Development of non-viral gene delivery vectors based on the Rp3 Dynein light chain

• Grant number: 13/23780-1
• Support Opportunities: Regular Research Grants
• Start date: March 01, 2014
• End date: February 29, 2016
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Adriano Rodrigues Azzoni
• Grantee: Adriano Rodrigues Azzoni
• Host Institution: Escola Politécnica (EP). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

A major limitation on the development of gene therapy and DNA vaccination protocols is the lack of efficient and safe gene delivery vectors. Although safer, non-viral gene delivery vectors must overcome a series of physical, enzymatic and diffusional barriers in order to reach the target cells nuclei. Following the strategy studied by our group, the objective of this project is the delopment and evaluation, "in vitro" and "in vivo", of multifunctional non-viral gene delivery vectors based on recombinant proteins. Our strategy is based on the use of the recombinant dynein light chain Rp3, taking advantage of the ability of the dynein molecular motors to transport cargos from the perifery to the center of the cells. The Rp3 was fusioned to synthetic peptides, improving its ability to interact and condense DNA (DNA-binding sequence), transverse membranes (TAT membrane active peptide) and escape from endocitic vesicles (histidine sequence). The aim of this work is the improvement of the understanding on the mechanisms involved on the cell uptake and intracellular trafficking of these vectors. By combining different analytical assays such as dynamic light scattering, zeta potencial, interaction assays, and transfections on the presence of especific drugs, we believe will be able to correlate the physical-chemical properties of the vectors with the intracellular trafficking and gene delivery efficiency. The transfection of different cell lines will increase our understanding on the "in vitro" efficiency of these vectors, and the "in vivo" evaluation using a an animal model for cystic fibrosis (mice) will be a further step on the understanding of the use of recombinant protein vectors for gene delivery. (AU)

Scientific publications

(4)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ALVES, RAFAEL F.; FAVARO, MARIANNA T. P.; BALBINO, TIAGO A.; DE TOLEDO, MARCELO A. S.; DE LA TORRE, LUCIMARA G.; AZZONI, ADRIANO R.. Recombinant protein-based nanocarriers and their association with cationic liposomes: Characterization and in vitro evaluation. COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS, v. 513, p. 1-10, JAN 5 2017. (12/23143-9, 13/23780-1)

CARDOSO, SARA; SOUSA, ANGELA; QUEIROZ, JOAO A.; AZZONI, ADRIANO R.; SOUSA, FANI. Arginine homopeptides for plasmid DNA purification using monolithic supports. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, v. 1087, p. 149-157, JUN 15 2018. (13/23780-1)

DE PINHO FAVARO, MARIANNA TEIXEIRA; UNZUETA, UGUTZ; DE CABO, MARTI; VILLAVERDE, ANTONIO; FERRER-MIRALLES, NEUS; AZZONI, ADRIANO RODRIGUES. Intracellular trafficking of a dynein-based nanoparticle designed for gene delivery. European Journal of Pharmaceutical Sciences, v. 112, p. 71-78, JAN 15 2018. (12/18850-8, 13/23780-1)

CARDOSO, SARA; PESSOA FILHO, PEDRO DE ALCANTARA; SOUSA, FANI; AZZONI, ADRIANO RODRIGUES. Arginine and di-arginine ligands for plasmid DNA purification using negative chromatography. Separation and Purification Technology, v. 202, p. 281-289, AUG 31 2018. (13/23780-1)