing the vgf gene, viral 2A sequence and eGFP into SH-SY5Y cells resulted in eGFP fluorescence comparable to a commercially available reporter construct. This increase in fluorescent cells was accompanied by an increase in VGF mRNA expression. Infusion of the rAAV vector containing the vgf gene, viral 2A sequence and eGFP resulted in eGFP fluorescence in the hypothalamus of both mice and Siberian hamsters, 32 weeks post infusion. In situ hybridisation confirmed that the location of VGF mRNA expression in the hypothalamus corresponded to the eGFP pattern of fluorescence. COMPARISON WITH OLD
methodThe viral 2A sequence is much smaller than the traditional IRES and therefore allowed over-expression of the vgf gene with fluorescent tracking without compromising viral capacity.
conclusionThe use of the viral 2A sequence in the AAV plasmid allowed the simultaneous expression of both genes in vitro. When used in combination with rAAV it resulted in long-term over-expression of both genes at equivalent locations in the hypothalamus of both Siberian hamsters and mice, without any adverse effects.
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