Development and Characterization of "Artificial Viruses" for Gene Delivery in Gene Therapy and DNA Vaccination Studies

• Grant number: 11/15189-6
• Support Opportunities: Scholarships in Brazil - Master
• Start date: June 01, 2012
• End date: August 31, 2013
• Field of knowledge: Engineering - Chemical Engineering
• Principal Investigator: Adriano Rodrigues Azzoni
• Grantee: Rafael Ferraz Alves
• Host Institution: Escola Politécnica (EP). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

A major bottleneck in gene therapy and DNA vaccination protocols using non viral vectors is the low efficiency of gene transfer. This is mainly attributed to the fact that during the traffic to the target cells nuclei, plasmid vectors must overcome a series of physical, enzymatic and diffusional barriers. The main objective of this work is the development of new multifunctional non-viral vectors, also known as "artificial viruses", able to efficiently promote gene delivery to the nucleus of mammalian cells. Binary complexes will be formed by the combination of plasmid DNA (pDNA) and recombinant proteins specifically designed by our group to facilitate pDNA intracellular trafficking. Finally, these complexes will be encapsulated in cationic liposomes formed by the lipids egg fosfatidilcoline (EPC), 1,2-dioleoyl-sn-glycero-3-phosphoetanolamina (DOPE) and 1,2-dioleoyl-3-trimetilamonio-propano (DOTAP) resulting in pseudo-ternary complexes. The development of recombinant transport proteins able to take advantage of strategies used by virus to infect mammalian cells may, in the near future, increase the efficiency of non viral vectors, providing new tools for DNA vaccination and gene therapy studies. The kinetics of formation, stability and physical-chemical parameters such as hydrodynamic diameter and zeta potential will be evaluated and correlated with the efficiency of gene delivery in mammalian cells (HeLa cells). A model pDNA, able to express the reporter gene GFP, will be used in the transfection studies. By this way, we expect to develop new vector able to efficiently transport transgenes and also rise important information about the mechanisms involved in DNA tracking. The development of recombinant transport proteins able to take advantage of strategies used by virus to infect mammalian cells may, in the near future, increase the efficiency of non viral vectors, providing new tools for DNA vaccination and gene therapy studies.

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

ALVES, Rafael Ferraz. Development and characterization of non-viral vectors based in proteins and liposomes to gene delivery.. 2013. Master's Dissertation - Universidade de São Paulo (USP). Escola Politécnica (EP/BC) São Paulo.