Introduction: The 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 Method: To 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.Results: Transient 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 VGF, viral 2A sequence and eGFP resulted in eGFP fluorescence in the hypothalamus of both mice and Siberian hamsters, 32 weeks post infusion.
steroid Barreales L steroids, Vazquez Cortes S, Bures P, Ballmer Weber B, Clausen M, Gislason D, Jedrzejczak Czechowicz M, Kowalski ML, Sinaniotis N, Papadopoulos N steroids, Purohit A, de Blay F, Le T M, Knulst AC, Belohlavkova S, Kralimarkova T, Popov T, Asero R steroids, Dubakiene R, Mackie A, Del Prado N, Fernandez Perez C, Mills C, Vieths S, Fernandez Rivas M. 2013. Diagnostic performance of serum specific IgE for milk, egg, peanut, hazelnut, fish steroids, shrimp, celery, apple and peach in the EuroPrevall cross sectional study in allergy clinics. steroid
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