Enhancement of plasmid-mediated transgene expression.
A large number of studies aimed at the treatment of cancer, autoimmune and metabolic diseases, neurodegenerative disorders, allergic diseases, as well as muscle disorders strengthen the fact that gene therapy could represent an alternative method to treat human diseases where conventional approaches are less effective. To improve transgene expression from plasmid vectors, DNA nuclear targeting sequences (DTSs) can be introduced in a vector backbone to increase in vivo expression up to 20-fold using electroporation (EP) delivery in muscle tissue. The purpose of this chapter is to represent a step-by-step strategy for the construction of a plasmid vector with enhanced efficiency of nuclear plasmid uptake and the methodic for the in vivo efficiency evaluation of the obtained expression vector.
Section of Medical Biotechnology, Institute of Translational Pharmacology (IFT), National Research Council (CNR), via Fosso del Cavaliere 100, 00133, Rome, Italy.
Daniela
D
Fioretti
Sandra
S
Iurescia
Monica
M
Rinaldi
D000818:Animals; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology; D014162:Transfection / Q000379:methods; D019076:Transgenes / Q000235:genetics* / Q000276:immunology; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
D019444:Vaccines, DNA