In vitro evaluation of the gene delivery of rabies virus glycoprotein gene using protein based non-viral vectors

• Grant number: 14/14404-9
• Support Opportunities: Scholarships in Brazil - Master
• Start date: October 01, 2014
• End date: September 30, 2016
• Field of knowledge: Engineering - Chemical Engineering
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Adriano Rodrigues Azzoni
• Grantee: Daniela Flores Teruya Astudillo
• Host Institution: Escola Politécnica (EP). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

A major factor limiting the development and approval for human use of DNA vaccines is the lack of an ideal vector for gene delivery that is both efficient and safe. Although safer, non-viral vectors face a number of physical, enzymatic and diffusional barriers that limit the delivery of transgenes to the nucleus of the target cells. These limitations are even more important when it comes to the delivery of genes to macrophages, interesting from the point of view of development of genetic vaccines. Continuing the work developed by our group, the main objective of this project is to evaluate the performance of multifunctional non-viral vectors based on recombinant proteins in gene delivery to macrophages (IC23 lineage). Our approach uses recombinant protein T-Rp3 a modified light chain dynein, thus seeking to exploit the natural ability of molecular motors to transport loads from the periphery to the interior of the cells via microtubules network. Plasmids (pDNA) model containing the GFP reporter gene or the rabies virus glycoprotein (GPV) will be used in this study. Combining the different techniques such as light scattering, zeta potential and transfection assays in the presence of drugs, we expect to correlate the physicochemical properties of the pDNA-protein complex with the intracellular trafficking and efficiency of gene delivery. The transfection of various mammalian cell types will allow us to evaluate the efficacy of our gene delivery vectors to perform in vitro, especially to macrophages. We hope to increase the currently available knowledge on the use of recombinant proteins as gene delivery vectors with potential for the development of new vaccines against rabies. (AU)

Academic Publications

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

ASTUDILLO, Daniela Flores Teruya. In vitro evaluation of the rables virus glycoprotein gene delivery using non viral vectors.. 2016. Master's Dissertation - Universidade de São Paulo (USP). Escola Politécnica (EP/BC) São Paulo.